TWI811982B - 對共刺激tnf受體具特異性之雙特異性抗體 - Google Patents
對共刺激tnf受體具特異性之雙特異性抗體 Download PDFInfo
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Abstract
本發明係關於新穎雙特異性抗原結合分子,其包含(a)至少一個能夠特異性結合於目標細胞抗原之部分體,(b)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,及(c)由能夠穩定結合之第一子單元及第二子單元構成之Fc結構域,且係關於製造此等分子之方法及其使用方法。
Description
本發明係關於新穎雙特異性抗原結合分子,其包含(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一子單元及第二子單元構成之Fc結構域。本發明亦係關於此等分子之製備方法及其使用方法。
若干腫瘤壞死因子受體(TNFR)家族之成員在初始T細胞活化之後用於支持T細胞反應,且因此在免疫系統之組構及功能中起關鍵作用。CD27、4-1BB (CD137)、OX40 (CD134)、HVEM、CD30及GITR可對T細胞具有共刺激作用,意謂其在初始T細胞活化之後支持T細胞反應(Watts T.H. (2005) Annu. Rev. Immunol. 23, 23-68)。此等共刺激TNFR家族成員之作用通常可與CD28之作用及彼此功能上、時間上或空間上分離。 不同共刺激因子對T細胞活化/存活信號之依序及短暫調節可用於允許反應耐久性,同時維持對T細胞存活之密切控制。視疾病病狀而定,共刺激TNF家族成員之刺激可加重或改善疾病。儘管此等複雜性,但TNFR家族共刺激因子之刺激或阻斷展現若干治療性應用(包括癌症、傳染病、移植及自體免疫)之前景。
在若干共刺激分子中,腫瘤壞死因子(TNF)受體家族成員OX40(CD134)在效應子及記憶T細胞之存活及恆定起關鍵作用(Croft M.等人. (2009), Immunological Reviews 229, 173-191)。OX40 (CD134)在若干細胞類型中表現且針對感染、腫瘤及自身抗原調節免疫反應,且已證實其在T細胞、NKT-細胞及NK細胞以及嗜中性白細胞表面上表現(Baumann R.等人. (2004), Eur. J. Immunol. 34, 2268-2275)且顯示回應於多種刺激信號精確誘導或強烈上調。已在每一種OX40表現細胞類型中證實分子之功能活性,表明活體內OX40介導活性之複雜調節。與T細胞受體觸發相組合,T細胞上OX40經其天然配體或促效抗體嚙合導致PI3K及NFκB信號傳導路徑的協同活化(Song J.等人. (2008) J. Immunology 180(11), 7240-7248)。繼而,此導致加強之增殖、增加之細胞激素受體及細胞激素產生以及更佳之活化T細胞存活。除了在效應子CD4
+或CD8
+T細胞中的共刺激之外,OX40觸發最近已顯示抑制衍生自原生CD4
+T細胞的T調節細胞的發育及免疫抑制功能。此作用可能至少部分負責提高OX40對抗腫瘤或抗微生物免疫反應之活性。由於OX40嚙合可擴大T細胞群體,促進細胞激素分泌以及支持T細胞記憶,因此包括配體OX40L之抗體及可溶性形式的促效劑已成功地用於多種臨床前腫瘤模型(Weinberg等人. (2000), J. Immunol. 164, 2160-2169)。
4-1BB (CD137)為TNF受體超家族之成員,已最先鑑別為由T細胞活化誘導表現之分子(Kwon Y.H.及Weissman S.M. (1989), Proc. Natl. Acad. Sci. USA 86, 1963-1967)。後續研究證實4-1BB於T-及B-淋巴細胞(Snell L.M.等人. (2011) Immunol. Rev. 244, 197-217或Zhang X.等人(2010), J. Immunol. 184, 787-795)、NK細胞(Lin W.等人. (2008), Blood 112, 699-707)、NKT細胞(Kim D.H.等人. (2008), J. Immunol. 180, 2062-2068)、單核細胞(Kienzle G.及von Kempis J. (2000), Int. Immunol. 12, 73-82或Schwarz H.等人(1995), Blood 85, 1043-1052)、嗜中性白細胞(Heinisch I.V.等人(2000), Eur. J. Immunol. 30, 3441-3446)、肥大細胞(Nishimoto H.等人. (2005), Blood 106, 4241-4248),及樹突狀細胞以及非造血來源之細胞,諸如內皮細胞及平滑肌細胞(Broll K.等人. (2001), Am. J. Clin. Pathol. 115, 543-549或Olofsson P.S.等人. (2008), Circulation 117, 1292-1301)中之表現。4-1BB於不同細胞類型中之表現大部分可由多種刺激信號誘導及驅動,諸如T細胞受體(TCR)或B細胞受體觸發,以及經共刺激分子或促炎性細胞激素受體誘導之信號傳導(Diehl L.等人. (2002), J. Immunol. 168, 3755-3762;von Kempis J.等人. (1997), Osteoarthritis Cartilage 5, 394-406;Zhang X.等人. (2010), J. Immunol. 184, 787-795)。
已知CD137信號傳導刺激IFNγ分泌及NK細胞增殖(Buechele C.等人. (2012), Eur. J. Immunol. 42, 737-748;Lin W.等人. (2008), Blood 112, 699-707;Melero I.等人. (1998), Cell Immunol. 190, 167-172)以及促進DC活化,由其存活率提高及分泌細胞激素及上調共刺激分子之能力指示(Choi B. K.等人. (2009), J. Immunol. 182, 4107-4115;Futagawa T.等人. (2002), Int. Immunol. 14, 275-286;Wilcox R. A.等人. (2002), J. Immunol. 168, 4262-4267)。然而,CD137最佳表徵為共刺激分子,其調節T細胞之CD4
+及CD8
+子集中TCR誘導之活化。與TCR觸發組合,促效4-1BB特異性抗體提高T細胞之增殖、刺激淋巴激素分泌及降低T-淋巴細胞對活化誘導之細胞死亡的靈敏度(Snell L.M.等人. (2011) Immunol. Rev. 244, 197-217)。符合4-1BB抗體對T細胞之此等活體外共刺激作用,其投與腫瘤攜帶小鼠導致許多實驗腫瘤模型中的強抗腫瘤作用(Melero I.等人. (1997), Nat. Med. 3, 682-685;Narazaki H.等人. (2010), Blood 115, 1941-1948)。活體內消耗實驗表明CD8
+T細胞在4-1BB特異性抗體之抗腫瘤作用中起最重要作用。然而,視包括抗-4-1BB之腫瘤模型或組合療法而定,已報導其它類型之細胞(諸如DC、NK細胞或CD4
+T細胞)(Murillo O.等人. (2009), Eur. J. Immunol. 39, 2424-2436;Stagg J.等人. (2011), Proc. Natl. Acad. Sci. USA 108, 7142-7147)。
除了對不同淋巴細胞亞群的直接作用之外,4-1BB促效劑亦可經腫瘤血管內皮上4-1BB介導之細胞間黏附分子1(ICAM1)及血管細胞黏附分子1(VCAM1)的上調來誘發活化T細胞在腫瘤中之浸潤及滯留(Palazon A.等人. (2011), Cancer Res. 71, 801-811)。4-1BB觸發亦可逆轉暴露於可溶性抗原誘發之T細胞失能的狀態,該可溶性抗原可促成腫瘤微環境中或慢性感染期間免疫耐受性破壞(Wilcox R.A.等人. (2004), Blood 103, 177-184)。
4-1BB促效抗體之活體內免疫調節特性看起來需要抗體分子上存在野生型Fc部分體,由此暗示Fc受體結合為此類試劑之藥理學活性所需的重要事件,如已經針對對TNFR超家族之其他細胞凋亡誘發或免疫調節成員具有特異性之促效抗體所述(Li F.及Ravetch J.V. (2011), Science 333, 1030-1034;Teng M.W.等人. (2009), J. Immunol. 183, 1911-1920)。然而,全身性投與具有功能活性Fc結構域之4-1BB特異性促效抗體亦誘發與肝臟毒性有關的CD8
+T細胞之擴展(Dubrot J.等人. (2010), Cancer Immunol. Immunother. 59, 1223-1233),此在小鼠體內在無功能性Fc受體存在下減弱或顯著改善。在人類臨床試驗(ClinicalTrials.gov, NCT00309023)中,每三週一次投與Fc勝任型4-1BB促效抗體(BMS-663513)持續12週可誘發黑素瘤、卵巢或腎細胞癌之患者中之疾病穩定。然而,另一試驗中之相同抗體(NCT00612664)引起4級肝炎,導致試驗終止(Simeone E.及Ascierto P.A. (2012), J. Immunotoxicology 9, 241-247)。因此,需要新一代促效劑,其應不僅有效嚙合造血及內皮細胞之表面上的4-1BB,且亦能夠經除結合於Fc受體以外的機制實現,以避免不可控制的副作用。
可用臨床前及臨床資料明確證明對共刺激TNFR家族成員(諸如Ox40及4-1BB)之有效促效劑的高度臨床需要,其能夠誘發及提高對癌症的有效內源性免疫反應。然而,該等作用幾乎從不限於單個細胞類型或經單一機制起作用,且設計成說明細胞間及細胞內信號傳導機制之研究已揭示提高的複雜性水準。因此,需要較佳對單一細胞類型起作用的「靶向」促效劑。本發明的抗原結合分子組合能夠較佳結合於腫瘤特定性或腫瘤相關目標之部分體與能夠促效結合於共刺激TNF受體之部分體。本發明的抗原結合分子可能夠不僅有效觸發TNF受體,而且在所要部位極具選擇性,由此減少不合意的副作用。
本發明係關於雙特異性抗原結合分子,其組合至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體(即至少一個靶向共刺激TNF受體之抗原結合位點)與至少一個能夠特異性結合於目標細胞抗原之部分體(即至少一個靶向目標細胞抗原之抗原結合側)。由於對目標細胞抗原之結合能力,此等雙特異性抗原結合分子是有利的,因為其將較佳活化共刺激TNF受體的表現目標細胞抗原之部位。本發明亦提供能夠特異性結合於共刺激TNF受體家族成員之新穎抗體。相較於共刺激TNF受體之已知抗體,此等抗體具有對併入至雙特異性抗原結合分子與能夠特異性結合於目標細胞抗原之部分體的組合有利的特性。
在一個態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a)至少一個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b)至少一個能夠特異性結合於目標細胞抗原之部分體,
及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個特定態樣中,雙特異性抗原結合分子包含(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,其中該共刺激TNF受體家族成員選自由OX40及4-1BB組成之群,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個態樣中,共刺激TNF受體家族成員為OX40。因此,在一個特定態樣中,能夠特異性結合於共刺激TNF受體家族成員之部分體結合於包含SEQ ID NO:1之胺基酸序列的多肽。
在另一態樣中,提供雙特異性抗原結合分子,其包含至少一個能夠特異性結合於OX40之部分體,其中該部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:2及SEQ ID NO:3組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:4及SEQ ID NO:5組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO: 9、SEQ ID NO:10、SEQ ID NO:11及SEQ ID NO:12組成之群的胺基酸序列,
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:13、SEQ ID NO:14及SEQ ID NO:15組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:16、SEQ ID NO:17及SEQ ID NO:18組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23及SEQ ID NO:24組成之群的胺基酸序列。
在另一態樣中,本發明提供一種雙特異性抗原結合分子,其中能夠特異性結合於OX40之部分體包含重鏈可變區VH,其包含與選自由SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO: 29、SEQ ID NO: 31、SEQ ID NO: 33、SEQ ID NO:35及SEQ ID NO: 37組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區VL,其包含與胺基酸序列SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO: 30、SEQ ID NO: 32、SEQ ID NO: 34、SEQ ID NO:36及SEQ ID NO: 38至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其中能夠特異性結合於OX40之部分體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
在另一態樣中,共刺激TNF受體家族成員為4-1BB。因此,在一個特定態樣中,能夠特異性結合於共刺激TNF受體家族成員之部分體結合於包含SEQ ID NO:39之胺基酸序列的多肽。
此外,提供雙特異性抗原結合分子,其包含至少一個能夠特異性結合於4-1BB之部分體,其中該部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:40及SEQ ID NO:41組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:42及SEQ ID NO:43組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47及SEQ ID NO:48組成之群的胺基酸序列
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:49及SEQ ID NO:50組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:51及SEQ ID NO:52組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:53、SEQ ID NO:54、SEQ ID NO:55、SEQ ID NO:56及SEQ ID NO:57組成之群的胺基酸序列。
在另一態樣中,本發明提供一種雙特異性抗原結合分子,其中能夠特異性結合於4-1BB之部分體包含重鏈可變區VH,其包含與選自由SEQ ID NO:58、SEQ ID NO:60、SEQ ID NO:62、SEQ ID NO:64及SEQ ID NO:66組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與選自由SEQ ID NO:59、SEQ ID NO:61、SEQ ID NO:63、SEQ ID NO:65及SEQ ID NO:67組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其中能夠特異性結合於4-1BB之部分體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 至少一個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 至少一個能夠特異性結合於目標細胞抗原之部分體,
及
(c) 由能夠穩定結合之第一子單元及第二子單元構成之Fc結構域,其中能夠特異性結合於共刺激TNF受體家族成員之部分體為Fab片段。在一個態樣中,若雙特異性抗原結合分子包含超過一個能夠特異性結合於共刺激TNF受體家族成員之部分體,則所有能夠特異性結合於共刺激TNF受體家族成員之部分體為Fab片段。
在另一態樣中,雙特異性抗原結合分子包含(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,其中該目標細胞抗原選自由以下組成之群:纖維母細胞活化蛋白質(FAP)、黑素瘤相關之硫酸軟骨素蛋白聚糖(MCSP)、表皮生長因子受體(EGFR)、癌胚抗原(CEA)、CD19、CD20及CD33,及(c)由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。更特定言之,目標細胞抗原為纖維母細胞活化蛋白質(FAP)。
在一個特定態樣中,提供雙特異性抗原結合分子,其中能夠特異性結合於FAP之部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:68及SEQ ID NO:69組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:70及SEQ ID NO:71組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:72及SEQ ID NO:73組成之群的胺基酸序列,
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:74及SEQ ID NO:75組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:76及SEQ ID NO:77組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:78及SEQ ID NO:79組成之群的胺基酸序列。
在另一態樣中,本發明因此提供一種雙特異性抗原結合分子,其中
(i) 能夠特異性結合於OX40之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:25、SEQ ID NO: 27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35或SEQ ID NO:37至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:26、SEQ ID NO: 28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQ ID NO:36或SEQ ID NO:38至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及
(ii) 能夠特異性結合於FAP之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:80或SEQ ID NO:82至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:81或SEQ ID NO:83至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
在一個態樣中,本發明提供一種雙特異性抗原結合分子,其中
(i) 能夠特異性結合於OX40之部分體包含包括胺基酸序列SEQ ID NO:27的重鏈可變區VH及包括胺基酸序列SEQ ID NO:28的輕鏈可變區,及
(ii) 能夠特異性結合於FAP之部分體包含包括胺基酸序列SEQ ID NO:82的重鏈可變區VH及包括胺基酸序列SEQ ID NO:83的輕鏈可變區。在另一態樣中,本發明提供一種雙特異性抗原結合分子,其中
(i) 能夠特異性結合於4-1BB之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:58、SEQ ID NO: 60、SEQ ID NO:62、SEQ ID NO:64或SEQ ID NO:66至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:59、SEQ ID NO: 61、SEQ ID NO:63、SEQ ID NO:65或SEQ ID NO:67至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及
(ii) 能夠特異性結合於FAP之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:80或SEQ ID NO:82至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:81或SEQ ID NO:83至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
在另一態樣中,提供雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,
(b) 能夠特異性結合於目標細胞抗原之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個態樣中,雙特異性抗原結合分子為人類、人類化或嵌合抗體。特定言之,Fc結構域為人類IgG結構域,詳言之IgG1 Fc結構域或IgG4 Fc結構域。
在另一態樣中,提供一種雙特異性抗原結合分子,其中Fc結構域包含一或多個降低抗體對Fc受體之結合親和力及/或效應功能的胺基酸取代。特定言之,Fc結構域為具有胺基酸突變L234A、L235A及P329G(根據Kabat EU索引編號)的人類IgG1子類。
在另一態樣中,提供一種雙特異性抗原結合分子,其中該Fc結構域包含促進Fc結構域之第一子單元及第二子單元結合的修飾。在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其中根據杵-臼方法,Fc結構域之第一子單元包含杵且Fc結構域之第二子單元包含臼。更特定言之,Fc結構域之第一子單元包含胺基酸取代S354C及T366W(根據Kabat EU索引編號)且Fc結構域之第二子單元包含胺基酸取代Y349C、T366S及Y407V(根據Kabat EU索引之編號)。
特定言之,本發明提供一種雙特異性抗原結合分子,其包含
(a) 兩個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 兩個能夠特異性結合於目標細胞抗原之部分體,
及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
因此,提供一種雙特異性抗原結合分子,其中該雙特異性抗原結合分子對於共刺激TNF受體家族成員及目標細胞抗原皆為二價。
在一個特定態樣中,雙特異性抗原結合分子包含
(a) 包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,及
(b) 兩個能夠特異性結合於目標細胞抗原之額外Fab片段,其中該等額外Fab片段各自經肽連接子連接至(a)之重鏈的C端。更特定言之,兩個能夠特異性結合於目標細胞抗原之額外Fab片段為交叉Fab片段,其中可變域VL及VH彼此置換且VL-CH鏈各自經肽連接子連接至(a)之重鏈的C端。
在一個態樣中,兩個能夠特異性結合於共刺激TNF受體家族成員之Fab片段為兩個能夠特異性結合於OX40或4-1BB之Fab片段且兩個能夠特異性結合於目標細胞抗原之額外Fab片段為能夠特異性結合於FAP之交叉Fab片段。
在另一態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a)兩個能夠特異性結合於共刺激TNF受體家族成員之部分體,
(b)一個能夠特異性結合於目標細胞抗原之部分體,
及
(c)由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
因此,提供一種雙特異性抗原結合分子,其中該雙特異性抗原結合分子對於共刺激TNF受體家族成員為二價且對於目標細胞抗原為單價。
在一個特定態樣中,雙特異性抗原結合分子包含
(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域之抗體的兩個輕鏈及兩個重鏈,及
(b) 能夠特異性結合於目標細胞抗原之VH及VL結構域,其中VH結構域經肽連接子連接至重鏈中之一者的C端且其中VL結構域經肽連接子連接至第二重鏈之C端。
在另一態樣中,本發明提供一種特異性結合於OX40之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
在另一態樣中,提供一種特異性結合於4-1BB之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
根據本發明之另一態樣,提供編碼如上文所述之雙特異性抗原結合分子之經分離聚核苷酸或特異性結合於如上文所述之OX40的抗體或特異性結合於如上文所述之4-1BB的抗體。本發明進一步提供載體,特定言之表現載體,其包含本發明之經分離之聚核苷酸,及宿主細胞,其包含本發明之經分離之聚核苷酸或載體。在一些態樣中,宿主細胞為真核細胞,詳言之哺乳動物細胞。
在另一態樣中,提供一種製造如上文所述之雙特異性抗原結合分子或如上文所述之特異性結合於OX40之抗體或如上文所述特異性結合於4-1BB之抗體的方法,其包含步驟(i)在適於表現抗原結合分子之條件下培養本發明的宿主細胞,及(ii)回收抗原結合分子。本發明亦涵蓋藉由本發明方法製備的雙特異性抗原結合分子或特異性結合於OX40之抗體或特異性結合於4-1BB之抗體。
本發明進一步提供一種醫藥組合物,其包含如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如上文所述的特異性結合於4-1BB之抗體及至少一種醫藥學上可接受之賦形劑。
本發明亦涵蓋如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如上文所述的特異性結合於4-1BB之抗體,或包含雙特異性抗原結合分子或特異性結合於OX40之抗體或特異性結合於4-1BB之抗體的醫藥組合物,其用作藥劑。
在一個態樣中,提供如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如上文所述的特異性結合於4-1BB之抗體或本發明之醫藥組合物,其用於
(i) 刺激T細胞反應,
(ii) 支持活化T細胞之存活,
(iii) 治療感染,
(iv) 治療癌症,
(v) 延遲癌症進展,或
(vi) 延長罹患癌症之患者的存活期。
在一個具體實施例中,提供如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如上文所述的特異性結合於4-1BB之抗體或本發明之醫藥組合物,其用於治療癌症。
在另一特定態樣中,本發明提供用於治療癌症的如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如本文所述的特異性結合於4-1BB之抗體,其中雙特異性抗原結合分子與化學治療劑、輻射及/或用於癌症免疫療法的其他試劑組合投與。
在另一態樣中,本發明提供一種抑制個別中腫瘤細胞生長之方法,其包含向該個別投與有效量的如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如上文所述的特異性結合於4-1BB之抗體,或本發明之醫藥組合物來抑制腫瘤細胞生長。
亦提供如上文所述之雙特異性抗原結合分子或如上文所述的特異性結合於OX40之抗體或如上文所述的特異性結合於4-1BB之抗體的用途,其用於製造供治療有需要之個體的疾病的藥劑,特定言之用於製造供治療癌症之藥劑,以及治療個體疾病之方法,其包含向該個別投與治療有效量之組合物,該組合物包含醫藥學上可接受之形式的含有TNF家族配位體三聚物的本發明的抗原結合分子。在一個特定態樣中,疾病為癌症。在上述態樣中之任一者中,個別為哺乳動物,詳言之人類。
定義除非另外定義,否則本文所使用之技術及科學術語具有與本發明所屬領域中通常所使用相同之含義。出於解釋本說明書之目的,將應用以下定義且只要適合,以單數形式使用之術語亦將包括複數且反之亦然。
如本文所用,術語「
抗原結合分子」在其最廣泛的意義上係指特異性結合抗原決定子的分子。抗原結合分子之實例為抗體、抗體片段及骨架抗原結合蛋白。
如本文中所使用,術語「
能夠特異性結合於目標細胞抗原之部分體」係指特異性結合於抗原決定子之多肽分子。在一個態樣中,抗原結合部分能夠經其目標細胞抗原活化信號傳導。在一個特定態樣中,抗原結合部分能夠將其所連接之實體(例如TNF家族配位體三聚體)引導至目標位點,例如引導至特定類型之攜帶抗原決定子之腫瘤細胞或腫瘤基質。能夠特異性結合於目標細胞抗原之部分體包括抗體及其片段,如本文中進一步定義。此外,能夠特異性結合於目標細胞抗原之部分體包括如本文中進一步定義之骨架抗原結合蛋白,例如結合域,其係基於經設計之重複蛋白質或經設計之重複結構域(參看例如WO 2002/020565)。
關於抗體或其片段,術語「能夠特異性結合於目標細胞抗原之部分體」係指包含特異性結合於一部分或所有抗原且與其互補之區域的分子之一部分。可例如藉由一或多個抗體可變域(亦稱為抗體可變區)提供能夠進行特異性抗原結合之部分體。特定言之,能夠進行特異性抗原結合之部分體包含抗體輕鏈可變區(VL)及抗體重鏈可變區(VH)。在一個特定態樣中,「能夠特異性結合於目標細胞抗原之部分體」為Fab片段或交叉Fab片段。
術語「
能夠特異性結合於共刺激 TNF 受體家族成員之部分體」係指特異性結合於共刺激TNF受體家族成員之多肽分子。在一個態樣中,抗原結合部分能夠經共刺激TNF受體家族成員活化信號傳導。能夠特異性結合於目標細胞抗原之部分體包括抗體及其片段,如本文中進一步定義。此外,能夠特異性結合於共刺激TNF受體家族成員之部分體包括如本文進一步定義之骨架抗原結合蛋白,例如基於所設計之重複蛋白或所設計之重複結構域的結合域(參看例如WO 2002/020565)。詳言之,能夠特異性結合於共刺激TNF受體家族成員之部分體包含抗體輕鏈可變區(VL)及抗體重鏈可變區(VH)。在一個特定態樣中,「能夠特異性結合於共刺激TNF受體家族成員之部分體」為Fab片段或交叉Fab片段。
術語「
抗體」在本文中以最廣泛意義使用且涵蓋各種抗體結構,包括(但不限於)單株抗體、多株抗體、單特異性及多特異性抗體(例如雙特異性抗體)及抗體片段,只要其呈現所需抗原結合活性即可。
如本文所用之術語「
單株抗體」係指自實質上均質抗體之群體獲得之抗體,亦即除可能變異抗體(例如含有天然存在之突變或在產生單株抗體製劑期間出現之變異抗體,此類變異體通常以較小量存在)以外,構成該群體之個別抗體相同及/或結合相同抗原決定基。相比於典型地包括針對不同決定子(抗原決定基)之不同抗體的多株抗體製劑,單株抗體製劑之各單株抗體係針對抗原上之單一決定子。
如本文所使用之術語「
單特異性」抗體表示具有一或多個結合位點之抗體,該一或多個結合位點中之每一者結合於相同抗原之相同抗原決定基。術語「
雙特異性」意謂抗原結合分子能夠特異性結合至至少兩個不同的抗原決定子。典型地,雙特異性抗原結合分子包含兩個抗原結合位點,其中之每一者特異性針對不同抗原性決定子。在某些實施例中,雙特異性抗原結合分子能夠同時結合兩個抗原性決定子,詳言之,兩個不同細胞上所表現的兩個抗原性決定子。
如本申請案中所用的術語「
化合價」指示存在對抗原結合分子中一個獨特抗原決定子特異的指定數目的結合位點,其對一個獨特抗原決定子特異。因此,術語「二價」、「四價」及「六價」分別表示抗原結合分子中存在對某一抗原決定子特異的兩個結合位點、四個結合位點及六個結合位點。在本發明之特定態樣中,根據本發明的雙特異性抗原結合分子可以針對某一抗原決定子為單價的,意謂其僅具有一個針對該抗原決定子之結合位點,或其可以針對某一抗原決定子為二價的,意謂其具有兩個針對該抗原決定子之結合位點。
術語「全長抗體」、「完整抗體」及「完全抗體」在本文中可互換使用,其係指具有與原生抗體結構實質上類似之結構的抗體。「
原生抗體」係指具有不同結構之天然存在的免疫球蛋白分子。舉例而言,原生IgG類抗體為約150,000道爾頓(dalton)之雜四聚體糖蛋白,其由二硫鍵鍵結之兩個輕鏈及兩個重鏈構成。自N端至C端,各重鏈具有可變區(VH),亦稱為可變重鏈結構域或重鏈可變域;繼之以三個恆定域(CH1、CH2及CH3),亦稱為重鏈恆定區。類似地,自N端至C端,各輕鏈具有可變區(VL),亦稱為可變輕鏈域或輕鏈可變域,繼之為輕鏈恆定域(CL),亦稱為輕鏈恆定區。抗體之重鏈可歸為五種類型中之一種,該五種類型稱為α (IgA)、δ (IgD)、ε (IgE)、γ (IgG)或μ (IgM),其中一些可進一步分成亞型,例如γ
1(IgG
1)、γ
2(IgG
2)、γ
3(IgG
3)、γ
4(IgG
4)、α
1(IgA
1)及α
2(IgA
2)。抗體輕鏈可基於其恆定域之胺基酸序列而歸為兩種類型之一,稱為卡帕(kappa) (κ)及拉姆達(lambda) (λ)。
「
抗體片段」係指不同於完整抗體,包含完整抗體之一部分且結合完整抗體所結合之抗原的分子。抗體片段之實例包括(但不限於)Fv、Fab、Fab'、Fab'-SH、F(ab')
2;雙功能抗體、三功能抗體、四功能抗體、交叉Fab片段;線性抗體;單鏈抗體分子(例如scFv);及單結構域抗體。關於某些抗體片段之綜述,參看Hudson等人, Nat Med 9, 129-134 (2003)。關於scFv片段之綜述,參看例如Plückthun, The Pharmacology of Monoclonal Antibodies, 第113卷, Rosenburg及Moore編, Springer-Verlag, New York, 第269-315頁 (1994);亦參看WO 93/16185;及美國專利第5,571,894號及第5,587,458號。關於包含救助受體結合抗原決定基殘基及具有延長之活體內半衰期之Fab及F(ab')
2片段的論述,參看美國專利第5,869,046號。雙功能抗體為具有兩個抗原結合位點之抗體片段,其可為二價或雙特異性,參看例如EP 404,097;WO 1993/01161;Hudson等人, Nat Med 9, 129-134 (2003);及Hollinger等人, Proc Natl Acad Sci USA 90, 6444-6448 (1993)。三功能抗體及四功能抗體亦描述於Hudson等人, Nat Med 9, 129-134 (2003)中。單結構域抗體為包含抗體之重鏈可變域的全部或一部分或輕鏈可變域的全部或一部分的抗體片段。在某些實施例中,單結構域抗體為人類單結構域抗體(Domantis, Inc., Waltham, MA;參看例如美國專利第6,248,516 B1號)。抗體片段可藉由多種技術製得,包括(但不限於)完整抗體之蛋白分解消化以及藉由重組型宿主細胞(例如大腸桿菌或噬菌體)產生,如本文中所描述。
完整抗體之番木瓜蛋白酶消化產生兩個相同的抗原結合片段,稱為「Fab」片段,其各含有重鏈及輕鏈可變結構域以及輕鏈之恆定域及重鏈之第一恆定域(CH1)。如本文所用,因此,術語「
Fab 片段」係指包含有包含輕鏈之VL結構域及恆定域(CL)之輕鏈片段,及重鏈之VH結構域及第一恆定域(CH1)的抗體片段。Fab'片段與Fab片段之不同之處在於,在包括一或多個來自抗體鉸鏈區之半胱胺酸之重鏈CH1結構域的羧基端處添加幾個殘基。Fab'-SH為其中恆定域之半胱胺酸殘基具有游離硫醇基之Fab'片段。胃蛋白酶處理產生F(ab')
2片段,其具有兩個抗原組合位點(兩個Fab片段)及Fc區之一部分。根據本發明,術語「Fab片段」亦包括如下文所定義之「交叉Fab片段」或「交換Fab片段」。
術語「
交叉 Fab 片段」或「xFab片段」或「交換Fab片段」係指其中重鏈及輕鏈之可變區或恆定區經互換之Fab片段。交叉Fab分子之兩種不同鏈組成是可能的且包含於本發明之雙特異性抗體中:一方面,Fab重鏈及輕鏈之可變區經互換,即交換Fab分子包含由輕鏈可變區(VL)及重鏈恆定區(CH1)構成之肽鏈以及由重鏈可變區(VH)及輕鏈恆定區(CL)構成之肽鏈。此交換Fab分子亦稱為CrossFab
(VLVH)。另一方面,當Fab重鏈及輕鏈之恆定區互換時,交換Fab分子包含由重鏈可變區(VH)及輕鏈恆定區(CL)構成之肽鏈以及由輕鏈可變區(VL)及重鏈恆定區(CH1)構成之肽鏈。此交換Fab分子亦稱為CrossFab
(CLCH1)。
「單鏈Fab片段」或「
scFab」為由抗體重鏈可變域(VH)、抗體恆定域1 (CH1)、抗體輕鏈可變域(VL)、抗體輕鏈恆定域(CL)及連接子組成之多肽,其中該等抗體結構域及該連接子按N端至C端方向之次序具有以下中之一者:a) VH-CH1-連接子-VL-CL,b) VL-CL-連接子-VH-CH1,c) VH-CL-連接子-VL-CH1或d) VL-CH1-連接子-VH-CL;且其中該連接子為至少30個胺基酸,較佳32與50個胺基酸之間的多肽。該等單鏈Fab片段經由CL結構域與CH1結構域之間的天然雙硫鍵穩定化。此外,此等單鏈Fab分子可經由插入半胱胺酸殘基(例如可變重鏈中之位置44及可變輕鏈中之位置100,根據Kabat編號)而產生鏈間二硫鍵來進一步穩定化。
「交換單鏈Fab片段」或「
x-scFab」為由抗體重鏈可變域(VH)、抗體恆定域1 (CH1)、抗體輕鏈可變域(VL)、抗體輕鏈恆定域(CL)及連接子組成之多肽,其中該等抗體結構域及該連接子按N端至C端方向之次序具有以下中之一者:a) VH-CL-連接子-VL-CH1及b) VL-CH1-連接子-VH-CL;其中VH及VL共同形成抗原結合位點,其特異性結合於抗原且其中該連接子為至少30個胺基酸之多肽。此外,此等x-scFab分子可經由插入半胱胺酸殘基(例如可變重鏈中之位置44及可變輕鏈中之位置100,根據Kabat編號)而產生鏈間二硫鍵來進一步穩定化。
「
單鏈可變片段 (scFv)」為抗體之重鏈(VH)及輕鏈(VL)之可變區之融合蛋白質,其與十至約25個胺基酸之短連接子肽連接。連接子通常富含甘胺酸以具有可撓性,以及絲胺酸或蘇胺酸以具有可溶性,且可連接VH之N端與VL之C端,或反之亦然。儘管移除恆定區且引入連接子,但此蛋白質保留初始抗體之特異性。scFv抗體例如描述於Houston, J.S., Methods in Enzymol. 203 (1991) 46-96)中。此外,抗體片段包含單鏈多肽,其具有VH結構域(亦即能夠與VL結構域一起組裝至功能性抗原結合位點)或VL結構域(亦即能夠與VH結構域一起組裝至功能性抗原結合位點)之特徵,且藉此提供全長抗體之抗原結合性質。
「
骨架抗原結合蛋白」為此項技術中已知,例如纖維結合蛋白及經設計之錨蛋白重複蛋白質(DARPins)已用作抗原結合域之替代性骨架,參看例如Gebauer及Skerra, Engineered protein scaffolds as next-generation antibody therapeutics. Curr Opin Chem Biol 13:245-255 (2009)及Stumpp等人, Darpins: A new generation of protein therapeutics. Drug Discovery Today 13: 695-701 (2008)。在本發明之一個態樣中,骨架抗原結合蛋白係選自由以下組成之群:CTLA-4 ((艾維伯迪)Evibody);脂質運載蛋白(抗運載蛋白);蛋白質A衍生之分子,諸如蛋白質A之Z結構域(親和抗體);A結構域(高親和性多聚體/最大抗體);血清運鐵蛋白(反式體);經設計之錨蛋白重複蛋白質(DARPin);抗體輕鏈或重鏈之可變結構域(單域抗體,sdAb);抗體重鏈之可變結構域(奈米抗體,aVH);V
NAR片段;纖維結合蛋白(阿耐克汀(AdNectin));C型凝集素結構域(四連接素);新型抗原受體β-內醯胺酶之可變結構域(V
NAR片段);人類γ-晶狀體球蛋白或泛素(阿菲林(Affilin)分子);人類蛋白酶抑制劑之kunitz型結構域,微體,諸如來自knottin家族之蛋白質、肽適體及纖維結合蛋白(阿耐克汀)。CTLA-4 (細胞毒性T淋巴細胞相關抗原4)為表現於大部分CD4
+T細胞上之CD28家族受體。其胞外域具有可變結構域類Ig摺疊。對應於抗體之CDR之環可經異源序列取代以賦予不同結合特性。經工程改造以具有不同結合特異性之CTLA-4分子亦稱為艾維伯迪(Evibodies)(例如US7166697B1)。艾維伯迪與抗體(例如結構域抗體)之經分離之可變區之尺寸大致相同。關於其他細節,參看Journal of Immunological Methods 248 (1-2), 31-45 (2001)。脂質運載蛋白為細胞外蛋白質之家族,其傳遞小型疏水性分子,諸如類固醇、後色膽素、類視黃素及脂質。其具有剛性β-片狀第二結構,其在圓錐結構之開放端具有許多環,其可經工程改造以結合於不同目標抗原。抗運載蛋白之尺寸在160-180個胺基酸之間,且來源於脂質運載蛋白。關於其他細節,參看Biochim Biophys Acta 1482: 337-350 (2000)、US7250297B1及US20070224633。親和抗體為來源於金黃色葡萄球菌(Staphylococcus aureus)之蛋白質A之骨架,其可經工程改造以結合於抗原。結構域由具有約58個胺基酸之三螺旋束組成。已藉由表面殘基之隨機化產生文庫。關於其他細節,參看Protein Eng. Des. Sel. 2004, 17, 455-462及EP 1641818A1。高親合性多聚體為來源於A-結構域骨架家族之多域蛋白質。約35個胺基酸之原生結構域採用既定二硫鍵鍵結之結構。藉由改組由A-結構域之家族呈現之天然變化來產生多樣性。關於其他細節,參看Nature Biotechnology 23(12), 1556 - 1561 (2005)及Expert Opinion on Investigational Drugs 16(6), 909-917 (2007年6月)。運鐵蛋白為單體血清傳遞糖蛋白。運鐵蛋白可藉由在允許的表面環中插入肽序列而經工程改造以結合不同目標抗原。經工程改造之運鐵蛋白骨架之實例包括反式體。關於其他細節,參看J. Biol. Chem 274, 24066-24073 (1999)。經設計之錨蛋白重複蛋白質(DARPins)來源於錨蛋白,其為介導整合膜蛋白質與細胞骨架之連接的蛋白質之家族。單一錨蛋白重複為由兩個α螺旋及β迴旋(beta-turn)組成之33殘基主結構。其可藉由隨機化各重複之第一個α螺旋及β迴旋中之殘基而經工程改造以結合不同目標抗原。可藉由增加模組數目來增加其結合界面(親和力成熟方法)。關於其他細節,參看J. Mol. Biol. 332, 489-503 (2003), PNAS 100(4), 1700-1705 (2003)及J. Mol. Biol. 369, 1015-1028 (2007)以及US20040132028A1。單域抗體為由單一單體可變抗體結構域組成之抗體片段。第一個單結構域來源於來自駱駝之抗體重鏈之可變結構域(奈米抗體或V
HH片段)。此外,術語單域抗體包括自主人類重鏈可變域(aVH)或來源於鯊魚之V
NAR片段。纖維結合蛋白為可經工程改造以結合於抗原之骨架。纖連蛋白由III型人類纖維結合蛋白(FN3)之15個重複單元之第10個結構域之天然胺基酸序列之主鏈組成。β-夾層結構之一端處的三個環可經工程改造以使阿耐克汀能夠特異性識別相關治療目標。關於其他細節,參看Protein Eng. Des. Sel. 18, 435- 444 (2005)、US20080139791、WO2005056764及US6818418B1。肽適體為組合性識別分子,其由恆定骨架蛋白質(通常為硫氧還蛋白(TrxA))組成,該恆定骨架蛋白質含有在活性位點處插入之限制性可變肽環。關於其他細節,參看Expert Opin. Biol. Ther. 5, 783-797 (2005)。微體來源於天然存在之長度為25-50個胺基酸之微型蛋白質,其含有3-4個半胱胺酸橋,微型蛋白質之實例包括KalataBI及芋螺毒素(conotoxin)及打結素。微型蛋白質具有環,其可經工程改造以包括至多25個胺基酸而不影響微型蛋白質之整體摺疊。關於經工程改造之打結素結構域之其他細節,參看WO2008098796。
與參考分子「
結合於相同抗原決定基之抗原結合分子」係指一種抗原結合分子,其在競爭分析中阻斷參考分子與其抗原之結合達50%或50%以上,且相反,參考分子在競爭分析中阻斷抗原結合分子與其抗原之結合達50%或50%以上。
術語「
抗原結合域」或「抗原結合位點」係指包含特異性結合於部分或全部抗原且與部分或全部抗原互補之區域的抗原結合分子的部分。當抗原較大時,抗原結合分子可僅結合於抗原之特定一部分,該部分體稱為抗原決定基。抗原結合域可由例如一或多個可變結構域(亦稱為可變區)提供。較佳地,抗原結合域包含抗體輕鏈可變區(VL)及抗體重鏈可變區(VH)。
如本文所用,術語「
抗原決定子」與「抗原」及「抗原決定基」同義且係指多肽大分子上與抗原結合部分結合,形成抗原結合部分-抗原複合物的位點(例如胺基酸之相鄰延伸或由非相鄰胺基酸之不同區域組成的構形組態)。適用的抗原決定子可發現於例如腫瘤細胞表面上、病毒所感染細胞之表面上、其他病變細胞表面上、免疫細胞表面上、游離於血清中及/或細胞外基質(ECM)中。除非另有指示,否則本文中適用作抗原之蛋白質可為來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)及嚙齒動物(例如小鼠及大鼠))之蛋白質的任何原生形式。在一個特定實施例中,抗原為人類蛋白質。在本文中提及特定蛋白質的情況下,該術語涵蓋「全長」的未經處理之蛋白質,以及由細胞中之處理所產生的任何形式之蛋白質。該術語亦涵蓋天然存在之蛋白質變異體,例如剪接變異體或對偶基因變異體。
「
特異性結合」意謂結合對於抗原而言具有選擇性且可與非所需或非特異性相互作用區分。抗原結合分子結合於特異性抗原之能力可經由酶聯結免疫吸附分析(ELISA)或熟習此項技術者熟悉之其他技術(例如表面電漿子共振(SPR)技術(在BIAcore儀器上分析)(Liljeblad et al., Glyco J 17, 323-329 (2000))及傳統結合分析(Heeley, Endocr Res 28, 217-229 (2002))量測。在一個實施例中,抗原結合分子與無關蛋白質之結合程度小於抗原結合分子與抗原之結合的約10%,如藉由例如SPR所量測。在某些實施例中,結合於抗原之分子具有≤1 μM、≤100 nM、≤10 nM、≤1 nM、≤0.1 nM、≤0.01 nM或≤0.001 nM (例如10
- 8M或更低,例如10
- 8M至10
- 13M,例如10
- 9M至10
- 13M)之解離常數(Kd)。
「
親和力」或「結合親和力」係指分子(例如抗體)之單一結合位點與其結合搭配物(例如抗原)之間的非共價相互作用之總和之強度。除非另外指明,否則如本文所使用之「結合親和力」係指反映結合對(例如抗體與抗原)成員之間的1:1相互作用之固有結合親和力。分子X對其搭配物Y之親和力通常可由解離常數(Kd)表示,解離常數為解離速率常數與結合速率常數(分別為koff及kon)之比率。因此,等效親和力可包含不同速率常數,只要速率常數之比率保持相同即可。可藉由此項技術中已知之常用方法(包括本文所描述之彼等方法)來量測親和力。一種用於量測親和力之具體方法為表面電漿子共振(SPR)。
「
親和力成熟」抗體係指相較於在一或多個高變區(HVR)中不具有一或多個變化之親本抗體,具有此類變化之抗體,此等變化使抗體對抗原之親和力得到改良。
如本文所用,「
目標細胞抗原」係指目標細胞(例如腫瘤細胞,諸如癌細胞或腫瘤基質細胞)表面上所呈遞的抗原決定子。在某些實施例中,目標細胞抗原為腫瘤細胞之表面上的抗原。在一個實施例中,目標細胞抗原係選自由以下組成之群:纖維母細胞活化蛋白質(FAP)、癌胚抗原(CEA)、黑素瘤相關軟骨素硫酸鹽蛋白聚糖(MCSP)、表皮生長因子受體(EGFR)、CD19、CD20及CD33。特定言之,目標細胞抗原為纖維母細胞活化蛋白質(FAP)。
除非另有指示,否則術語「
纖維母細胞活化蛋白質 ( FAP )」,亦稱為脯胺醯基內肽酶FAP或Seprase (EC 3.4.21),係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生FAP。該術語涵蓋「全長」的未經處理之FAP以及由細胞中之處理產生的FAP之任何形式。該術語亦涵蓋FAP之天然存在之變異體,例如剪接變異體或對偶基因變異體。在一個實施例中,本發明之抗原結合分子能夠特異性結合於人類、小鼠及/或獼猴FAP。人類FAP之胺基酸序列展示於UniProt (www.uniprot.org)寄存編號Q12884 (版本149,SEQ ID NO:84)或NCBI (www.ncbi.nlm.nih.gov/)RefSeq NP_004451.2中。人類FAP之胞外域(ECD)自胺基酸位置26延伸至胺基酸位置760。His標記之人類FAP ECD之胺基酸及核苷酸序列分別展示於SEQ ID NO:85及86中。小鼠FAP之胺基酸序列展示於UniProt寄存編號P97321 (版本126,SEQ ID NO:87)或NCBI RefSeq NP_032012.1中。小鼠FAP之胞外域(ECD)自胺基酸位置26延伸至胺基酸位置761。SEQ ID NO:88及89分別展示His標記之小鼠FAP ECD之胺基酸及核苷酸序列。SEQ ID NO:90及91分別展示His標記之獼猴FAP ECD之胺基酸及核苷酸序列。本發明之抗FAP結合分子較佳結合於FAP之胞外域。
除非另有指示,否則術語「
癌胚抗原 ( CEA )」,亦稱為癌胚抗原相關細胞黏附分子5 (CEACAM5),係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生CEA。人類CEA之胺基酸序列展示於UniProt寄存編號P06731 (版本151,SEQ ID NO:92)中。除非另有指示,否則術語「
黑素瘤相關軟骨素硫酸鹽蛋白聚糖 ( MCSP )」,亦稱為軟骨素硫酸鹽蛋白聚糖4 (CSPG4),係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生MCSP。人類MCSP之胺基酸序列展示於UniProt寄存編號Q6UVK1 (版本103,SEQ ID NO:93)中。除非另有指示,否則術語「
表皮生長因子受體 ( EGFR )」,亦稱為原癌基因c-ErbB-1或受體酪胺酸-蛋白質激酶erbB-1,係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生EGFR。人類EGFR之胺基酸序列展示於UniProt寄存編號P00533 (版本211,SEQ ID NO:94)中。除非另有指示,否則術語「
CD19」係指B-淋巴細胞抗原CD19,亦稱為B-淋巴細胞表面抗原B4或T細胞表面抗原Leu-12且包括來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生CD19。人類CD19之胺基酸序列展示於UniProt寄存編號P15391 (版本160,SEQ ID NO:95)中。「
CD20」係指B淋巴細胞抗原CD20,亦稱為跨膜4-結構域子族A成員1 (MS4A1)、B-淋巴細胞表面抗原B1或白細胞表面抗原Leu-16,且包括來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生CD20。人類CD20之胺基酸序列展示於UniProt寄存編號P11836 (版本149,SEQ ID NO:96)中。除非另有指示,否則「
CD33」係指骨髓細胞表面抗原CD33,亦稱為SIGLEC3或gp67,且包括來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生CD33。人類CD33之胺基酸序列展示於UniProt寄存編號P20138 (版本157,SEQ ID NO:97)中。
術語「
可變區」或「可變結構域」係指抗體重鏈或輕鏈中涉及抗原結合分子與抗原之結合的結構域。天然抗體之重鏈及輕鏈(分別為VH及VL)可變域一般具有類似的結構,其中各結構域均包含四個保守性構架區(FR)及三個高變區(HVR)。參看例如Kindt等人, Kuby Immunology, 第6版, W.H. Freeman and Co., 第91頁 (2007)。單一VH或VL域可足以賦予抗原結合特異性。
如本文所用,術語「
高變區」或「HVR」係指抗體可變域中在序列上具有高變性及/或形成結構上定義之環(「高變環」)的各區域。一般而言,原生四鏈抗體包含六個HVR;三個位於VH中(H1、H2、H3),且三個位於VL中(L1、L2、L3)。HVR一般包含來自高變環及/或來自互補決定區(CDR)的胺基酸殘基,後者具有最高的序列可變性及/或與抗原識別相關。例示性高變環出現在胺基酸殘基26-32 (L1)、50-52 (L2)、91-96 (L3)、26-32 (H1)、53-55 (H2)及96-101 (H3)處。(Chothia及Lesk, J. Mol. Biol. 196:901-917 (1987)。) 例示性CDR (CDR-L1、CDR-L2、CDR-L3、CDR-H1、CDR-H2及CDR-H3)出現在L1之胺基酸殘基24-34、L2之胺基酸殘基50-56、L3之胺基酸殘基89-97、H1之胺基酸殘基31-35B、H2之胺基酸殘基50-65及H3之胺基酸殘基95-102處。(Kabat等人, Sequences of Proteins of Immunological Interest, 第5版. Public Health Service, National Institutes of Health, Bethesda, MD (1991)。) 高變區(HVR)亦稱為「互補決定區」(CDR),且此等術語在本文中、在提及形成抗原結合區之可變區之一部分時可互換使用。此具體區域已由Kabat等人, U.S. Dept. of Health and Human Services, "Sequences of Proteins of Immunological Interest" (1983)及Chothia等人, J. Mol. Biol. 196:901-917 (1987)描述,其中定義包括彼此比較時胺基酸殘基之重疊或亞群。然而,涉及抗體或其變異體之CDR之任何定義的應用意欲處於如本文中所定義及所用之術語的範疇內。涵蓋如上文所引用之參考文獻中之每一者所定義之CDR的適當胺基酸殘基闡述於下表A中作為比較。包含特定CDR的確切殘基數目將視CDR序列及大小而變。在抗體之可變區胺基酸序列指定的情況下,熟習此項技術者可以常規方式確定哪個殘基包含特定CDR。
表A.CDR定義
1
1表A中之所有CDR定義之編號皆依據Kabat等人所闡述之編號約定(參看下文)。
2如表A中所用之含有小寫「b」的「AbM」係指如由Oxford Molecular之「AbM」抗體模型化軟體所定義的CDR。
Kabat等人亦定義適用於任何抗體的可變區序列編號系統。一般技術者可將此「Kabat編號」系統明確地分配給任何可變區序列,而不依賴於超過序列本身的任何實驗資料。如本文所用,「Kabat編號」係指由Kabat等人, U.S. Dept. of Health and Human Services, 「Sequence of Proteins of Immunological Interest」 (1983)所闡述之編號系統。除非另外說明,否則提及抗體可變區中之特定胺基酸殘基位置的編號係根據Kabat編號系統。
除VH中之CDR1之外,CDR一般包含形成高變環的胺基酸殘基。CDR亦包含「特異性決定殘基」或「SDR」,其為接觸抗原之殘基。SDR含於簡稱為-CDR或a-CDR之CDR區域內。例示性a-CDR (a-CDR-L1、a-CDR-L2、a-CDR-L3、a-CDR-H1、a-CDR-H2及a-CDR-H3) 出現在L1之胺基酸殘基31-34、L2之胺基酸殘基50-55、L3之胺基酸殘基89-96、H1之胺基酸殘基31-35B、H2之胺基酸殘基50-58及H3之胺基酸殘基95-102。(參看Almagro及Fransson, Front. Biosci. 13:1619-1633 (2008).) 除非另外指示,否則在本文中,根據Kabat等人, 同前文獻對可變域中之HVR殘基及其他殘基(例如FR殘基)進行編號。
「構架」或「FR」係指除高變區(HVR)殘基以外的可變結構域殘基。可變域之FR一般由四個FR域組成:FR1、FR2、FR3及FR4。因此,在VH (或VL)中,HVR及FR序列一般以以下序列呈現:FR1-H1(L1)-FR2-H2(L2)-FR3-H3(L3)-FR4。
術語「
嵌合」抗體係指重鏈及/或輕鏈之一部分衍生自特定來源或物種,而重鏈及/或輕鏈之其餘部分衍生自不同來源或物種之抗體。
抗體之「
類別」係指其重鏈所具有之恆定域或恆定區的類型。存在五種主要類別之抗體:IgA、IgD、IgE、IgG及IgM,且此等類別中之若干者可進一步分成子類(同型),例如IgG
1、IgG
2、IgG
3、IgG
4、IgA
1及IgA
2。對應於不同類別之免疫球蛋白之重鏈恆定域分別稱為α、δ、ε、γ及μ。
「
人類化」抗體係指包含來自非人類HVR之胺基酸殘基及來自人類FR之胺基酸殘基之嵌合抗體。在某些實施例中,人類化抗體將包含至少一個且通常兩個可變域之實質上全部,其中所有或實質上所有HVR (例如CDR)均對應於非人類抗體之HVR,且所有或實質上所有FR均對應於人類抗體之FR。人類化抗體視情況可包含源自人類抗體之抗體恆定區的至少一部分。抗體(例如非人類抗體)之「
人類化形式」係指已經歷人類化之抗體。本發明涵蓋之「人類化抗體」之其他形成為其中恆定區已經額外修飾或自原始抗體發生變化以產生根據本發明之特性,尤其在C1q結合及/或Fc受體(FcR)結合方面。
「
人類」抗體為胺基酸序列對應於由人類或人類細胞產生或來源於利用人類抗體譜系或其他人類抗體編碼序列之非人類來源之抗體之胺基酸序列的抗體。人類抗體之此定義特定排除包含非人類抗原結合殘基之人類化抗體。
術語「Fc結構域」或「
Fc 區」在本文中用於定義抗體重鏈中含有恆定區之至少一部分的C端區域。該術語包括原生序列Fc區及變異型Fc區。IgG Fc區包含IgG CH2及IgG CH3結構域。人類IgG Fc區之「CH2結構域」通常自約位置231處之胺基酸殘基延伸至約位置340處之胺基酸殘基。在一個實施例中,碳水化合物鏈連接至CH2結構域。本文中,CH2結構域可為原生序列CH2結構域或變異型CH2結構域。「CH3結構域」包含Fc區中C端殘基延伸至CH2結構域(亦即自IgG之約位置341之胺基酸殘基至約位置447處之胺基酸殘基)。本文中,CH3區可為原生序列CH3結構域或變異型CH3結構域(例如在其一個鏈中具有引入之「隆凸」(「杵」)且在其另一個鏈中具有相應的引入之「凹穴」(「臼」)的CH3結構域,參看美國專利第5,821,333號,其以引用之方式明確併入本文中)。此類變異型CH3結構域可用於促進如本文中所描述之兩個非一致抗體重鏈之雜二聚化。在一個實施例中,人類IgG重鏈Fc區自Cys226或自Pro230延伸至重鏈之羧基末端。然而,Fc區之C端離胺酸(Lys447)可存在或可不存在。除非本文另外說明,否則Fc區或恆定區胺基酸殘基之編號係依據EU編號系統,亦稱為EU索引,如Kabat等人, Sequences of Proteins of Immunological Interest, 第5版, Public Health Service, National Institutes of Health, Bethesda, MD, 1991中所述。
「
杵 - 臼」技術描述於例如US 5,731,168;US 7,695,936;Ridgway等人, Prot Eng 9, 617-621 (1996)及Carter, J Immunol Meth 248, 7-15 (2001)中。一般而言,該方法涉及在第一多肽之界面處引入隆凸(「杵」)及在第二多肽之界面處引入相應空腔(「臼」),使得隆凸可定位於空腔中以便促進雜二聚體形成且阻礙均二聚體形成。藉由用較大側鏈(例如酪胺酸或色胺酸)置換第一多肽界面中之小胺基酸側鏈來構建隆凸。大小與隆凸相同或相似的補償性空腔藉由用較小胺基酸側鏈(例如丙胺酸或蘇胺酸)置換大胺基酸側鏈而形成於第二多肽之界面中。隆凸及空腔可藉由改變編碼多肽之核酸(例如藉由定點突變誘發或藉由肽合成來改變)來產生。在一個特定實施例中,結修飾包含Fc結構域之兩個子單元中之一者中之胺基酸取代T366W,且孔修飾包含Fc結構域之兩個子單元中之另一者中的胺基酸取代T366S、L368A及Y407V。在另一特定實施例中,包含結修飾之Fc結構域之子單元額外包含胺基酸取代S354C,且包含孔修飾之Fc結構域之子單元額外包含胺基酸取代Y349C。引入此等兩個半胱胺酸殘基使得Fc區之兩個子單元之間形成二硫橋鍵,由此進一步穩定二聚體(Carter, J Immunol Methods 248, 7-15 (2001))。
「與免疫球蛋白之Fc區等效之區域」意欲包括免疫球蛋白之Fc區之天然存在之對偶基因變異體以及具有變化之變異體,該等變化產生取代、添加或缺失,但不實質上降低免疫球蛋白介導效應功能(諸如抗體依賴性細胞毒性)之能力。舉例而言,免疫球蛋白之Fc區之N端或C端可缺失一或多個胺基酸而不實質性損失生物功能。此類變異體可根據此項技術中已知的一般法則選擇以對活性具有最小影響(參看例如Bowie, J. U.等人, Science 247:1306-10 (1990))。
術語「
效應功能」係指可歸因於抗體之Fc區之彼等生物活性,其因抗體同型而異。抗體效應功能之實例包括:C1q結合及補體依賴性細胞毒性(CDC)、Fc受體結合、抗體依賴性細胞介導之細胞毒性(ADCC)、抗體依賴性細胞吞噬(ADCP)、細胞激素分泌、免疫複合物介導抗原呈遞細胞攝入抗原、細胞表面受體(例如B細胞受體)下調及B細胞活化。
可藉由抗體之Fc-區與Fc受體(FcR)相互作用來調節Fc受體結合依賴性效應功能,其為造血細胞上經特殊化之細胞表面受體。Fc受體屬於免疫球蛋白超家族,且已展示經由抗體依賴性細胞毒性(ADCC)其調節包被抗體之病原體之移除(藉由免疫複合體吞噬作用),及紅血球及包被相對應抗體的不同其他細胞靶(例如腫瘤細胞)之裂解兩者 (參看例如,Van de Winkel, J.G.及Anderson, C.L., J. Leukoc. Biol. 49 (1991) 511-524)。FcR由其針對免疫球蛋白同型之特異性定義:針對IgG抗體之Fc受體被稱為FcγR。Ravetch, J.V.及Kinet, J.P., Annu. Rev. Immunol. 9 (1991) 457-492;Capel, P.J.等人, Immunomethods 4 (1994) 25-34;de Haas, M.等人., J. Lab. Clin. Med. 126 (1995) 330-341;及Gessner, J.E.等人, Ann. Hematol. 76 (1998) 231-248描述Fc受體結合。
IgG抗體之Fc區(FcγR)的受體交聯觸發多種效應功能,包括噬菌作用、抗體依賴性細胞毒性及炎性介質釋放,以及免疫複合物清除及抗體產量之調節。在人類中,已表徵三種類別之FcγR,其為:
- FcγRI (CD64)以高親和力結合於單體IgG,且在巨噬細胞、單核細胞、嗜中性白細胞及嗜酸性細胞上表現。在Fc-區IgG內在胺基酸殘基E233-G236、P238、D265、N297、A327及P329 (根據Kabat之EU索引編號)中之一者修飾,減小FcγRI結合。位置233-236處之IgG2殘基(經取代至IgG1及IgG4)減少10³倍FcγR結合,並消除人單核細胞與抗體敏化紅細胞之響應(Armour, K.L.等人, Eur. J. Immunol. 29 (1999) 2613-2624)。
- FcγRII (CD32)以中等至低親和力結合於複合型IgG,且廣泛地表現。此受體可分為兩種次類:FcγRIIA及FcγRIIB。在許多涉及殺滅之細胞(例如,巨噬細胞、單核細胞、嗜中性白細胞)上發現FcγRIIA,且其似乎能夠激活殺滅程序。FcγRIIB似乎在抑制程序中起一定作用,且在B細胞、巨噬細胞上及肥大細胞及嗜酸性細胞上發現FcγRIIB。在B細胞上,其似乎起到遏制進一步產生免疫球蛋白及同型轉換至(例如)IgE類別之功能。在巨噬細胞上,當調節至FcγRIIA時,FcγRIIB用以抑制吞噬作用。在嗜酸性細胞及肥大細胞上,在IgE結合至其獨立受體的整個過程中,B形態可有助於遏制此等細胞。發現FcγRIIA結合減小 ,例如包含至少在胺基酸殘基E233-G236、P238、D265、N297、A327、P329、D270、Q295、A327、R292及K414中之一者處具有突變之IgG Fc-區的抗體(根據Kabat之EU索引編號)。
- FcγRIII (CD16)以中等至低親和力結合於IgG,且以兩種類型存在。在NK細胞、巨噬細胞、嗜酸性細胞及一些單核細胞及T細胞上發現FcγRIIIA,且FcγRIIIA調節ADCC。FcγRIIIB在嗜中性白細胞上高度表現。發現FcγRIIIA結合減小,例如包含至少在胺基酸殘基E233-G236、P238、D265、N297、A327、P329、D270、Q295、A327、S239、E269、E293、Y296、V303、A327、K338及D376中之一者處具有突變之IgG Fc-區的抗體(根據Kabat之EU索引編號)。
在人IgG1上映射Fc受體結合位點、上述突變位點及用於量測FcγRI及FcγRIIA結合之方法描述於Shields, R.L.等人 J. Biol. Chem. 276 (2001) 6591-6604中。
術語「
ADCC」或「抗體依賴性細胞毒性」為藉由Fc受體結合調節之功能,且係指在效應細胞存在下的情況下藉由如本文所報導之抗體裂解目標細胞。藉由量測該等結合Fcγ受體表現細胞,諸如以重組方式表現FcγRI及/或FcγRIIA或NK細胞(基本上表現FcγRIIIA)之細胞,來研究引起調節ADCC之最初步驟的抗體含量。特定言之,量測NK細胞上與FcγR之結合。
「
活化 Fc 受體」為一種Fc受體,其與抗體之Fc區接合之後,引發信號傳導事件,刺激攜帶受體之細胞執行效應功能。活化Fc受體包括FcγRIIIa (CD16a)、FcγRI (CD64)、FcγRIIa (CD32)及FcαRI (CD89)。特定活化Fc受體為人類FcγRIIIa (參看UniProt寄存編號P08637,版本141)。
「腫瘤壞死因子受體超家族」或「
TNF 受體超家族」目前由27受體組成。其為特徵為經富含半胱胺酸之胞外域(CRD)結合腫瘤壞死因子(TNF)之能力細胞激素受體之群。此等偽重複由受體鏈內高度保守半胱胺酸殘基產生之鏈內二硫化物界定。除神經生長因子(NGF)以外,全部TNF與原型TNF-α同源。大部分TNF受體以其活性形式在漿膜中形成三聚複合物。因此,大部分TNF受體含有跨膜結構域(TMD)。若干此等受體亦含有胞內死亡結構域(DD),其在配位體結合之後募集卡斯蛋白酶相互作用蛋白以起始卡斯蛋白酶活化之外源路徑。缺乏死亡結構域之其他TNF超家族受體結合TNF受體相關因子且活化可導致增殖或分化之胞內信號傳導路徑。此等受體亦可起始細胞凋亡,但其經間接機制進行。除了調整細胞凋亡之外,若干TNF超家族受體涉及於調整免疫細胞功能(諸如B細胞穩定及活化)、自然殺手細胞活化及T細胞共刺激。若干其他受體調整細胞類型特異性反應,諸如毛囊發育及蝕骨細胞發育。TNF受體超家族之成員包括以下:腫瘤壞死因子受體1 (1A) (TNFRSF1A、CD120a)、腫瘤壞死因子受體2 (1B) (TNFRSF1B、CD120b)、淋巴毒素β受體(LTBR、CD18)、OX40 (TNFRSF4、CD134)、CD40 (Bp50)、Fas受體(Apo-1、CD95、FAS)、誘餌受體3 (TR6、M68、TNFRSF6B)、CD27 (S152、Tp55)、CD30 (Ki-1、TNFRSF8)、4-1BB (CD137、TNFRSF9)、DR4 (TRAILR1、Apo-2、CD261、TNFRSF10A)、DR5 (TRAILR2、CD262、TNFRSF10B)、誘餌受體1 (TRAILR3、CD263、TNFRSF10C)、誘餌受體2 (TRAILR4、CD264、TNFRSF10D)、RANK (CD265、TNFRSF11A)、骨保護素(OCIF、TR1、TNFRSF11B)、TWEAK受體(Fn14、CD266、TNFRSF12A)、TACI (CD267、TNFRSF13B)、BAFF受體(CD268、TNFRSF13C)、疱疹病毒侵入介體(HVEM、TR2、CD270、TNFRSF14)、神經生長因子受體(p75NTR、CD271、NGFR)、B細胞成熟抗原(CD269、TNFRSF17)、糖皮質激素誘導之TNFR相關(GITR、AITR、CD357、TNFRSF18)、TROY (TNFRSF19)、DR6 (CD358、TNFRSF21)、DR3 (Apo-3、TRAMP、WS-1、TNFRSF25)以及外異蛋白A2受體(XEDAR、EDA2R)。
腫瘤壞死因子受體(TNFR)家族之若干成員在初始T細胞活化之後用於維持T細胞反應。術語「
共刺激 TNF 受體 家族成員」或「共刺激TNF家族受體」係指能夠共刺激T細胞之增殖及細胞激素產生的TNF受體家族成員之子群。除非另外指示,否則該術語係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)、非人類靈長類動物(例如食蟹獼猴)及嚙齒動物(例如小鼠及大鼠))之任何原生TNF家族受體。在本發明之具體實施例中,共刺激TNF受體家族成員係選自由以下組成之群:OX40 (CD134)、4-1BB (CD137)、CD27、HVEM (CD270)、CD30及GITR,其均可對T細胞具有共刺激作用。更特定言之,共刺激TNF受體家族成員選自由OX40及4-1BB組成之群。
TNF受體家族成員之其他資訊(特定言之,序列)可自公開可用資料庫(諸如Uniprot (www.uniprot.org))獲得。舉例而言,人類共刺激TNF受體具有以下胺基酸序列:人類OX40 (UniProt寄存編號P43489,SEQ ID NO:98)、人類4-1BB (UniProt寄存編號Q07011,SEQ ID NO:99)、人類CD27 (UniProt寄存編號P26842,SEQ ID NO:100)、人類HVEM (UniProt寄存編號Q92956,SEQ ID NO:101)、人類CD30 (UniProt寄存編號P28908,SEQ ID NO:102)以及人類GITR (UniProt寄存編號Q9Y5U5,SEQ ID NO:103)。
除非另外指示,否則如本文所用之術語「
OX40」係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)及嚙齒動物(例如小鼠及大鼠))之任何原生OX40。該術語涵蓋「全長」的未經處理之OX40以及由細胞中之處理產生的OX40之任何形式。該術語亦涵蓋OX40之天然存在之變異體,例如剪接變異體或對偶基因變異體。例示性人類OX40之胺基酸序列顯示於SEQ ID NO: 98 (Uniprot P43489,型式112)且例示性鼠類OX40之胺基酸序列顯示於SEQ ID NO: 104 (Uniprot P47741,型式101)中。
術語「
抗 OX40 抗體」、「抗OX40」、「OX40抗體」及「特異性結合於OX40之抗體」係指能夠以足夠親和力結合OX40以使得抗體在靶向OX40時適用作診斷劑及/或治療劑之抗體。在一個實施例中,抗OX40抗體與不相關非OX40蛋白質之結合程度小於例如放射免疫分析(RIA)或流動式細胞量測術(FACS)量測之抗體與OX40之結合的約10%。在某些實施例中,結合於OX40之抗體的解離常數(K
D) ≤ 1 μM、≤ 100 nM、≤ 10 nM、≤ 1 nM、≤ 0.1 nM、≤ 0.01 nM或≤ 0.001 nM (例如10
- 6M或更低,例如10
- 68M至10
- 13M,例如10
- 8M至10
- 10M)。
除非另外指示,否則如本文所用之術語「
4 - 1BB」係指來自任何脊椎動物來源(包括哺乳動物,諸如靈長類動物(例如人類)及嚙齒動物(例如小鼠及大鼠))之任何原生4-1BB。該術語涵蓋「全長」未經處理之4-1BB以及由細胞中之處理產生的4-1BB之任何形式。該術語亦涵蓋4-1BB之天然存在之變異體,例如剪接變異體或對偶基因變異體。例示性人類4-1BB之胺基酸序列顯示於SEQ ID NO: 99 (Uniprot寄存編號Q07011)中,例示性鼠類4-1BB之胺基酸序列顯示於SEQ ID NO: 105 (Uniprot寄存編號P20334)中且例示性獼猴4-1BB (來自恆河猴)之胺基酸序列顯示於SEQ ID NO:106 (Uniprot寄存編號F6W5G6)中。
術語「
抗 4 - 1BB 抗體」、「抗4-1BB」、「4-1BB抗體」及「特異性結合於4-1BB之抗體」係指能夠以足夠親和力結合4-1BB以使得抗體在靶向4-1BB時適用作診斷劑及/或治療劑之抗體。在一個實施例中,抗4-1BB抗體與不相關非4-1BB蛋白質的結合程度低於例如放射免疫分析(RIA)或流動式細胞量測術(FACS)所量測之抗體與4-1BB之結合的約10%。在某些實施例中,結合於4-1BB之抗體的解離常數(K
D) ≤ 1 μM、≤ 100 nM、≤ 10 nM、≤ 1 nM、≤ 0.1 nM、≤ 0.01 nM或≤ 0.001 nM (例如10
- 6M或更低,例如10
- 68M至10
- 13M,例如10
- 8M至10
- 10M)。
術語「
肽連接子」係指包含一或多個胺基酸(通常約2至20個胺基酸)的肽。肽連接子在此項技術中已知或描述於本文中。適合非免疫原性連接肽為例如(G
4S)
n、(SG
4)
n或G
4(SG
4)
n肽連接子,其中「n」一般為介於1與10之間,通常介於2與4之間,具體而言為2之數字,即肽選自由以下組成之群:GGGGS (SEQ ID NO: 107) GGGGSGGGGS (SEQ ID NO:108)、SGGGGSGGGG (SEQ ID NO:109)及GGGGSGGGGSGGGG (SEQ ID NO:110),但亦包括序列GSPGSSSSGS (SEQ ID NO:111)、(G4S)
3(SEQ ID NO:112)、(G4S)
4(SEQ ID NO:113)、GSGSGSGS (SEQ ID NO:114)、GSGSGNGS (SEQ ID NO:115)、GGSGSGSG (SEQ ID NO:116)、GGSGSG (SEQ ID NO:117)、GGSG (SEQ ID NO:118)、GGSGNGSG (SEQ ID NO:119)、GGNGSGSG (SEQ ID NO:120)以及GGNGSG (SEQ ID NO:121)。尤其受關注之肽連接子為(G4S) (SEQ ID NO:107)、(G
4S)
2或GGGGSGGGGS (SEQ ID NO:108)及GSPGSSSSGS (SEQ ID NO:111)。
如本申請案內所用,術語「胺基酸」表示天然存在之羧基α-胺基酸之群,其包含丙胺酸(三字母代碼:ala,一字母代碼:A)、精胺酸(arg,R)、天冬醯胺(asn,N)、天冬胺酸(asp,D)、半胱胺酸(cys,C)、麩醯胺酸(gln,Q)、麩胺酸(glu,E)、甘胺酸(gly,G)、組胺酸(his,H)、異白胺酸(ile,I)、白胺酸(leu,L)、離胺酸(lys,K)、甲硫胺酸(met,M)、苯丙胺酸(phe,F)、脯胺酸(pro,P)、絲胺酸(ser,S)、蘇胺酸(thr,T)、色胺酸(trp,W)、酪胺酸(tyr,Y)及纈胺酸(val,V)。
「
稠合」或「
連接」意謂組分(例如抗體及Fab片段之重鏈)經肽鍵直接或經一或多個肽連接子連接。
相對於參考多肽序列之「
胺基酸序列一致性百分比 (%)」定義為在比對參考多肽序列與候選序列且必要時引入間隙以達成最大序列一致性百分比之後,且在不將保守性取代視為序列一致性之一部分之情況下,候選序列中與參考多肽序列中之胺基酸殘基一致的胺基酸殘基之百分比。出於測定胺基酸序列一致性百分比目地之比對可以使用此項技術範圍內的多種方法例如使用公開可獲得之電腦軟體(諸如BLAST、BLAST-2、ALIGN.SAWI或Megalign (DNASTAR)軟件)實現。熟習此項技術者可測定用於比對序列之適當參數,包括在所比較序列之全長內達成最大比對所需的任何算法。然而,出於本文之目的,使用序列比較電腦程式ALIGN-2產生胺基酸序列一致性百分比值。ALIGN-2序列比較電腦程式由Genentech, Inc.創作,且原始程式碼已隨使用者文件一起提交於美國版權局(U.S.Copyright Office), Washington D.C., 20559,其在美國版權局以美國版權登記號TXU510087登記。ALIGN-2程序可公開獲自Genentech, Inc., South San Francisco, California,或可自原始碼編寫。ALIGN-2程序經編寫可用於UNIX操作系統,包括數位UNIX V4.0D。所有序列比較參數由ALIGN-2程序設定且不變化。在採用ALIGN-2進行胺基酸序列比較之情形下,給定胺基酸序列A與給定胺基酸序列B 之胺基酸序列一致性% (或者,其可表述為與給定胺基酸序列B具有或包含一定胺基酸序列一致性%的給定胺基酸序列A)如下計算:
100乘以分率X/Y
其中X為利用序列比對程式ALIGN-2對A與B進行程式比對時評為一致匹配的胺基酸殘基之數目,且其中Y為B中胺基酸殘基的總數。應瞭解,當胺基酸序列A之長度不等於胺基酸序列B之長度時,A與B之胺基酸序列一致性%將不等於B與A之胺基酸序列一致性%。除非另外特定說明,否則本文中使用之所有胺基酸序列一致性%值均使用ALIGN-2電腦程式如剛剛前一段落中所述獲得。
在某些實施例中,涵蓋本文中提供之含有TNF配位三聚體之抗原結合分子的
胺基酸序列變異體。舉例而言,可能需要改良含有TNF配位三聚體之抗原結合分子之結合親和力及/或其他生物特性。含有TNF配位三聚體之抗原結合分子之胺基酸序列變異體可藉由將適合的修飾引入編碼核苷酸序列之分子或藉由肽合成來製備。此類修飾包括例如在抗體之胺基酸序列內的殘基之缺失及/或插入及/或取代。可製造缺失、插入及取代之任一組合以獲得最終構築體,其限制條件為最終構築體具有所需特徵,例如抗原結合。用於取代性突變誘發之相關位點包括HVR及構架(FR)。保守性取代以標題「較佳取代」提供於表B中且在下文中參考胺基酸側鏈分類(1)至(6)進一步描述。可將胺基酸取代引入相關分子且針對所需活性篩選產物,例如保持/改良之抗原結合、降低之免疫原性或改良之ADCC或CDC。
表B
胺基酸可根據共有側鏈特性進行分組:
(1) 疏水性:正白胺酸、Met、Ala、Val、Leu、Ile;
(2) 中性親水性:Cys、Ser、Thr、Asn、Gln;
(3) 酸性:Asp、Glu;
(4) 鹼性:His、Lys、Arg;
(5) 影響鏈取向之殘基:Gly、Pro;
(6) 芳族:Trp、Tyr、Phe。
非保守取代將必然伴有將此等類別中之一者之成員換成另一個類別。
術語「
胺基酸序列變異體」包括實質性變異體,其中在親本抗原結合分子(例如人類化或人類抗體)之一或多個高變區殘基中存在胺基酸取代。通常,所選擇之用於進一步研究之所得變異體與親本抗原結合分子相比將具有某些生物特性之修飾(例如改良)(例如提高之親和力、降低之免疫原性)及/或將實質上保持親本抗原結合分子之某些生物特性。一種例示性取代型變異體為親和力成熟抗體,其可例如使用基於噬菌體呈現之親和力成熟技術(諸如本文所描述之技術)便利地產生。簡言之,一或多個HVR殘基突變且變異型抗原結合分子在噬菌體上呈現且針對特定生物活性(例如結合親和力)進行篩選。在某些實施例中,一或多個HVR內可存在取代、插入或缺失,只要此類變化不實質上降低抗原結合分子結合抗原之能力即可。舉例而言,可在HVR中進行不實質上降低結合親和力之保守改變(例如如本文所提供之保守取代)。可靶向突變誘發之用於鑑別抗體之殘基或區域之適用方法稱為「丙胺酸掃描突變誘發」,如由Cunningham及Wells (1989) Science, 244:1081-1085所描述。在此方法中,鑑別殘基或目標殘基(例如帶電殘基,諸如Arg、Asp、His、Lys及Glu)之群且由中性或帶負電胺基酸(例如丙胺酸或聚丙胺酸)置換以測定抗體與抗原之相互相用是否受到影響。可在對初始取代顯示功能敏感性之胺基酸位置處引入其他取代。或者或另外,抗原-抗原結合分子之晶體結構複合以鑑別抗體與抗原之間的接觸點。此類接觸殘基及鄰近殘基可作為取代候選物之標靶或排除在取代候選物之外。可篩選變異體以判定其是否含有所需特性。
胺基酸序列插入包括在一個殘基至含有一百個或一百個以上殘基之多肽長度範圍內的胺基及/或羧基端融合物,以及具有單個或多個胺基酸殘基之序列內插入。末端插入的實例包括具有N端甲硫胺醯基殘基的本發明之雙特異性抗原結合分子。分子之其他插入型變異體包括N或C端與多肽之融合,此延長雙特異性抗原結合分子之血清半衰期。
在某些實施例中,本文提供之雙特異性抗原結合分子經改變以提高或降低抗體經糖基化之程度。可藉由改變胺基酸序列使得創造或移除之一或多個糖基化位點便利地獲得分子之糖基化變異體。當含有TNF配位三聚體之抗原結合分子包含Fc區時,可改變與其連接之碳水化合物。由哺乳動物細胞產生之天然抗體通常包含分支鏈雙觸角寡醣,其一般藉由N鍵連接至Fc區之CH2域的Asn297。參看例如Wright等人,
TIBTECH15:26-32 (1997). 寡醣可包括各種碳水化合物,例如甘露糖、N-乙醯基葡糖胺(GlcNAc)、半乳糖及唾液酸,以及連接至雙觸寡醣結構之「主幹」中之GlcNAc的海藻糖。在一些實施例中,可進行含有TNF家族配位三聚體之抗原結合分子中寡糖之修飾以產生具有某些改良之特性的變異體。在一個態樣中,提供本發明之雙特異性抗原結合分子或抗體的變異體,其具有碳水化合物結構且不具有連接(直接或間接)至Fc區的海藻糖。此類海藻糖基化變異體可具有改良之ADCC功能,參看例如美國專利公開案第US 2003/0157108號(Presta, L.)或US 2004/0093621 (Kyowa Hakko Kogyo Co., Ltd)。在另一態樣中,本發明之雙特異性抗原結合分子或抗體的變異體具有對分寡糖,例如其中連接至Fc區的二觸角寡醣經GlcNAc對分。此類變異體可具有降低之海藻糖基化及/或改良之ADCC功能,參看例如WO 2003/011878 (Jean-Mairet等人);美國專利第6,602,684號(Umana等人);及US 2005/0123546 (Umana等人)。亦提供寡糖中之至少一個半乳糖殘基與Fc區連接之抗體變異體。此類抗體變異體可具有改良之CDC功能且描述於例如WO 1997/30087 (Patel等人);WO 1998/58964 (Raju, S.);及WO 1999/22764 (Raju, S.)中。
在某些態樣中,可能需要形成本發明之雙特異性抗原結合分子的半胱胺酸工程改造之變異體,例如「thioMAb」,其中分子之一或多個殘基經半胱胺酸殘基取代。在特定態樣中,經取代之殘基存在於分子之可達位點處。藉由用半胱胺酸取代此等殘基,藉此將反應性硫醇基安置於抗體之可達位點且可用於使抗體與其他部分(諸如藥物部分或連接子-藥物部分)結合以產生免疫結合物。在某些態樣中,以下殘基中之任一者或多者可經半胱胺酸取代:輕鏈之V205 (Kabat編號);重鏈之A118 (EU編號);及重鏈Fc區之S400 (EU編號)。半胱胺酸工程改造之抗原結合分子可例如美國專利第7,521,541號中所述產生。
術語「聚核苷酸」係指分離之核酸分子或構築體,例如信使RNA (mRNA)、病毒源RNA或質體DNA (pDNA)。聚核苷酸可包含習知磷酸二酯鍵或非習知鍵(例如醯胺鍵,諸如肽核酸(PNA)中所發現)。術語「核酸分子」係指聚核苷酸中存在的任何一或多個核酸區段,例如DNA或RNA片段。
「
經分離」核酸分子或聚核苷酸意指已自原生環境中轉移的核酸分子、DNA或RNA。舉例而言,出於本發明之目的,編碼載體中所含之多肽的重組性聚核苷酸視為經分離。分離之聚核苷酸之其他實例包括異質宿主細胞中所維持的重組聚核苷酸或溶液中經純化(部分或實質上)之聚核苷酸。經分離聚核苷酸包括通常含有聚核苷酸分子之細胞中所含的聚核苷酸分子,但聚核苷酸分子存在於染色體外或不同於其天然染色體位置之染色體位置。經分離RNA分子包括本發明的活體內或活體外RNA轉錄物,以及正股及負股形式,及雙股形式。本發明之經分離之聚核苷酸或核酸進一步包括以合成方式產生的此類分子。另外,聚核苷酸或核酸可為或可包括調節元件,諸如啟動子、核糖體結合位點或轉錄終止子。
一種核酸或聚核苷酸的核苷酸序列與本發明之參考核苷酸序列至少例如95%「一致」意指該聚核苷酸之核苷酸序列與參考序列一致,但該聚核苷酸序列相對於參考核苷酸序列可每100個核苷酸中包括至多五個點突變。換言之,為了獲得核苷酸序列與參考核苷酸序列至少95%一致的聚核苷酸,參考序列中至多5%的核苷酸可缺失或經另一核苷酸取代,或參考序列中可插入佔參考序列核苷酸總數至多5%的多個核苷酸。參考序列之此等改變可發生於參考核苷酸序列之5'或3'端位置或彼等末端位置之間的任何位置,此等位置個別地穿插於參考序列殘基中或參考序列內的一或多個鄰近基團中。實際來看,任何特定聚核苷酸序列是否與本發明之核苷酸序列至少80%、85%、90%、95%、96%、97%、98%或99%一致可習知地使用已知電腦程式確定,諸如上文關於多肽所論述的電腦程式(例如ALIGN-2)。
術語「
表現卡匣」係指重組或合成方式產生的聚核苷酸,其具有允許靶細胞中之特定核酸發生轉錄的一系列特定核酸元件。重組表現卡匣可併入質體、染色體、粒線體DNA、質體DNA、病毒或核酸片段中。通常,表現載體之重組表現卡匣部分包括待轉錄的核酸序列及啟動子,以及其他序列。在某些實施例中本發明之表現卡匣包含編碼本發明之雙特異性抗原結合分子或其片段的聚核苷酸序列。
術語「
載體」或「表現載體」與「表現構築體」同義且係指用於引入特定基因且引導該基因表現的DNA分子,該DNA分子與該基因在目標細胞中可操作地連接。該術語包括呈自我複製核酸結構之載體以及併入已引入其之宿主細胞之基因組中的載體。本發明之表現載體包含表現卡匣。表現載體允許大量的穩定mRNA發生轉錄。一旦表現載體進入靶細胞內,則藉由細胞轉錄及/或轉譯機構產生由該基因編碼的核糖核酸分子或蛋白質。在一個實施例中,本發明之表現載體包含表現卡匣,該表現卡匣包含編碼本發明之雙特異性抗原結合分子或其片段的聚核苷酸序列。
術語「
宿主細胞」、「宿主細胞株」及「宿主細胞培養物」可互換使用且係指已引入外源核酸之細胞,包括此類細胞之後代。宿主細胞包括「轉化體」及「轉化細胞」,其包括初級轉化細胞及自其衍生之後代(不考慮繼代次數)。後代之核酸含量與親代細胞可能不完全相同,而是可能含有突變。本文包括針對原始轉型細胞篩選或選擇的具有相同功能或生物活性之突變型後代。宿主細胞為可用於產生本發明之雙特異性抗原結合分子的任何類型之細胞系統。宿主細胞包括培養細胞,例如哺乳動物培養細胞,諸如CHO細胞、BHK細胞、NS0細胞、SP2/0細胞、YO骨髓瘤細胞、P3X63小鼠骨髓瘤細胞、PER細胞、PER.C6細胞或融合瘤細胞、酵母細胞、昆蟲細胞及植物細胞(僅舉數例),而且包括轉殖基因動物、轉殖基因植物或培養植物或動物組織內所含的細胞。
藥劑之「
有效量」係指在所投與之細胞或組織中產生生理變化所需的量。
藥劑(例如醫藥組合物)之「
治療有效量」係指在必需的劑量及時間段情況下,有效達成所要治療或預防結果的量。舉例而言,治療有效量之藥劑消除、減少、延遲、最小化或預防疾病副作用。
「
個體 (individual)」或「個體(subject)」為哺乳動物。哺乳動物包括(但不限於)家養動物(例如牛、綿羊、貓、狗及馬)、靈長類動物(例如人類及非人類靈長類動物,諸如猴)、家兔及嚙齒動物(例如小鼠及大鼠)。特定言之,個體為人類。
術語「
醫藥組合物」係指所呈形式允許其中所含活性成分之生物活性有效發揮的製劑,且其不含對調配物將投與之個體具有不可接受毒性之其他組分。
「
醫藥學上可接受之賦形劑」係指醫藥組合物中之除活性成分以外的對個體無毒的成分。醫藥學上可接受之賦形劑包含(但不限於)緩衝液、穩定劑或防腐劑。
術語「
藥品說明書」用於指通常包括於治療性產品之商業包裝中之說明書,其含有關於與使用此類治療性產品有關之適應症、用法、劑量、投與、組合療法、禁忌症及/或警告的資訊。
如本文所用,「
治療 (treatment)」(及其語法變化形式,諸如「治療(treat)」或「治療(treating)」)係指臨床介入以試圖改變所治療個體之自然病程,且可以為實現預防或在臨床病理學病程中進行。所需治療作用包括(但不限於)預防疾病發生或復發、緩解症狀、減輕疾病之任何直接或間接病理性後果、預防轉移、降低疾病進展速率、改善或緩和疾病病況及緩解或改善預後。在一些實施例中,本發明之分子用於延遲疾病發展或減慢疾病之進程。
如本文所用之術語「
癌症」係指增殖性疾病,諸如淋巴瘤、淋巴球性白血病、肺癌、非小細胞肺(NSCL)癌、支氣管肺泡細胞肺癌、骨癌、胰腺癌、皮膚癌、頭癌或頸癌、皮膚或眼內黑素瘤、子宮癌、卵巢癌、直腸癌、肛門區癌、胃癌(stomach cancer)、胃癌(gastric cancer)、結腸癌、乳癌、子宮癌、輸卵管癌、子宮內膜癌、子宮頸癌、陰道癌、外陰癌、霍奇金氏病(Hodgkin's Disease)、食道癌、小腸癌、內分泌系統癌、甲狀腺癌、副甲狀腺癌、腎上腺癌、軟組織肉瘤、尿道癌、陰莖癌、前列腺癌、膀胱癌、腎癌或輸尿管癌、腎細胞癌、腎盂癌、間皮瘤、肝細胞癌、膽道癌、中樞神經系統(CNS)贅瘤、脊軸腫瘤、腦幹神經膠質瘤、多形性膠質母細胞瘤、星形細胞瘤、神經鞘瘤、室管膜瘤、神經管胚細胞瘤、腦膜瘤、鱗狀細胞癌、垂體腺瘤及尤文氏肉瘤(Ewing's sarcoma),包括任何上述癌症之難治癒型式,或上述癌症中之一或多者的組合。
本發明之雙特異性抗體本發明提供具有尤其有利特性之新穎生物特異性抗原結合分子,諸如生產能力、穩定性、結合親和力、生物活性、靶向效率及降低之毒性。
例示性雙特異性抗原結合分子在一個態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 至少一個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 至少一個能夠特異性結合於目標細胞抗原之部分體,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個特定態樣中,此等雙特異性抗原結合分子特徵在於促效結合於共刺激TNF受體家族成員。特定言之,共刺激TNF受體家族成員選自由OX40及4-1BB組成之群。
結合於 OX40 之雙特異性抗原結合分子在一個態樣中,共刺激TNF受體家族成員為OX40。特定言之,本發明提供雙特異性抗原結合分子,其中能夠特異性結合於共刺激TNF受體家族成員之部分體結合於包含胺基酸序列SEQ ID NO:1之胺基酸序列的多肽。
在一個態樣中,提供雙特異性抗原結合分子,其包含至少一個能夠特異性結合於OX40之部分體,其中該部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:2及SEQ ID NO:3組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:4及SEQ ID NO:5組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO: 9、SEQ ID NO:10、SEQ ID NO:11及SEQ ID NO:12組成之群的胺基酸序列,
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:13、SEQ ID NO:14及SEQ ID NO:15組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:16、SEQ ID NO:17及SEQ ID NO:18組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23及SEQ ID NO:24組成之群的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其包含至少一個能夠特異性結合於OX40之部分體,其中該部分體包含
(a) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:6之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:19之CDR-H3,
(b) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:7之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:20之CDR-H3,
(c) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:8之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:21之CDR-H3,
(d) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:9之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:22之CDR-H3,
(e) VH結構域,其包含:包含胺基酸序列SEQ ID NO:3之CDR-H1、包含胺基酸序列SEQ ID NO:5之CDR-H2、包含胺基酸序列SEQ ID NO:10之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:14之CDR-L1、包含胺基酸序列SEQ ID NO:17之CDR-H2及包含胺基酸序列SEQ ID NO:23之CDR-H3,
(f) VH結構域,其包含:包含胺基酸序列SEQ ID NO:3之CDR-H1、包含胺基酸序列SEQ ID NO:5之CDR-H2、包含胺基酸序列SEQ ID NO:11之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:14之CDR-L1、包含胺基酸序列SEQ ID NO:17之CDR-H2及包含胺基酸序列SEQ ID NO:23之CDR-H3,或
(g) VH結構域,其包含:包含胺基酸序列SEQ ID NO:3之CDR-H1、包含胺基酸序列SEQ ID NO:5之CDR-H2、包含胺基酸序列SEQ ID NO:12之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:15之CDR-L1、包含胺基酸序列SEQ ID NO:18之CDR-H2及包含胺基酸序列SEQ ID NO:24之CDR-H3。
在一個態樣中,本發明提供一種雙特異性抗原結合分子,其包含至少一個能夠特異性結合於OX40之部分體,其中該部分體包含VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:7之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:20之CDR-H3。
在另一態樣中,本發明提供一種雙特異性抗原結合分子,其中能夠特異性結合於OX40之部分體包含重鏈可變區VH,其包含與選自由SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO: 29、SEQ ID NO: 31、SEQ ID NO: 33、SEQ ID NO:35及SEQ ID NO: 37組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區VL,其包含與胺基酸序列SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO: 30、SEQ ID NO: 32、SEQ ID NO: 34、SEQ ID NO:36及SEQ ID NO: 38至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其中能夠特異性結合於OX40之部分體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
在一個特定態樣中,提供雙特異性抗原結合分子,其中能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL。
結合於 4 - 1BB 之雙特異性抗原結合分子在另一態樣中,共刺激TNF受體家族成員為4-1BB。詳言之,本發明提供雙特異性抗原結合分子,其中能夠特異性結合於共刺激TNF受體家族成員之部分體結合於包含胺基酸序列SEQ ID NO:39之多肽。
在一個態樣中,提供雙特異性抗原結合分子,其包含至少一個能夠特異性結合於4-1BB之部分體,其中該部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:40及SEQ ID NO:41組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:42及SEQ ID NO:43組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47及SEQ ID NO:48組成之群的胺基酸序列
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:49及SEQ ID NO:50組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:51及SEQ ID NO:52組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:53、SEQ ID NO:54、SEQ ID NO:55、SEQ ID NO:56及SEQ ID NO:57組成之群的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其包含至少一個能夠特異性結合於4-1BB之部分體,其中該部分體包含
(a) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:44之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:53之CDR-H3,
(b) VH結構域,其包含:包含胺基酸序列SEQ ID NO:41之CDR-H1、包含胺基酸序列SEQ ID NO:43之CDR-H2、包含胺基酸序列SEQ ID NO:45之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:50之CDR-L1、包含胺基酸序列SEQ ID NO:52之CDR-H2及包含胺基酸序列SEQ ID NO:54之CDR-H3,
(c) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:46之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:55之CDR-H3,
(d) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:47之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:56之CDR-H3,或
(e) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:48之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:57之CDR-H3。
在另一態樣中,本發明提供一種雙特異性抗原結合分子,其中能夠特異性結合於4-1BB之部分體包含重鏈可變區VH,其包含與選自由SEQ ID NO:58、SEQ ID NO:60、SEQ ID NO:62、SEQ ID NO:64及SEQ ID NO:66組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與選自由SEQ ID NO:59、SEQ ID NO:61、SEQ ID NO:63、SEQ ID NO:65及SEQ ID NO:67組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其中能夠特異性結合於4-1BB之部分體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
本發明之雙特異性抗原結合分子特徵進一步為包含至少一個能夠特異性結合於目標細胞抗原之部分體。雙特異性抗原結合分子因此具有優於能夠特異性結合於共刺激TNF受體家族成員之習知抗體的優勢,優勢在於其在目標細胞(通常癌細胞)處選擇性誘發共刺激T細胞反應。在一個態樣中,目標細胞抗原選自由以下組成之群:纖維母細胞活化蛋白(FAP)、黑素瘤相關硫酸軟骨素蛋白聚糖(MCSP)、表皮生長因子受體(EGFR)、癌胚抗原(CEA)、CD19、CD20及CD33。
目標細胞抗原為 FAP 之雙特異性抗原結合分子在一個特定態樣中,目標細胞抗原為纖維母細胞活化蛋白(FAP)。FAP結合部分已描述於WO 2012/02006中,其以全文引用之方式納入。尤其受關注之FAP結合部分描述於下文中。
在一個態樣中,本發明提供雙特異性抗原結合分子,其中能夠特異性結合於FAP之部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:68及SEQ ID NO:69組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:70及SEQ ID NO:71組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:72及SEQ ID NO:73組成之群的胺基酸序列,
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:74及SEQ ID NO:75組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:76及SEQ ID NO:77組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:78及SEQ ID NO:79組成之群的胺基酸序列。
特定言之,提供雙特異性抗原結合分子,其中能夠特異性結合於FAP之部分體包含
(a) VH結構域,其包含:包含胺基酸序列SEQ ID NO:68之CDR-H1、包含胺基酸序列SEQ ID NO:70之CDR-H2、包含胺基酸序列SEQ ID NO:72之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:74之CDR-L1、包含胺基酸序列SEQ ID NO:76之CDR-H2及包含胺基酸序列SEQ ID NO:78之CDR-H3,或
(b) VH結構域,其包含:包含胺基酸序列SEQ ID NO:69之CDR-H1、包含胺基酸序列SEQ ID NO:71之CDR-H2、包含胺基酸序列SEQ ID NO:73之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:75之CDR-L1、包含胺基酸序列SEQ ID NO:77之CDR-H2及包含胺基酸序列SEQ ID NO:79之CDR-H3。
在一個特定態樣中,能夠特異性結合於FAP之部分體包含VH結構域,其包含:包含胺基酸序列SEQ ID NO:69之CDR-H1、包含胺基酸序列SEQ ID NO:71之CDR-H2、包含胺基酸序列SEQ ID NO:73之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:75之CDR-L1、包含胺基酸序列SEQ ID NO:77之CDR-H2及包含胺基酸序列SEQ ID NO:79之CDR-H3。
特定言之,提供雙特異性抗原結合分子,其中能夠特異性結合於FAP之部分體包含
(i) 包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區VL,或
(ii) 包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區VL。
結合於OX40
及 FAP之雙
特異性抗原結合分子在另一態樣中,提供雙特異性抗原結合分子,其中
(i) 能夠特異性結合於OX40之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:25、SEQ ID NO: 27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35或SEQ ID NO:37至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:26、SEQ ID NO: 28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQ ID NO:36或SEQ ID NO:38至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及
(ii) 能夠特異性結合於FAP之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:80或SEQ ID NO:82至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:81或SEQ ID NO:83至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
在一個特定態樣中,提供雙特異性抗原結合分子,其中
(a) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(b) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO: 82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(c) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(d) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(e) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(f) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(g) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(h) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(i) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(j) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(k) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(l) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(m) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,或
(n) 能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO: 82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其中能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,或其中能夠特異性結合於OX40之部分體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO: 82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區。
結合於 4 - 1BB 及 FAP 之雙特異性抗原結合分子在另一態樣中,提供雙特異性抗原結合分子,其中
(i) 能夠特異性結合於4-1BB之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:58、SEQ ID NO: 60、SEQ ID NO:62、SEQ ID NO:64或SEQ ID NO:66至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:59、SEQ ID NO: 61、SEQ ID NO:63、SEQ ID NO:65或SEQ ID NO:67至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及
(ii) 能夠特異性結合於FAP之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:80或SEQ ID NO:82至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:81或SEQ ID NO:83至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
在一個特定態樣中,提供雙特異性抗原結合分子,其中
(a) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(b) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:95之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(c) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(d) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(e) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(f) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(g) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,
(h) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區,
(i) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:80之重鏈可變區VH及包含胺基酸序列SEQ ID NO:81之輕鏈可變區,或
(j) 能夠特異性結合於4-1BB之部分體包括包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區,且能夠特異性結合於FAP之部分體包括包含胺基酸序列SEQ ID NO:82之重鏈可變區VH及包含胺基酸序列SEQ ID NO:83之輕鏈可變區。
雙特異性單價抗原結合分子 ( 1 + 1 型式 )在一個態樣中,本發明係關於雙特異性抗原結合分子,其包含(a)一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成之Fc結構域。
在一個特定態樣中,提供雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,
(b) 能夠特異性結合於目標細胞抗原之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個態樣中,提供雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於OX40之第一Fab片段,
(b) 能夠特異性結合於目標細胞抗原之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在另一態樣中,提供雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於OX40之第一Fab片段,
(b) 能夠特異性結合於FAP之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 包含胺基酸序列SEQ ID NO:303之第一重鏈、包含胺基酸序列SEQ ID NO:182之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(b) 包含胺基酸序列SEQ ID NO:231之第一重鏈、包含胺基酸序列SEQ ID NO:186之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(c) 包含胺基酸序列SEQ ID NO:233之第一重鏈、包含胺基酸序列SEQ ID NO:190之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(d) 包含胺基酸序列SEQ ID NO:235之第一重鏈、包含胺基酸序列SEQ ID NO:194之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(e) 包含胺基酸序列SEQ ID NO:237之第一重鏈、包含胺基酸序列SEQ ID NO:198之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(f) 包含胺基酸序列SEQ ID NO:239之第一重鏈、包含胺基酸序列SEQ ID NO:202之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈。
在一個態樣中,提供雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於4-1BB之第一Fab片段,
(b) 能夠特異性結合於目標細胞抗原之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在另一態樣中,提供雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於4-1BB之第一Fab片段,
(b) 能夠特異性結合於FAP之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 包含胺基酸序列SEQ ID NO:295之第一重鏈、包含胺基酸序列SEQ ID NO:261之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(b) 包含胺基酸序列SEQ ID NO:297之第一重鏈、包含胺基酸序列SEQ ID NO:265之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(c) 包含胺基酸序列SEQ ID NO:299之第一重鏈、包含胺基酸序列SEQ ID NO:269之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(d) 包含胺基酸序列SEQ ID NO:301之第一重鏈、包含胺基酸序列SEQ ID NO:273之第一輕鏈、包含胺基酸序列SEQ ID NO:229之第二重鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈。
雙特異性二價抗原結合分子 ( 2 + 2 型式 )在另一態樣中,本發明係關於一種雙特異性抗原結合分子,其包含
(a) 兩個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 兩個能夠特異性結合於目標細胞抗原之部分體,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
在一個態樣中,雙特異性抗原結合分子對於共刺激TNF受體家族成員及目標細胞抗原皆為二價。
在一個態樣中,本發明之雙特異性抗原結合分子包含(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,以及(b)兩個能夠特異性結合於目標細胞抗原之額外Fab片段,其中該等額外Fab片段各自經肽連接子連接至(a)之重鏈的C端。
在一個特定態樣中,肽連接子為(G4S)
4。
在另一態樣中,能夠特異性結合於目標細胞抗原之兩個額外Fab片段為交換Fab片段,其中可變域VL及VH彼此置換且VL-CH鏈各自經肽連接子連接於(a)之重鏈的C端。
特定言之,本發明係關於雙特異性抗原結合分子,其中兩個能夠特異性結合於共刺激TNF受體家族成員之Fab片段為兩個能夠特異性結合於OX40或4-1BB之Fab片段且兩個能夠特異性結合於目標細胞抗原之額外Fab片段為能夠特異性結合於FAP之交換Fab片段。
在一個態樣中,本發明係關於一種雙特異性抗原結合分子,其包含(a)兩個能夠特異性結合於OX40之部分體,(b)兩個能夠特異性結合於FAP之部分體,及(c)由能夠穩定結合之第一及第二子單元構成之Fc結構域。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 各自包含胺基酸序列SEQ ID NO:216之兩個重鏈、包含胺基酸序列SEQ ID NO:182之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(b) 各自包含胺基酸序列SEQ ID NO:219之兩個重鏈、包含胺基酸序列SEQ ID NO:186之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(c) 各自包含胺基酸序列SEQ ID NO:221之兩個重鏈、包含胺基酸序列SEQ ID NO:190之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(d) 各自包含胺基酸序列SEQ ID NO:223之兩個重鏈、包含胺基酸序列SEQ ID NO:194之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(e) 各自包含胺基酸序列SEQ ID NO:225之兩個重鏈、包含胺基酸序列SEQ ID NO:198之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(f) 各自包含胺基酸序列SEQ ID NO:227之兩個重鏈、包含胺基酸序列SEQ ID NO:202之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈。
在一個態樣中,本發明係關於一種雙特異性抗原結合分子,其包含(a)兩個能夠特異性結合於4-1BB之部分體,(b)兩個能夠特異性結合於FAP之部分體,及(c)由能夠穩定結合之第一及第二子單元構成之Fc結構域。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 各自包含胺基酸序列SEQ ID NO:287之兩個重鏈、包含胺基酸序列SEQ ID NO:261之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(b) 各自包含胺基酸序列SEQ ID NO:289之兩個重鏈、包含胺基酸序列SEQ ID NO:265之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(c) 各自包含胺基酸序列SEQ ID NO:291之兩個重鏈、包含胺基酸序列SEQ ID NO:269之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈,或
(d) 各自包含胺基酸序列SEQ ID NO:293之兩個重鏈、包含胺基酸序列SEQ ID NO:273之第一輕鏈及包含胺基酸序列SEQ ID NO:217之第二輕鏈。
對於結合於共刺激 TNF 受體家族成員為二價且對於結合於目標細胞抗原為單價之雙特異性抗原結合分子 ( 2 + 1 型式 )在另一態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a)兩個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 一個能夠特異性結合於目標細胞抗原之部分體,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
因此,提供一種雙特異性抗原結合分子,其中該雙特異性抗原結合分子對於共刺激TNF受體家族成員為二價且對於目標細胞抗原為單價。
在一個特定態樣中,雙特異性抗原結合分子包含
(a) 包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,及
(b) 能夠特異性結合於目標細胞抗原之VH及VL結構域,其中VH結構域經肽連接子連接至重鏈中之一者的C端且其中VL結構域經肽連接子連接至第二重鏈之C端。
在另一特定態樣中,雙特異性抗原結合分子包含
(a) 包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,及
(b) 能夠特異性結合於目標細胞抗原之VH及VL結構域,其中VH結構域經肽連接子連接於Fc杵重鏈之C端且其中VL結構域經肽連接子連接於Fc臼重鏈之C端。
在另一特定態樣中,雙特異性抗原結合分子包含
(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,及
(b)能夠特異性結合於目標細胞抗原之VH及VL結構域,其中VH結構域經肽連接子連接於Fc臼重鏈之C端,且其中VL結構域經肽連接子連接於Fc杵重鏈之C端。特定言之本發明係關於雙特異性抗原結合分子,其中能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段為兩個能夠特異性結合於OX40或4-1BB之Fab片段及能夠特異性結合於目標細胞抗原之VH及VL結構域,該目標細胞抗原能夠特異性結合於FAP。
在一個態樣中,本發明係關於一種雙特異性抗原結合分子,其包含
(a) 兩個能夠特異性結合於OX40之Fab片段,(b)能夠特異性結合於FAP之VH及VL結構域,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 各自包含胺基酸序列SEQ ID NO:186之兩個輕鏈、包含胺基酸序列SEQ ID NO:306之第一重鏈及包含胺基酸序列SEQ ID NO:307之第二重鏈,或
(b) 各自包含胺基酸序列SEQ ID NO:186之兩個輕鏈、包含胺基酸序列SEQ ID NO:310之第一重鏈及包含胺基酸序列SEQ ID NO:311之第二重鏈。
在一個態樣中,本發明係關於一種雙特異性抗原結合分子,其包含
(a)兩個能夠特異性結合於4-1BB之Fab片段,(b)能夠特異性結合於FAP之VH及VL結構域,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域。
在一個特定態樣中,本發明提供一種雙特異性抗原結合分子,其包含
(a) 各自包含胺基酸序列SEQ ID NO:261之兩個輕鏈、包含胺基酸序列SEQ ID NO:318之第一重鏈及包含胺基酸序列SEQ ID NO:319之第二重鏈,或
(b) 各自包含胺基酸序列SEQ ID NO:265之兩個輕鏈、包含胺基酸序列SEQ ID NO:322之第一重鏈及包含胺基酸序列SEQ ID NO:323之第二重鏈,或
(c) 各自包含胺基酸序列SEQ ID NO:269之兩個輕鏈、包含胺基酸序列SEQ ID NO:326之第一重鏈及包含胺基酸序列SEQ ID NO:327之第二重鏈,或
(d) 各自包含胺基酸序列SEQ ID NO:273之兩個輕鏈、包含胺基酸序列SEQ ID NO:330之第一重鏈及包含胺基酸序列SEQ ID NO:331之第二重鏈,或
(e) 各自包含胺基酸序列SEQ ID NO:261之兩個輕鏈、包含胺基酸序列SEQ ID NO:334之第一重鏈及包含胺基酸序列SEQ ID NO:335之第二重鏈,或
(f) 各自包含胺基酸序列SEQ ID NO:265之兩個輕鏈、包含胺基酸序列SEQ ID NO:338之第一重鏈及包含胺基酸序列SEQ ID NO:339之第二重鏈,或
(g) 各自包含胺基酸序列SEQ ID NO:269之兩個輕鏈、包含胺基酸序列SEQ ID NO:342之第一重鏈及包含胺基酸序列SEQ ID NO:343之第二重鏈,或
(h) 各自包含胺基酸序列SEQ ID NO:273之兩個輕鏈、包含胺基酸序列SEQ ID NO:346之第一重鏈及包含胺基酸序列SEQ ID NO:347之第二重鏈。
降低 Fc 受體結合及 / 或效應功能之 Fc 結構域修飾本發明之雙特異性抗原結合分子進一步包含由能夠穩定結合之第一及第二子單元構成之Fc結構域。
在某些態樣中,可將一或多個胺基酸修飾引入至本文所提供之抗體的Fc區中,從而產生Fc區變異體。Fc區變異體可包含人類Fc區序列(例如人類IgG1、IgG2、IgG3或IgG4 Fc區),其在一或多個胺基酸位置處包含胺基酸修飾(例如取代)。
Fc結構域賦予本發明之雙特異性抗體有利的藥物動力學特性,包括長血清半衰期,其促進目標組織中之良好聚集及有利的組織-血液分佈率。然而,其同時可引起本發明之雙特異性抗體不合需要地靶向表現Fc受體之細胞並非較佳的攜有抗原之細胞。因此,在特定實施例中,本發明之雙特異性抗體與原生IgG Fc結構域(特定言之IgG1 Fc結構域或IgG4 Fc結構域)相比展現降低之對Fc結構域的結合親和力及/或降低之效應功能。更特定言之,Fc結構域為IgG1 Fc結構域。
在一個此類態樣中,Fc結構域(或包含該Fc結構域的本發明之雙特異性抗原結合分子)與原生IgG1 Fc結構域(或包含原生IgG1 Fc結構域的本發明之雙特異性抗原結合分子)相比展現低於50%,較佳低於20%,更佳低於10%且最佳低於5%的與Fc受體之結合親和力,及/或與原生IgG1 Fc結構域(或包含原生IgG1 Fc結構域的本發明之雙特異性抗原結合分子)相比低於50%、較佳低於20%、更佳低於10%且最佳低於5%效應功能。在一個態樣中,Fc結構域(或包含該Fc結構域的本發明之雙特異性抗原結合分子)不會實質上結合於Fc受體及/或誘發效應功能。在一個特定態樣中,Fc受體為Fcγ受體。在一個態樣中,Fc受體為人類Fc受體。在一個態樣中,Fc受體為活化Fc受體。在一個特定態樣中,Fc受體為活化人類Fcγ受體,更特定言之,人類FcγRIIIa、FcγRI或FcγRIIa,最特定言之,人類FcγRIIIa。在一個態樣中,Fc受體為抑制Fc受體。在一特定態樣中,Fc受體為抑制人類Fcγ受體,更具體言之人類FcγRIIB。在一個態樣中,效應功能為CDC、ADCC、ADCP及細胞激素分泌中之一或多者。在一個特定態樣中,效應功能為ADCC。在一個態樣中,Fc結構域對新生兒Fc受體(FcRn)展現的結合親和力與原生IgG1
Fc結構域相比實質上類似。當Fc結構域(或包含該Fc結構域的本發明之雙特異性抗原結合分子)展現大於約70%,特定言之大於約80%,更特定言之大於約90%的原生IgG1 Fc結構域(或包含原生IgG1 Fc結構域的本發明之雙特異性抗原結合分子)與FcRn的結合親和力時,實現實質上類似的與FcRn之結合。
在一個特定態樣中,相較於未經工程改造之Fc結構域,Fc結構域經工程改造從而對Fc受體的結合親和力減小及/或效應功能降低。在一個特定態樣中,本發明之雙特異性抗原結合分子的Fc結構域包含一或多個降低Fc結構域對Fc受體之結合親和力及/或效應功能的胺基酸突變。通常,Fc結構域之兩個子單元中之每一者中存在相同的一或多個胺基酸突變。在一個態樣中,胺基酸突變使Fc結構域對Fc受體的結合親和力減小。在另一態樣中,胺基酸突變使Fc結構域對Fc受體的結合親和力減小至少2倍、至少5倍或至少10倍。在一個態樣中,包含經工程改造之Fc結構域的本發明之雙特異性抗原結合分子與包含未經工程改造之Fc結構域的本發明之雙特異性抗體相比展現小於20%,特定言之小於10%,更特定言之小於5%的對Fc受體之結合親和力。在一個特定態樣中,Fc受體為Fcγ受體。在其他態樣中,Fc受體為人類Fc受體。在一個態樣中,Fc受體為抑制Fc受體。在一特定態樣中,Fc受體為抑制人類Fcγ受體,更具體言之人類FcγRIIB。在一些態樣中,Fc受體為活化Fc受體。在一個特定態樣中,Fc受體為活化人類Fcγ受體,更特定言之,人類FcγRIIIa、FcγRI或FcγRIIa,最特定言之,人類FcγRIIIa。較佳地,與此等受體中之每一者的結合減少。在一些態樣中,相對於補體組分的結合親和力,即對C1q的特異性結合親和力,亦減小。在一個態樣中,對新生兒Fc受體(FcRn)的結合親和力未減小。當Fc結構域(或包含該Fc結構域的本發明之雙特異性抗原結合分子)對FcRn展現的結合親和力大於Fc結構域之未經工程改造形式(或包含該Fc結構域之未經工程改造形式的本發明之雙特異性抗原結合分子)之結合親和力的約70%時,對FcRn達成實質上相似的結合,即保持Fc結構域對該受體的結合親和力。Fc結構域或包含該Fc結構域的本發明之雙特異性抗原結合分子可展現大於約80%以及甚至大於約90%此類親和力。在某些實施例中,本發明之雙特異性抗原結合分子的Fc結構域經工程改造以具有與非工程改造之Fc結構域相比降低之效應功能。降低之效應功能可包括(但不限於)以下中之一或多者:降低之補體依賴性細胞毒性(CDC)、降低之抗體依賴性細胞介導之細胞毒性(ADCC)、降低之抗體依賴性細胞吞噬(ADCP)、降低之細胞激素分泌、降低之免疫複合物介導之抗原呈現細胞之抗原捕捉、降低之與NK細胞之結合、降低之與巨噬細胞之結合、降低之與單核細胞之結合、降低之與多形核細胞之結合、降低之誘導細胞凋亡之直接信號傳導、降低之樹突狀細胞成熟或降低之T細胞激活。
效應功能減小之抗體包括具有Fc區殘基238、265、269、270、297、327及329中之一或多者之取代的彼等抗體(美國專利第6,737,056號)。此類Fc突變異體包括具有胺基酸位置265、269、270、297及327中之兩者或兩者以上之取代的Fc突變異體,包括殘基265及297取代為丙胺酸的所謂「DANA」Fc突變異體(美國專利第7,332,581號)。描述具有提高或降低之與FcR之結合的某些抗體變異體。(例如美國專利第6,737,056號;WO 2004/056312,及Shields, R.L.等人, J. Biol. Chem. 276 (2001) 6591-6604)。
在本發明之一個態樣中,Fc結構域包含在選自E233、L234、L235、N297、P331及P329之位置處的胺基酸取代。在一些態樣中,Fc結構域包含胺基酸取代L234A及L235A (「LALA」)。在一個此類實施例中,Fc結構域為IgG1 Fc結構域,特定言之人類IgG1 Fc結構域。在一個態樣中,Fc結構域包含在位置P329處之胺基酸取代。在一更具體態樣中,胺基酸取代為P329A或P329G,特定言之P329G。在一個實施例中,Fc結構域包含位置P329處之胺基酸取代及選自由以下組成之群的另一胺基酸取代:E233P、L234A、L235A、L235E、N297A、N297D或P331S。在更特定實施例中,Fc結構域包含胺基酸突變L234A、L235A及P329G (「P329G LALA」)。胺基酸取代之「P329G LALA」組合幾乎完全消除人類IgG1 Fc結構域的Fcγ受體結合,如PCT專利申請案第WO 2012/130831 A1號中所述。該文獻亦描述製備此類突變Fc結構域之方法及測定其特性(諸如Fc受體結合或效應功能)的方法。此類抗體為具有突變L234A及L235A或具有突變L234A、L235A及P329G之IgG1 (根據Kabat等人之EU索引、Kabat等人, Sequences of Proteins of Immunological Interest, 第5版. Public Health Service, National Institutes of Health, Bethesda, MD, 1991)。
在一個態樣中,Fc結構域為IgG4
Fc結構域。在一更特定實施例中,Fc結構域為包含位置S228 (Kabat編號)處之胺基酸取代(特定言之,胺基酸取代S228P)的IgG4 Fc結構域。在一個更特定實施例中,Fc結構域為IgG4 Fc結構域,其包含胺基酸取代L235E及S228P及P329G。此胺基酸取代減少活體內IgG4抗體之Fab臂交換(參看Stubenrauch等人, Drug Metabolism and Disposition 38, 84-91 (2010))。
半衰期延長且與負責將母體IgG轉移至胎兒之新生兒Fc受體(FcRn)(Guyer, R.L.等人, J. Immunol. 117 (1976) 587-593;及Kim, J.K.等人, J. Immunol. 24 (1994) 2429-2434)之結合改良之抗體描述於US 2005/0014934中。彼等抗體包含具有一或多個取代之Fc區,該等取代改良Fc區與FcRn之結合。此類Fc變異體包括在以下Fc區殘基中之一或多者處具有取代之彼等變異體:238、256、265、272、286、303、305、307、311、312、317、340、356、360、362、376、378、380、382、413、424或434,例如Fc區殘基434之取代(美國專利第7,371,826號)。Fc區變異體之其他實例亦參看Duncan, A.R.及Winter, G., Nature 322 (1988) 738-740;US 5,648,260;US 5,624,821;及WO 94/29351。
與Fc受體的結合可容易測定,例如藉由ELISA,或藉由表面電漿子共振(SPR)、使用標準儀器,諸如BIAcore儀器(GE Healthcare),且諸如可藉由重組表現來獲得Fc受體。適合的此類結合分析描述於本文中。或者,Fc結構域或包含Fc結構域之細胞活化雙特異性抗原結合分子對Fc受體的結合親和力可使用已知表現特定Fc受體的細胞株(諸如表現FcγIIIa受體的人類NK細胞)評價。Fc結構域或包含Fc結構域之本發明之雙特異性抗體之效應功能可藉由此項技術中已知之方法量測。適用於量測ADCC之分析描述於本文中。用於評定相關分子之ADCC活性的活體外分析之其他實例描述於美國專利第5,500,362號;Hellstrom等人 Proc Natl Acad Sci USA 83, 7059-7063 (1986)及Hellstrom等人, Proc Natl Acad Sci USA 82, 1499-1502 (1985);美國專利第5,821,337號;Bruggemann等人, J Exp Med 166, 1351-1361 (1987)中。或者,可採用非放射性分析方法(參看例如流動式細胞量測術用的ACTI™非放射性細胞毒性分析(CellTechnology, Inc. Mountain View, CA);及CytoTox 96
®非放射性細胞毒性分析(Promega, Madison, WI))。適用於此類分析之效應細胞包括周邊血液單核細胞(PBMC)及天然殺手(NK)細胞。或者或另外,可評定相關分子之活體內ADCC活性,例如在動物模型中,諸如Clynes等人, Proc Natl Acad Sci USA 95, 652-656 (1998)中所揭示之動物模型。
以下部分描述本發明之雙特異性抗原結合分子的較佳態樣,其包含降低Fc受體結合及/或效應功能之Fc結構域修飾。在一個態樣中,本發明係關於雙特異性抗原結合分子(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域,其中Fc結構域包含一或多個降低抗體與Fc受體,特定言之對Fcγ受體之結合親和力的胺基酸取代。在另一態樣中,本發明係關於雙特異性抗原結合分子,其包含(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域,其中Fc結構域包含一或多個降低效應功能之胺基酸取代。在具體態樣中,Fc結構域為具有胺基酸突變L234A、L235A及P329G(根據Kabat EU索引編號)的人類IgG1子類。
促進雜二聚化之 Fc 結構域修飾本發明之雙特異性抗原結合分子包含融合至Fc結構域之兩個子單元的一個或另一個的不同抗原結合位點,因此Fc結構域之兩個子單元可包含兩個不相同多肽鏈。此等多肽之重組共表現及隨後二聚化引起兩種多肽出現若干種可能的組合。為了提高重組製造中本發明之雙特異性抗體的產量及純度,因此將適宜在本發明之雙特異性抗原結合分子的Fc結構域中引入促進所要多肽結合之修飾。
因此,在特定態樣中,本發明係關於雙特異性抗原結合分子,其包含(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域,其中Fc結構域包含促進Fc結構域的第一及第二子單元結合之修飾。人類IgG Fc結構域中之兩個亞單元之間最廣泛蛋白質-蛋白質相互作用的位點存在於Fc結構域之CH3域中。因此,在一個實施例中,該修飾存在於Fc結構域之CH3結構域中。
在一個特定態樣中,該修飾為所謂的「結進孔」修飾,其包含Fc結構域之兩個子單元中之一者中的「杵」修飾及Fc結構域之兩個子單元之另一者中的「臼」修飾。因此,本發明係關於雙特異性抗原結合分子,其包含(a)至少一個能夠特異性結合於共刺激TNF受體家族成員之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域,其中根據杵-臼方法,Fc結構域之第一子單元包含杵且Fc結構域之第二子單元包含臼。在一個特定態樣中,Fc結構域之第一子單元包含胺基酸取代S354C及T366W (EU編號),且Fc結構域之第二子單元包含胺基酸取代Y349C、T366S及Y407V (根據Kabat EU索引編號)。
杵-臼技術描述於例如US 5,731,168;US 7,695,936;Ridgway等人, Prot Eng 9, 617-621 (1996)及Carter, J Immunol Meth 248, 7-15 (2001)中。一般而言,該方法涉及在第一多肽之界面處引入隆凸(「杵」)及在第二多肽之界面處引入相應空腔(「臼」),使得隆凸可定位於空腔中以便促進雜二聚體形成且阻礙均二聚體形成。藉由用較大側鏈(例如酪胺酸或色胺酸)置換第一多肽界面中之小胺基酸側鏈來構建隆凸。大小與隆凸相同或相似的補償性空腔藉由用較小胺基酸側鏈(例如丙胺酸或蘇胺酸)置換大胺基酸側鏈而形成於第二多肽之界面中。
因此,在一個態樣中,在本發明之雙特異性抗原結合分子的Fc結構域的第一子單元的CH3結構域中,胺基酸殘基置換為具有較大側鏈體積之胺基酸殘基,由此在第一子單元之CH3結構域內產生隆凸,其可定位於第二子單元之CH3結構域內的空腔中,且在Fc結構域之第二子單元的CH3結構域中,胺基酸殘基置換為具有較小側鏈體積之胺基酸殘基,由此在第二子單元之CH3結構域內產生空腔,第一子單元之CH3結構域內的隆凸可可定位於該空腔中。隆凸及空腔可藉由改變編碼多肽之核酸(例如藉由定點突變誘發或藉由肽合成來改變)來產生。在一個特定態樣中,在Fc結構域之第一子單元之CH3結構域中,位置366處之蘇胺酸殘基經色胺酸殘基(T366W)置換,且在Fc結構域之第二子單元之CH3結構域中,位置407處之酪胺酸殘基經纈胺酸殘基(Y407V)置換。在一個態樣中,在Fc結構域之第二子單元中,位置366處之蘇胺酸殘基另外經絲胺酸殘基(T366S)置換且位置368處之白胺酸殘基經丙胺酸殘基(L368A)置換。
在另一態樣中,在Fc結構域之第一子單元中,位置354處之絲胺酸殘基另外經半胱胺酸殘基(S354C)置換,且在Fc結構域之第二子單元中,位置349處之酪胺酸殘基另外經半胱胺酸殘基(Y349C)置換。引入此等兩個半胱胺酸殘基使得Fc結構域之兩個子單元之間形成二硫橋鍵,進一步穩定二聚體(Carter (2001), J Immunol Methods 248, 7-15)。在一個特定態樣中,Fc結構域之第一子單元包含胺基酸取代S354C及T366W (EU編號),且Fc結構域之第二子單元包含胺基酸取代Y349C、T366S及Y407V (根據Kabat EU索引編號)。
在一個替代性態樣中,促進Fc結構域之第一及第二子單元之結合的修飾包含調節靜電導引作用之修飾,例如PCT公開案WO 2009/089004中所描述。一般而言,此方法涉及用帶電胺基酸殘基置換兩個Fc結構域子單元之界面處之一或多個胺基酸殘基,使得均二聚體形成變成在靜電方面不利的,但雜二聚在靜電上為有利的。
如本文所報導之雙特異性抗體之重鏈的C端可為以胺基酸殘基PGK結束之全部C端。重鏈之C端可為縮短C端,其中已移除一個或兩個C端胺基酸殘基。在一個較佳態樣中,重鏈之C端為結束PG之縮短C端。在如本文所報導之全部態樣中之一個態樣中,如本文所規定之包含包括C端CH3結構域之重鏈的雙特異性抗體包含C端甘胺酸-離胺酸二肽(G446及K447,根據Kabat EU索引編號)。在如本文所報導之全部態樣中之一個實施例中,如本文所規定之包含包括C端CH3結構域之重鏈的雙特異性抗體包含C端甘胺酸殘基(G446,根據Kabat EU索引編號)。
Fab 結構域中之修飾在一個態樣中,本發明係關於一種雙特異性抗原結合分子,其包含(a)能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,(b)能夠特異性結合於目標細胞抗原之第二Fab片段,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域,其中在Fab片段中之一者中,可變域VH及VL或恆定域CH1及CL經交換。根據互換單抗技術製備雙特異性抗體。
在一個結合臂中具有結構域置換/交換的多特異性抗體(CrossMabVH-VL或CrossMabCH-CL)詳細描述於WO2009/080252及Schaefer, W.等人, PNAS, 108 (2011) 11187-1191中。其明顯地減少由針對第一抗原之輕鏈與針對第二抗原之錯誤重鏈失配(與無此類域交換之方法相比)而造成的副產物。
在一個態樣中,本發明係關於一種雙特異性抗原結合分子,其包含(a)能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,(b)能夠特異性結合於目標細胞抗原之第二Fab片段,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域,其中在Fab片段中之一者中,恆定域CL及CH1彼此交換使得CH1結構域為輕鏈之部分體且CL結構域為重鏈之部分體。更特定言之,在能夠特異性結合於目標細胞抗原之第二Fab片段中,恆定域CL及CH1彼此置換使得CH1結構域為輕鏈之部分體且CL結構域為重鏈之部分體。
在一個特定態樣中,本發明涉及一種雙特異性抗原結合分子,其包含(a)能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,(b)能夠特異性結合於目標細胞抗原之第二Fab片段,其中恆定域CL及CH1彼此置換使得CH1結構域為輕鏈之部分體且CL結構域為重鏈之部分體。此類分子稱為單價雙特異性抗原結合分子。
在另一態樣中,本發明係關於一種雙特異性抗原結合分子,其包含(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,以及(b)兩個能夠特異性結合於目標細胞抗原之額外Fab片段,其中該等額外Fab片段各自經肽連接子連接至(a)之重鏈的C端。在一個特定態樣中,額外Fab片段為可變域VL及VH彼此置換使得VH結構域為輕鏈之部分體且VL結構域為重鏈之部分體的Fab片段。
因此,在一個特定態樣中,本發明包含雙特異性抗原結合分子,其包含(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,及(b)兩個能夠特異性結合於目標細胞抗原之額外Fab片段,其中該兩個能夠特異性結合於目標細胞抗原之額外Fab片段為交換Fab片段,其中可變域VL及VH彼此置換且VL-CH鏈各自經肽連接子連接於(a)之重鏈的C端。
在另一態樣中且進一步改良校正配對,包含以下之雙特異性抗原結合分子:(a)能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,(b)能夠特異性結合於目標細胞抗原之第二Fab片段及(c)由能夠穩定結合之第一及第二子單元構成之Fc結構域,可含有不同帶電荷胺基酸(所謂的「帶電殘基」)取代。將此等修飾引入交叉或非交叉CH1及CL結構域中。在一個特定態樣中,本發明係關於一種雙特異性抗原結合分子,其中在CL結構域中之一者中,位置123 (EU編號)處之胺基酸經精胺酸(R)置換且位置124 (EU編號)處之胺基酸經離胺酸(K)取代,且其中在一個CH1結構域中,位置147 (EU編號)及位置213 (EU編號)處之胺基酸經麩胺酸(E)取代。
更特定言之,本發明係關於包含Fab之雙特異性結合分子,其中在與TNF配位體家族成員相鄰之CL結構域中,位置123 (EU編號)處之胺基酸由精胺酸(R)置換且位置124 (EU編號)處之胺基酸經離胺酸(K)取代,且其中在與TNF配位體家族成員相鄰之CH1結構域中,位置147 (EU編號)及位置213 (EU編號)處之胺基酸經麩胺酸(E)取代。
本發明之例示性抗體在一個態樣中,本發明提供特異性結合於OX40之新抗體及抗體片段。此等抗體具有優於已知OX40抗體之特性,使其尤其適於併入包含能夠特異性結合於目標細胞抗原之另一抗原結合部分的雙特異性抗原結合分子中。
特定言之,提供一種特異性結合於OX40之抗體,其中該抗體包含
(a) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:6之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:19之CDR-H3,
(b) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:7之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:20之CDR-H3,
(c) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:8之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:21之CDR-H3,
(d) VH結構域,其包含:包含胺基酸序列SEQ ID NO:2之CDR-H1、包含胺基酸序列SEQ ID NO:4之CDR-H2、包含胺基酸序列SEQ ID NO:9之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:13之CDR-L1、包含胺基酸序列SEQ ID NO:16之CDR-H2及包含胺基酸序列SEQ ID NO:22之CDR-H3,
(e) VH結構域,其包含:包含胺基酸序列SEQ ID NO:3之CDR-H1、包含胺基酸序列SEQ ID NO:5之CDR-H2、包含胺基酸序列SEQ ID NO:10之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:14之CDR-L1、包含胺基酸序列SEQ ID NO:17之CDR-H2及包含胺基酸序列SEQ ID NO:23之CDR-H3,
(f) VH結構域,其包含:包含胺基酸序列SEQ ID NO:3之CDR-H1、包含胺基酸序列SEQ ID NO:5之CDR-H2、包含胺基酸序列SEQ ID NO:11之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:14之CDR-L1、包含胺基酸序列SEQ ID NO:17之CDR-H2及包含胺基酸序列SEQ ID NO:23之CDR-H3,或
(g) VH結構域,其包含:包含胺基酸序列SEQ ID NO:3之CDR-H1、包含胺基酸序列SEQ ID NO:5之CDR-H2、包含胺基酸序列SEQ ID NO:12之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:15之CDR-L1、包含胺基酸序列SEQ ID NO:18之CDR-H2及包含胺基酸序列SEQ ID NO:24之CDR-H3。
在一個態樣中,提供一種特異性結合於OX40之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
在另一態樣中,提供與特異性結合於OX40之抗體競爭結合之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
在一個態樣中,提供與特異性結合於OX40之抗體競爭結合之抗體,其中該抗體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL。特定言之,提供特異性結合於OX40之抗體,其中該抗體包括包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL。
在另一態樣中,提供特異性結合於OX40且對人類及鼠類OX40具有交叉反應之抗體,其中該抗體包括包含胺基酸序列SEQ ID NO: 31之重鏈可變區VH及包含胺基酸序列SEQ ID NO: 32之輕鏈可變區VL。
在另一態樣中,本發明提供特異性結合於4-1BB之新抗體及抗體片段。此等抗體具有優於已知4-1BB抗體之特性,使得其尤其適於併入包含能夠特異性結合於目標細胞抗原之另一抗原結合部分的雙特異性抗原結合分子中。
特定言之,提供一種特異性結合於4-1BB之抗體,其中該抗體包含
(a) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:44之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:53之CDR-H3,
(b) VH結構域,其包含:包含胺基酸序列SEQ ID NO:41之CDR-H1、包含胺基酸序列SEQ ID NO:43之CDR-H2、包含胺基酸序列SEQ ID NO:45之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:50之CDR-L1、包含胺基酸序列SEQ ID NO:52之CDR-H2及包含胺基酸序列SEQ ID NO:54之CDR-H3,
(c) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:46之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:55之CDR-H3,
(d) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:47之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:56之CDR-H3,或
(e) VH結構域,其包含:包含胺基酸序列SEQ ID NO:40之CDR-H1、包含胺基酸序列SEQ ID NO:42之CDR-H2、包含胺基酸序列SEQ ID NO:48之CDR-H3;及VL結構域,其包含:包含胺基酸序列SEQ ID NO:49之CDR-L1、包含胺基酸序列SEQ ID NO:51之CDR-H2及包含胺基酸序列SEQ ID NO:57之CDR-H3。
在一個態樣中,本發明提供一種特異性結合於4-1BB之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
在另一態樣中,本發明提供與特異性結合於4-1BB之抗體競爭結合之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
聚核苷酸本發明進一步提供編碼如本文所述之雙特異性抗原結合分子的經分離之聚核苷酸或其片段。
編碼本發明之雙特異性抗體的經分離之聚核苷酸可以表現為編碼整個抗原結合分子之單個聚核苷酸或共表現之多個(例如兩個或更多個)聚核苷酸。由共表現之聚核苷酸編碼之多肽可經例如二硫鍵或其他手段結合,以形成功能性抗原結合分子。舉例而言,免疫球蛋白之輕鏈部分可由來自免疫球蛋白之重鏈部分的單獨聚核苷酸編碼。當共表現時,重鏈多肽與輕鏈多肽結合以形成免疫球蛋白。
在一些態樣中,經分離之聚核苷酸編碼如本文所述的根據本發明之雙特異性分子中所包含之多肽。
在一個態樣中,本發明係針對一種編碼雙特異性抗原結合分子之經分離聚核苷酸,其包含(a)至少一個能夠特異性結合於OX40之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域。
在另一態樣中,提供一種編碼雙特異性抗原結合分子之經分離聚核苷酸,其包含(a)至少一個能夠特異性結合於4-1BB之部分體,(b)至少一個能夠特異性結合於目標細胞抗原之部分體,及(c)由能夠穩定結合之第一及第二子單元構成的Fc結構域。
在另一態樣中,本發明係針對一種經分離聚核苷酸,其包含編碼在OX40處特異性結合之抗體或抗體片段的序列。
在另一態樣中,提供一種經分離聚核苷酸,其編碼包含編碼在4-1BB處特異性結合的抗體或抗體片段之序列的經分離聚核苷酸。
在某些實施例中,該聚核苷酸或核酸為DNA。在其他實施例中,本發明之聚核苷酸為RNA,例如呈信使RNA (mRNA)形式。本發明之RNA可為單股或雙股RNA。
重組方法本發明之雙特異性抗體可例如藉由固態肽合成(例如梅里菲爾德固相合成(Merrifield solid phase synthesis))或重組製造獲得。對於重組製造,例如上文所述之一或多個編碼雙特異性抗原結合分子或其多肽片段之聚核苷酸經分離且插入至一或多個載體中以供在宿主細胞中進一步選殖及/或表現。此類聚核苷酸可使用習知程序容易地分離及測序。在本發明之一個態樣中,提供包含本發明之聚核苷酸中之一或多者的載體,較佳表現載體。可使用熟習此項技術者熟知的方法構築含有活化T細胞之雙特異性抗原結合分子(片段)之編碼序列與適當轉錄/轉譯控制信號的表現載體。此等方法包括活體外重組DNA技術、合成技術及活體內重組/基因重組。參看例如Maniatis等人, MOLECULAR CLONING: A LABORATORY MANUAL, Cold Spring Harbor Laboratory, N.Y. (1989);及Ausubel等人, CURRENT PROTOCOLS IN MOLECULAR BIOLOGY, Greene Publishing Associates and Wiley Interscience, N.Y. (1989)中所描述之技術。表現載體可為質體、病毒之一部分,或可為核酸片段。表現載體包括表現卡匣,其中編碼雙特異性抗原結合分子或其多肽片段之聚核苷酸(即編碼區)以與啟動子及/或其他轉錄或轉譯控制元件可操作結合之方式選殖。如本文所用,「編碼區」為由轉譯成胺基酸之密碼子組成的核酸之一部分。「終止密碼子」(TAG、TGA或TAA)雖然未轉譯成胺基酸,但其可視為編碼區(若存在)之一部分,但任何側接序列(例如啟動子、核糖體結合位點、轉錄終止子、內含子、5'及3'未轉譯區及類似序列)並非編碼區之一部分。兩個或更多個編碼區可存在於單個聚核苷酸構築體中,例如單個載體上,或存在於各別聚核苷酸構築體中,例如各別(不同)載體上。此外,任何載體可含有單個編碼區,或可包含兩個或更多個編碼區,例如本發明之載體可編碼一或多種多肽,其經蛋白水解分裂而轉譯後或共轉譯分離成最終蛋白質。另外,本發明之載體、聚核苷酸或核酸可編碼異源編碼區,其融合或未融合至編碼本發明之雙特異性抗原結合分子或其多肽片段的聚核苷酸,或其變異體或衍生物。異質編碼區包括(不限於)專門化元件或基元,諸如分泌性信號肽或異質功能域。可操作結合為例如多肽之基因產物的編碼區與一或多個調節序列按使基因產物之表現受到調節序列之影響或控制的方式結合。若誘導啟動子功能導致編碼所要基因產物之mRNA轉錄且若兩個DNA片段之間連接的性質不干擾表現調節序列導引基因產物表現的能力或不干擾DNA模板轉錄的能力,則兩個DNA片段(諸如多肽編碼區及與其相關聯的啟動子)為「可操作地相關聯」。因此,若啟動子能夠實現編碼多肽之核酸的轉錄,則啟動子區域將與彼核酸可操作地相關聯。啟動子可為僅導引預定細胞中之DNA實質性轉錄之細胞特異性啟動子。除啟動子以外的其他轉錄控制元件(例如增強子、操縱子、抑制子及轉錄終止信號)與引導細胞特異性轉錄之聚核苷酸可操作地結合。
適合啟動子及其他轉錄控制區揭示於本文中。多種轉錄控制區已為熟習此項技術者所知。此等區域包括(但不限於)在脊椎動物細胞中起作用的轉錄控制區,諸如(但不限於)啟動子及強化子區段,其來自巨細胞病毒(例如即刻早期啟動子,連同內含子A)、猿猴病毒40 (例如早期啟動子)及反轉錄病毒(諸如勞斯肉瘤病毒(Rous sarcoma virus))。其他轉錄控制區包括來源於脊椎動物基因(諸如肌動蛋白、熱休克蛋白、牛生長激素及兔α-血球蛋白)之區域,以及能夠控制真核細胞中之基因表現的其他序列。額外適合的轉錄控制區包括組織特異性啟動子及增強子以及誘導性啟動子(例如啟動子誘導性四環素(tetracyclins))。類似地,多種轉譯控制元件已為一般技術者所知。此等元件包括(但不限於)核糖體結合位點、轉譯起始及終止密碼子,以及來源於病毒系統的元件(特定言之,內部核糖體入口位點或IRES,亦稱為CITE序列)。表現卡匣亦可包括其他特徵,諸如複製起點,及/或染色體整合元件,諸如反轉錄病毒長末端重複序列(LTR),或腺相關病毒(AAV)反向末端重複序列(ITR)。
本發明之聚核苷酸及核酸編碼區可與編碼分泌肽或信號肽的其他編碼區結合,從而引導由本發明之聚核苷酸編碼的多肽的分泌。舉例而言,若需要分泌雙特異性抗原結合分子或其多肽片段,則編碼信號序列之DNA可置於編碼本發明之雙特異性抗原結合分子的核酸或其多肽片段上游。根據信號假設,哺乳動物細胞所分泌的蛋白質具有自成熟蛋白質裂解(一旦生長蛋白質鏈跨越粗糙內質網輸出已起始)的信號肽或分泌性前導序列。一般熟習此項技術者認識到,脊椎動物細胞分泌的多肽通常具有與多肽之N端融合的信號肽,該信號肽自所轉譯之多肽裂解而產生呈分泌或「成熟」形式的多肽。在某些實施例中,使用原生信號肽(例如免疫球蛋白重鏈或輕鏈信號肽),或該序列之功能衍生物,該功能衍生物保留引導與其可操作地結合之多肽之分泌的能力。或者,可使用異源哺乳動物信號肽或其功能衍生物。舉例而言,野生型前導序列可經人類組織纖維蛋白溶酶原活化因子(TPA)或小鼠β-葡糖醛酸酶之前導序列取代。
編碼可用於促進隨後純化(例如組胺酸標籤)或幫助標記融合蛋白之短蛋白質序列之DNA可包括於編碼本發明之雙特異性抗原結合分子的聚核苷酸或其多肽片段內或末端處。
在本發明之另一態樣中,提供包含一或多種本發明之聚核苷酸的宿主細胞。在某些態樣中,提供包含一或多種本發明之載體的宿主細胞。聚核苷酸及載體可合併本文分別關於聚核苷酸及載體所述之任一特徵(單個或組合)。在一個態樣中,宿主細胞包含載體(例如經載體轉型或轉染),該載體包含編碼本發明之雙特異性抗原結合分子的聚核苷酸(或其部分)。如本文中所使用,術語「宿主細胞」係指任何類型之可經工程改造以產生本發明之融合蛋白質或其片段的細胞系統。適用於複製及支持抗原結合分子之表現的宿主細胞為此項技術中熟知。此類細胞可視需要經特定表現載體轉染或轉導,且可生長含有大量載體之細胞以用於接種大型醱酵槽,以獲得足量抗原結合分子以用於臨床應用。適合的宿主細胞包括原核微生物,諸如大腸桿菌,或各種真核生物細胞,諸如中國倉鼠卵巢細胞(CHO)、昆蟲細胞或其類似物。舉例而言,可利用細菌產生多肽,特別是不需要糖基化時。在表現之後,可自可溶性部分之細菌細胞糊狀物分離出多肽且可將該多肽進一步純化。除原核生物外,諸如絲狀真菌或酵母之真核微生物亦為適用於編碼抗體之載體的選殖或表現宿主,包括糖基化路徑已經「人類化」、從而產生具有部分或完全人類糖基化型態之多肽的真菌及酵母株。參看Gerngross, Nat Biotech 22, 1409-1414 (2004),及Li等人, Nat Biotech 24, 210-215 (2006)。
適用於表現(糖基化)多肽的宿主細胞亦來源於多細胞生物體(無脊椎動物及脊椎動物)。無脊椎動物細胞之實例包括植物及昆蟲細胞。已鑑別出眾多可與昆蟲細胞聯合使用,尤其用於轉染草地黏蟲(Spodoptera frugiperda)細胞之桿狀病毒株。植物細胞培養物亦可用作宿主。參看例如美國專利第5,959,177號、第6,040,498號、第6,420,548號、第7,125,978號及第6,417,429號(描述利用轉殖基因植物產生抗體的PLANTIBODIES
TM技術)。脊椎動物細胞亦可用作宿主。舉例而言,適於在懸浮液中生長之哺乳動物細胞株可為適用的。適用哺乳動物宿主細胞株之其他實例為經SV40轉型的猴腎CV1細胞株(COS-7);人類胎腎細胞株(293或293T細胞,如例如Graham等人, J Gen Virol 36, 59 (1977)中所述);幼倉鼠腎細胞(BHK);小鼠塞特利氏細胞(mouse sertoli cell) (TM4細胞,如例如Mather, Biol Reprod 23, 243-251 (1980)中所述);猴腎細胞(CV1);非洲綠猴腎細胞(VERO-76);人類子宮頸癌細胞(HELA);犬腎細胞(MDCK);水牛鼠肝細胞(buffalo rat liver cell) (BRL 3A);人類肺細胞(W138);人類肝細胞(Hep G2);小鼠乳腺腫瘤細胞(MMT 060562);TRI細胞(如例如Mather等人, Annals N. Y. Acad Sci 383, 44-68 (1982)中所述);MRC 5細胞及FS4細胞。其他適用的哺乳動物宿主細胞株包括中國倉鼠卵巢(CHO)細胞,包括dhfr
-CHO細胞(Urlaub等人, Proc Natl Acad Sci USA 77, 4216 (1980));及骨髓瘤細胞株,諸如YO、NS0、P3X63及Sp2/0。關於適於產生蛋白質的某些哺乳動物宿主細胞株之綜述,參看例如Yazaki及Wu, Methods in Molecular Biology, 第248卷(B.K.C. Lo編, Humana Press, Totowa, NJ), 第255-268頁 (2003)。宿主細胞包括經培養細胞,例如經培養之哺乳動物細胞、酵母細胞、昆蟲細胞、細菌細胞及植物細胞(僅舉數例),而且包括轉殖基因動物、轉殖基因植物或經培養之植物或動物組織中所包含的細胞。在一個實施例中,宿主細胞為真核生物細胞,較佳為哺乳動物細胞,諸如中國倉鼠卵巢(CHO)細胞、人類胚腎(HEK)細胞或淋巴細胞(例如Y0、NS0、Sp20細胞)。此項技術中已知在此等系統中表現外源基因的標準技術。表現包含免疫球蛋白之重鏈或輕鏈之多肽的細胞可經工程改造以亦表現免疫球蛋白鏈中之另一者,使得所表現產物為具有重鏈及輕鏈的免疫球蛋白。
在一個態樣中,提供製造本發明之雙特異性抗原結合分子或其多肽片段的方法,其中該方法包含在適於表現本發明之雙特異性抗原結合分子或其多肽片段的條件下培養如本文所提供的包含編碼本發明之雙特異性抗原結合分子的聚核苷酸或其多肽片段的宿主細胞,且自宿主細胞(或宿主細胞培養基)回收本發明之雙特異性抗原結合分子或其多肽片段。
如本文所述製備的本發明之雙特異性分子可藉由此項技術中已知的技術(諸如高效液相層析法、離子交換層析法、凝膠電泳、親和力層析法、尺寸排阻層析法等)純化。用於純化特定蛋白質之實際條件部分地視諸如淨電荷、疏水性、親水性等因素而定,且對於熟習此項技術者而言為顯而易見的。對於親和力層析純化,可使用雙特異性抗原結合分子結合之抗體、配位體、受體或抗原。舉例而言,關於本發明之融合蛋白質之親和層析純化,可使用具有蛋白質A或蛋白質G之基質。可使用序列蛋白質A或G親和層析及尺寸排阻層析來分離實質上如實例中所描述之抗原結合分子。雙特異性抗原結合分子或其片段之純度可藉由多種眾所周知的分析方法中之任一者測定,包括凝膠電泳、高壓液相層析法等。舉例而言,如實例中所述表現之雙特異性抗原結合分子顯示為完整的且如還原及非還原SDS-PAGE所證實適當地裝配。
分析可藉由此項技術中已知之各種分析對本文中提供之抗原結合分子針對其物理/化學特性及/或生物活性進行鑑別、篩選或表徵。
1.
親和力分析本文提供之雙特異性抗原結合分子、抗體及抗體片段對相應TNF受體之親和力可根據實例中所述之方法,藉由表面電漿子共振(SPR),使用諸如BIAcore儀器(GE Healthcare)之標準儀器測定,且受體或目標蛋白諸如可藉由重組表現獲得。雙特異性抗原結合分子對目標細胞抗原之親和力亦可藉由表面電漿子共振(SPR),使用諸如BIAcore儀器(GE Healthcare)之標準儀器測定,且受體或目標蛋白諸如可藉由重組表現獲得。用於量測結合親和力之特定說明性及例示性實施例描述於實例2中。根據一個態樣,藉由表面電漿子共振、使用BIACORE® T100機器(GE Healthcare)、在25℃量測K
D。
2. 結合分析及其他分析本文提供之雙特異性抗原結合分子與表現相應受體之細胞的結合可例如藉由流動式細胞量測術(FACS)使用表現具體受體或目標抗原之細胞株評估。在一個態樣中,結合分析中使用表現TNF受體之新鮮的周邊血液單核細胞(PBMC)。此等細胞在分離後(原生PMBC)或在刺激後(經活化之PMBC)直接使用。在另一態樣中,使用經活化之小鼠脾細胞(表現TNF受體分子)證明本發明之雙特異性抗原結合分子或抗體與表現相應TNF受體之細胞的結合。在另一態樣中,自健康食蟹獼猴之肝素化血液分離之PBMC用於顯示相應表現食蟹獼猴TNF受體之細胞的雙特異性抗原結合分子或抗體。
在另一態樣中,使用表現目標細胞抗原(例如FAP)之癌細胞株說明抗原結合分子與目標細胞抗原之結合。
在另一態樣中,可使用競爭分析來鑑別分別與特定抗體或抗原結合分子競爭結合於目標或TNF受體之抗原結合分子。在某些實施例中,此類競爭抗原結合分子結合於與由特異性抗目標抗體或特異性抗TNF受體抗體所結合相同之抗原決定基(例如直鏈或構形抗原決定基)。用於定位抗體所結合之抗原決定基之具體例示性方法提供於Morris (1996) 「Epitope Mapping Protocols」, Methods in Molecular Biology 第66卷 (Humana Press, Totowa, NJ)中。
3. 活性分析在一個態樣中,提供用於鑑別雙特異性抗原結合分子之分析法,該等分子結合於特異性目標細胞抗原及具有生物活性的特異性TNF受體。生物活性可包括例如經細胞上表現目標細胞抗原之TNF受體進行之促效性信號傳導。亦提供由分析鑑別為具有此類活體外生物活性之含有TNF家族配位三聚體之抗原結合分子。
在某些態樣中,測試本發明之雙特異性抗原結合分子的此類生物活性。此外,用於偵測細胞溶解(例如藉由量測LDH釋放)、誘導之細胞凋亡動力學(例如藉由量測卡斯蛋白酶3/7活性)或細胞凋亡(例如使用TUNEL分析)之分析為此項技術中熟知的。此外,此類複合物之生物活性可藉由評估其對多種淋巴細胞子集(諸如NK細胞、NKT細胞或γδ T細胞)之存活、增殖及淋巴激素分泌之作用或評估其調節抗原呈現細胞(諸如樹突狀細胞、單核細胞/巨噬細胞或B細胞)之表現型及功能的能力來評估。
醫藥組合物、調配物及投與途徑在另一態樣中,本發明提供包含本文所提供雙特異性抗原結合分子或抗體中之任一者的醫藥組合物,其例如用於以下治療方法中之任一者中。在一個實施例中,醫藥組合物包含本文提供的雙特異性抗原結合分子中之任一者及至少一種醫藥學上可接受之賦形劑。在另一實施例中,醫藥組合物包含本文提供之雙特異性抗原結合分子中之任一者及至少一種例如下文所述之額外治療劑。
本發明的醫藥組合物包含治療有效量之一或多種雙特異性抗原結合分子,其溶解或分散於醫藥學上可接受之賦形劑中。短語「醫藥學上或藥理學上可接受」係指分子實體及組合物在所用劑量及濃度下對於接受者而言一般為無毒性的,亦即當適當時投與動物(諸如人類)時,不會產生有害的過敏反應或其他不良反應。含有至少一種根據本發明之雙特異性抗原結合分子或抗體及視情況選用之額外活性成分的醫藥組合物的製備將為熟習本發明之技術者已知,如以引用的方式併入本文中Remington's Pharmaceutical Sciences, 第18版. Mack Printing Company, 1990所例示。特定言之,組合物為凍乾調配物或水溶液。如本文所用,如一般技術人員已知,「醫藥學上可接受之賦形劑」包括任何及全部溶劑、緩衝液、分散液培養基、塗料、界面活性劑、抗氧化劑、防腐劑(例如抗菌劑、抗真菌劑)、等張劑、鹽、穩定劑以及其組合。
非經腸組合物包括經設計以藉由注射(例如皮下、皮內、病灶內、靜脈內、動脈內、肌肉內、鞘內或腹膜內注射)來投與的組合物。對於注射,本發明之雙特異性抗原結合分子或抗體可在水溶液中,較佳生理學相容緩衝液(諸如漢克氏溶液(Hanks'solution)、林格氏溶液(Ringer's solution)或生理食鹽水緩衝液)中調配。溶液可含有調配劑,諸如懸浮劑、穩定劑及/或分散劑。或者,雙特異性抗原結合分子或抗體可在使用之前呈用適合媒劑(例如無菌無熱原質水)復原之粉末形式。無菌可注射溶液藉由將所需量的本發明之抗原結合分子視需要與下文列舉的多種其他成分一起併入適當溶劑中來製備。無菌性可容易藉由例如經由無菌過濾膜過濾來實現。一般而言,分散液係藉由將各種經滅菌之活性成分併入含有基本分散介質及/或其他成分的無菌媒劑中來製備。在用於製備無菌可注射溶液之無菌粉末情況下,較佳製備方法為真空乾燥及冷凍乾燥技術,其利用預先無菌過濾之液體介質產生活性成分與任何其他所需成分之粉末。必要時,液體介質宜經緩衝,且在注射之前,首先用足量鹽水或葡萄糖使液體稀釋劑呈等張性。該組合物在製造及儲存條件下必須為穩定的,且必須避免諸如細菌及真菌之微生物的污染作用。應瞭解,內毒素污染應最低限度地保持在安全水準,例如低於0.5 ng/mg蛋白質。適合的醫藥學上可接受之賦形劑包括(但不限於):緩衝液,諸如磷酸鹽、檸檬酸鹽及其他有機酸;抗氧化劑,包括抗壞血酸及甲硫胺酸;防腐劑(諸如十八烷基二甲基苯甲基氯化銨;氯化六羥季銨;苯紮氯銨;苄索氯銨;酚、丁醇或苯甲醇;對羥基苯甲酸烷基酯,諸如對羥基苯甲酸甲酯或對羥基苯甲酸丙酯;兒茶酚;間苯二酚;環己醇;3-戊醇;及間甲酚);低分子量(小於約10個殘基)多肽;蛋白質,諸如血清白蛋白、明膠或免疫球蛋白;親水性聚合物,諸如聚乙烯吡咯啶酮;胺基酸,諸如甘胺酸、麩醯胺酸、天冬醯胺、組胺酸、精胺酸或離胺酸;單醣、雙醣及其他碳水化合物,包括葡萄糖、甘露糖或糊精;螯合劑,諸如EDTA;糖,諸如蔗糖、甘露糖醇、海藻糖或山梨糖醇;成鹽相對離子,諸如鈉;金屬錯合物(例如Zn-蛋白質錯合物);及/或非離子性界面活性劑,諸如聚乙二醇(PEG)。水性注射懸浮液可含有增加懸浮液黏度之化合物,諸如羧甲基纖維素鈉、山梨糖醇、聚葡萄糖或其類似物。視情況,懸浮液亦可含有適合穩定劑或增加化合物溶解性之試劑以允許製備高度濃縮之溶液。此外,活性化合物之懸浮液可製備成適當的油性注射懸浮液。適合親脂性溶劑或媒劑包括脂肪油,諸如芝麻油;或合成脂肪酸酯,諸如油酸乙酯或甘油三酯;或脂質體。
活性成分可截留於微膠囊中,例如藉由凝聚技術或藉由界面聚合法所製備之微膠囊,例如分別為羥基甲基纖維素或明膠微膠囊及聚(甲基丙烯酸甲酯)微膠囊;截留於膠態藥物遞送系統(例如脂質體、白蛋白微球體、微乳液、奈米顆粒及奈米膠囊)中或巨乳液中。此類技術揭示於Remington's Pharmaceutical Sciences (第18版, Mack Printing Company, 1990)中。可製備持續釋放製劑。持續釋放型製劑之適合實例包括含有多肽之固體疏水性聚合物之半滲透基質,該等基質呈成形物品形式,例如膜或微膠囊。在特定實施例中,可注射組合物之延長吸收可藉由在組合物中使用延遲吸收劑(諸如單硬脂酸鋁、明膠或其組合)來達成。
本文中之例示性醫藥學上可接受之載劑進一步包括間質藥物分散劑,諸如可溶性中性活性玻尿酸酶糖蛋白(sHASEGP),例如人類可溶性PH-20玻尿酸酶糖蛋白,諸如rHuPH20 (HYLENEX®, Baxter International, Inc.)。某些示例性sHASEGP及使用方法,包括rHuPH20,描述於美國專利公開案第2005/0260186號及第2006/0104968號中。在一個態樣中,sHASEGP與一或多種其他葡萄糖胺聚糖酶(諸如軟骨素酶)組合。
例示性凍乾抗體調配物描述於美國專利第6,267,958號中。抗體調配物水溶液包括美國專利第6,171,586號及WO2006/044908中所描述之抗體調配物水溶液,WO2006/044908中所描述之調配物包括組胺酸-乙酸鹽緩衝液。
除了事先描述的組合物之外,抗原結合分子亦可調配為儲槽製劑。此類長效調配物可藉由植入(例如皮下或肌肉內植入)或藉由肌肉內注射來投與。因此,舉例而言,融合蛋白質可用適合的聚合或疏水性材料(例如呈可接受之油中的乳液形式)或離子交換樹脂調配,或調配成微溶性衍生物,例如微溶性鹽。
包含本發明之雙特異性抗原結合分子或抗體的醫藥組合物可以藉助於習知混合、溶解、乳化、囊封、包覆或凍乾法製造。醫藥組合物可使用一或多種有利於將蛋白質處理成可在醫藥學上使用之製劑的生理學上可接受之載劑、稀釋劑、賦形劑或助劑以習知方式調配。適當調配物視所選投與途徑而定。
雙特異性抗原結合分子可以調配成游離酸或鹼、中性或鹽形式之組合物。醫藥學上可接受之鹽為實質上保留游離酸或鹼之生物活性的鹽。此等鹽包括酸加成鹽,例如與蛋白質組合物之游離胺基形成的鹽,或與無機酸(諸如鹽酸或磷酸)或有機酸(諸如乙酸、乙二酸、酒石酸或杏仁酸)形成的鹽。與游離羧基形成的鹽亦可衍生自無機鹼,諸如氫氧化鈉、氫氧化鉀、氫氧化銨、氫氧化鈣或氫氧化鐵;或有機鹼,諸如異丙胺、三甲胺、組胺酸或普魯卡因(procaine)。相較於對應游離鹼形式,醫藥鹽傾向於更溶於水性及其他質子溶劑中。
本文中之組合物亦可含有一種以上為所治療之特定適應症所必需之活性成分,較佳為具有不會對彼此產生不利影響之互補活性的活性成分。此類活性成分適合地以對預期目的有效之量組合存在。
用於活體內投與之調配物通常為無菌的。無菌性可容易藉由例如經由無菌過濾膜過濾來實現。
治療方法及組合物本文所提供之雙特異性抗原結合分子或抗體中之任一者可用於治療方法中。
為了用於治療方法中,本發明之雙特異性抗原結合分子或抗體可用與良好醫學實踐相符之方式調配、給藥及投與。在此情形中考慮之因素包括所治療之特定病症、所治療之特定哺乳動物、個別患者之臨床病況、病症起因、藥劑遞送部位、投與方法、投與時程及醫學從業者已知的其他因素。
在一個態樣中,提供本發明之雙特異性抗原結合分子或抗體用作藥物。
在其他態樣中,提供本發明之雙特異性抗原結合分子或抗體,其用於(i)刺激或提高T細胞反應,(ii)支持經活化之T細胞存活,(iii)治療感染,(iv)治療癌症,(v)延遲癌症進展,或(vi)延長罹患癌症之患者的存活。在一個特定態樣中,提供本發明的含有TNF家族配位體三聚物之抗原結合分子或抗體,其用於治療疾病,特定言之用於治療癌症。
在某些態樣中,提供本發明之雙特異性抗原結合分子或抗體,其用於治療方法中。在一個態樣中,本發明提供如本文所述之雙特異性抗原結合分子或抗體,其用於治療有需要之個體的疾病。在某些態樣中,本發明提供一種雙特異性抗原結合分子或抗體,其用於治療患有疾病之個體的方法中,該方法包含向個別投與治療有效量之雙特異性抗原結合分子或抗體。在某些態樣中,待治療之疾病為癌症。個體、患者或需要治療之「個體」典型地為哺乳動物,更特定言之,人類。
在一個態樣中,提供一種方法,其用於(i)刺激或提高T細胞反應,(ii)支持經活化之T細胞存活,(iii)治療感染,(iv)治療癌症,(v)延遲癌症進展或(vi)延長罹患癌症之患者的存活,其中該方法包含向有需要之個體投與治療有效量的本發明之雙特異性抗原結合分子或抗體。
在另一態樣中,本發明提供本發明之雙特異性抗原結合分子或抗體的用途,其用於製造或製備供治療有需要之個體中的疾病的藥劑。在一個態樣中,藥劑用於治療疾病之方法,其包含投與患有疾病之個體治療有效量之藥劑。在某些態樣中,所治療之疾病為增生性病症,特定言之,癌症。癌症的實例包括(但不限於)膀胱癌、腦癌、頭頸癌、胰臟癌、肺癌、乳癌、卵巢癌、子宮癌、子宮頸癌、子宮內膜癌、食道癌、結腸癌、結腸直腸癌、直腸癌、胃癌、前列腺癌、血液癌、皮膚癌、鱗狀細胞癌、骨癌及腎癌。癌症之其他實例包括癌瘤、淋巴瘤(例如霍奇金氏及非霍奇金氏淋巴瘤)、母細胞瘤、肉瘤及白血病。可使用本發明之雙特異性抗原結合分子或抗體治療的其他細胞增殖病症包括(但不限於)定位於以下部位中之贅瘤:腹部、骨骼、乳房、消化系統、肝臟、胰臟、腹膜、內分泌腺體(腎上腺、副甲狀腺、垂體、睾丸、卵巢、胸腺、甲狀腺)、眼部、頭部及頸部、神經系統(中樞及周邊)、淋巴系統、骨盆、皮膚、軟組織、脾臟、胸部以及泌尿生殖系統。亦包括癌前病況或病變及癌症轉移。在某些實施例中,癌症係選自由以下組成之群:腎細胞癌、皮膚癌、肺癌、結腸直腸癌、乳癌、腦癌、頭頸癌。熟習此項技術者容易認識到在多數情況下,本發明之雙特異性抗原結合分子或抗體可能不提供治癒但可提供益處。在一些實施例中,具有一些益處之生理學變化亦視為治療上有利的。因此,在一些態樣中,提供生理學改變的本發明之雙特異性抗原結合分子或抗體之量視為「有效量」或「治療有效量」。
為了預防或治療疾病,本發明之雙特異性抗原結合分子或抗體(當單獨使用或與一或多種其他額外治療劑組合使用時)的適當劑量將取決於待治療的疾病類型、投與途徑、患者體重、特異性分子、疾病之嚴重程度及時程、本發明之雙特異性抗原結合分子或抗體是出於預防性還是治療性目的投與、先前或同時治療性干預、患者之臨床病史及對融合蛋白之反應,以及主治醫師之判斷。負責投與的從業者將在任何情況下確定組合物中活性成分之濃度及適用於單獨個體的劑量。本文中涵蓋各種給藥時程,包括(但不限於)單次投與或經各個時間點多次投與、快速投與及脈衝式輸注。
本發明之雙特異性抗原結合分子或抗體適合一次性或經一連串治療向患者投與。視疾病之類型及嚴重程度而定,約1 µg/kg至15 mg/kg (例如0.1 mg/kg-10 mg/kg)之含有TNF家族配位體三聚物之抗原結合分子可為投與患者之初始候選劑量,無論例如藉由一或多次單獨投與或藉由連續輸注。視上文所述之因素而定,一個典型日劑量可在約1 μg/kg至100 mg/kg或更多之範圍內。對於歷經數日或更長時間之重複投與,治療一般將視病況而定持續至發生疾病症狀之所需抑制為止。本發明之雙特異性抗原結合分子或抗體的一個例示性劑量將在約0.005 mg/kg至約10 mg/kg範圍內。在其他實例中,劑量亦可包含每次投與約1 μg/kg體重、約5 μg/kg體重、約10 μg/kg體重、約50 μg/kg體重、約100 μg/kg體重、約200 μg/kg體重、約350 μg/kg體重、約500 μg/kg體重、約1 mg/kg體重、約5 mg/kg體重、約10 mg/kg體重、約50 mg/kg體重、約100 mg/kg體重、約200 mg/kg體重、約350 mg/kg體重、約500 mg/kg體重至約1000 mg/kg體重或更高,及其中可衍生之任何範圍。在來自本文中列舉之數值之可衍生範圍之實例中,基於上述數值,可投與約5 mg/kg體重至約100 mg/kg體重、約5 μg/kg體重至約500 mg/kg體重等之範圍。因此,可向患者投與約0.5 mg/kg、2.0 mg/kg、5.0 mg/kg或10 mg/kg (或其任何組合)之一或多種劑量。此類劑量可間歇地投與,例如每週或每三週(例如以使得患者接收約二至約二十或例如約六個劑量之融合蛋白)。可投與初始較高起始劑量,隨後可投與一或多種較低劑量。然而,其他給藥方案可為適用的。此療法之進展易於藉由習知技術及分析監測。
本發明之雙特異性抗原結合分子或抗體一般將以有效實現預期目的之量使用。為了用於治療或預防疾病病狀,本發明之雙特異性抗原結合分子或抗體或其醫藥組合物以治療有效量投與或施用。治療有效量之確定完全屬於熟習此項技術者之能力範圍內,尤其根據本文所提供之詳細揭示內容。
關於全身性投與,可首先自活體外分析(諸如細胞培養分析)估算治療有效劑量。接著可在動物模型中調配劑量以實現循環濃度範圍,包括如在細胞培養物中測定的IC50。此類資訊可用於更準確地判定適用於人類之劑量。
初始劑量亦可自活體內資料(例如動物模型)、使用此項技術中熟知的技術估算。一般熟習此項技術者容易基於動物資料最佳化人類投與。
可以獨立地調整劑量及間隔以提供足以維持治療作用的本發明之雙特異性抗原結合分子或抗體的血漿含量。常見的患者注射投與劑量範圍為約0.1至50毫克/公斤/天,通常為約0.5至1毫克/公斤/天。治療有效的血漿含量可藉由每日投與多次劑量來達成。血漿含量可藉由例如HPLC量測。
在局部投與或選擇性攝入之情況下,本發明之雙特異性抗原結合分子或抗體的有效局部濃度可能與血漿濃度無關。熟習此項技術者能夠最佳化治療有效局部劑量而無需不當實驗。
本文所述的本發明之雙特異性抗原結合分子或抗體的治療有效劑量一般將提供治療益處而不引起實質毒性。可在細胞培養物或實驗動物中藉由標準醫藥學程序測定融合蛋白質之毒性及治療功效。可使用細胞培養分析及動物研究來確定LD
50(50%群體致死劑量)及ED
50(50%群體治療有效劑量)。毒性作用與治療作用之間的劑量比率為治療指數,其可表示為比率LD
50/ED
50。較佳為展現大治療指數的雙特異性抗原結合分子。在一個態樣中,本發明之雙特異性抗原結合分子或抗體展現高治療指數。自細胞培養分析及動物研究獲得之資料可用於調配適用於人類的劑量範圍。該劑量較佳在循環濃度範圍內,包括毒性極小或無毒性的ED50。該劑量可視各種因素而定在此範圍內變化,該等因素例如所採用之劑型、所採用之投與途徑、個體之病狀及類似因素。精確配方、投與途徑及劑量可由個別醫師根據患者之病狀選擇(參看例如Fingl等人, 1975, in: The Pharmacological Basis of Therapeutics, 第1章, 第1頁, 以全文引用的方式併入本文中)。
使用本發明融合蛋白質治療患者之主治醫師將知曉如何及何時由於毒性、器官功能不全及其類似原因而終止、中斷或調整投藥。相反,主治醫師亦知曉若臨床反應不充足(排除毒性),則將治療調整至較高水準。處理所關注病症時所投與劑量之量值將因所治療病狀之嚴重程度及投與途徑及類似因素而不同。病況之嚴重程度可例如部分地藉由標準預後評估方法來評估。另外,劑量及可能的給藥頻率亦將根據個別患者之年齡、體重及反應而變化。
其他藥劑及治療本發明之雙特異性抗原結合分子或抗體可在療法中與一或多種其他藥劑組合投與。舉例而言,本發明之雙特異性抗原結合分子或抗體可與至少一種額外治療劑共投與。術語「治療劑」涵蓋可投與用於治療需要此類治療之個體中之症狀或疾病的任何藥劑。此類其他治療劑可包含適於所治療特定適應症的任何活性成分,較佳為彼此間具有互補活性、無不利影響的彼等活性成分。在某些實施例中,額外治療劑為另一抗癌劑,例如微管瓦解劑、抗代謝物、拓撲異構酶抑制劑(topoisomerase inhibitor)、DNA嵌入劑、烷基化劑、激素療法、激酶抑制劑、受體拮抗劑、腫瘤細胞凋亡活化劑或抗血管生成劑。在某些態樣中,額外治療劑為免疫調節劑、細胞抑制劑、細胞黏附抑制劑、細胞毒性或細胞生長抑制劑、細胞凋亡活化劑或提高細胞對細胞凋亡誘導劑之靈敏度的藥劑。
此類其他藥劑宜以可有效達成預定目的之量組合存在。此類其他藥劑之有效量視所使用之融合蛋白質之量、病症或治療之類型及如上文所述之其他因素而定。本發明之雙特異性抗原結合分子或抗體一般以如本文所述之相同劑量及投與途徑,或約1至99%之本文所述劑量,或經驗上/臨床上確定為適當的任何劑量及任何途徑使用。
上文提及之此類組合療法涵蓋組合投藥(其中在相同或不同組合物中包括兩種或更多種治療劑)以及分開投藥,在此情況下,本發明之雙特異性抗原結合分子或抗體可在投與額外治療劑及/或助劑之前、同時及/或之後進行投藥。
製品在本發明之另一態樣中,提供含有適用於治療、預防及/或診斷上文所述病症之製品。製品包含容器及容器上或容器隨附之標記或藥品說明書。適合的容器包括例如瓶子、小瓶、注射器、IV溶液袋等。容器可由各種材料形成,諸如玻璃或塑膠。容器容納單獨或與有效治療、預防及/或診斷病狀之另一組合物組合之組合物,且可具有無菌接取口(例如容器可為具有可由皮下注射針刺穿之塞子的靜脈內溶液袋或小瓶)。組合物中的至少一種活性劑為本發明之雙特異性抗原結合分子或抗體。
標記或藥品說明書指示組合物用於治療所選病狀。此外,製品可包含(a)其中含有組合物的第一容器,其中組合物包含本發明之T細胞活化雙特異性抗原結合分子;及(b)其中含有組合物的第二容器,其中組合物包含另一種細胞毒性劑或其他治療劑。本發明之此實施例中之製品可進一步包含指示組合物可用於治療特定病狀之藥品說明書。
或者或另外,該製品可進一步包含第二(或第三)容器,其包含醫藥學上可接受之緩衝液,諸如注射用抑菌水(BWFI)、磷酸鹽緩衝鹽水、林格氏溶液(Ringer's solution)及右旋糖溶液。其可進一步包括就商業及使用者觀點而言所需之其他物質,包括其他緩衝液、稀釋劑、過濾器、針及注射器。
表 C ( 序列 ) :
在下文中,列舉本發明之特定實施例:1. 一種雙特異性抗原結合分子,其包含
(a) 至少一個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 至少一個能夠特異性結合於目標細胞抗原之部分體,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
2. 如實施例1之雙特異性抗原結合分子,其中該共刺激TNF受體家族成員選自由OX40及4-1BB組成之群。
3. 如實施例1或2之雙特異性抗原結合分子,其中該共刺激TNF受體家族成員為OX40。
4. 如實施例1至3中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於共刺激TNF受體家族成員之部分體結合於包含胺基酸序列SEQ ID NO:1之多肽。
5. 如實施例1至4中任一項之雙特異性抗原結合分子,其包含至少一個能夠特異性結合於OX40之部分體,其中該部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:2及SEQ ID NO:3組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:4及SEQ ID NO:5組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:6、SEQ ID NO:7、SEQ ID NO:8、SEQ ID NO: 9、SEQ ID NO:10、SEQ ID NO:11及SEQ ID NO:12組成之群的胺基酸序列,
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:13、SEQ ID NO:14及SEQ ID NO:15組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:16、SEQ ID NO:17及SEQ ID NO:18組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:19、SEQ ID NO:20、SEQ ID NO:21、SEQ ID NO:22、SEQ ID NO:23及SEQ ID NO:24組成之群的胺基酸序列。
6. 如實施例1至5中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於OX40之部分體包含重鏈可變區VH,其包含與選自由SEQ ID NO:25、SEQ ID NO:27、SEQ ID NO: 29、SEQ ID NO: 31、SEQ ID NO: 33、SEQ ID NO:35及SEQ ID NO: 37組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區VL,其包含與胺基酸序列SEQ ID NO:26、SEQ ID NO:28、SEQ ID NO: 30、SEQ ID NO: 32、SEQ ID NO: 34、SEQ ID NO:36及SEQ ID NO: 38至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
7. 如實施例1至5中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於OX40之部分體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
8. 如實施例1或2之雙特異性抗原結合分子,其中該共刺激TNF受體家族成員為4-1BB。
9. 如實施例1、2或8中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於共刺激TNF受體家族成員之部分體結合於包含胺基酸序列SEQ ID NO:39之多肽。
10. 如實施例1、2、8或9中任一項之雙特異性抗原結合分子,其包含至少一個能夠特異性結合於4-1BB之部分體,其中該部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:40及SEQ ID NO:41組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:42及SEQ ID NO:43組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47及SEQ ID NO:48組成之群的胺基酸序列
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:49及SEQ ID NO:50組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:51及SEQ ID NO:52組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:53、SEQ ID NO:54、SEQ ID NO:55、SEQ ID NO:56及SEQ ID NO:57組成之群的胺基酸序列。
11. 如實施例1、2、8、9或10中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於4-1BB之部分體包含重鏈可變區VH,其包含與選自由SEQ ID NO:58、SEQ ID NO:60、SEQ ID NO:62、SEQ ID NO:64及SEQ ID NO:66組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與選自由SEQ ID NO:59、SEQ ID NO:61、SEQ ID NO:63、SEQ ID NO:65及SEQ ID NO:67組成之群的胺基酸序列至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
12. 如實施例1、2及8至11中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於4-1BB之部分體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
13. 如實施例1至12中任一項之雙特異性抗原結合分子,其中該目標細胞抗原選自由以下組成之群:纖維母細胞活化蛋白(FAP)、黑素瘤相關硫酸軟骨素蛋白聚糖(MCSP)、表皮生長因子受體(EGFR)、癌胚抗原(CEA)、CD19、CD20及CD33。
14. 如實施例1至13中任一項之雙特異性抗原結合分子,其中該目標細胞抗原為纖維母細胞活化蛋白(FAP)。
15. 如實施例1至13中任一項之雙特異性抗原結合分子,其中該能夠特異性結合於FAP之部分體包含包括以下之VH結構域
(i) CDR-H1,其包含選自由SEQ ID NO:68及SEQ ID NO:69組成之群的胺基酸序列,
(ii) CDR-H2,其包含選自由SEQ ID NO:70及SEQ ID NO:71組成之群的胺基酸序列,及
(iii) CDR-H3,其包含選自由SEQ ID NO:72及SEQ ID NO:73組成之群的胺基酸序列,
及包含以下之VL結構域
(iv) CDR-L1,其包含選自由SEQ ID NO:74及SEQ ID NO:75組成之群的胺基酸序列,
(v) CDR-L2,其包含選自由SEQ ID NO:76及SEQ ID NO:77組成之群的胺基酸序列,及
(vi) CDR-L3,其包含選自由SEQ ID NO:78及SEQ ID NO:79組成之群的胺基酸序列。
16. 如實施例1至7中任一項之雙特異性抗原結合分子,其中
(i) 能夠特異性結合於OX40之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:25、SEQ ID NO: 27、SEQ ID NO:29、SEQ ID NO:31、SEQ ID NO:33、SEQ ID NO:35或SEQ ID NO:37至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:26、SEQ ID NO: 28、SEQ ID NO:30、SEQ ID NO:32、SEQ ID NO:34、SEQ ID NO:36或SEQ ID NO:38至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及
(ii) 能夠特異性結合於FAP之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:80或SEQ ID NO:82至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:81或SEQ ID NO:83至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
17. 如實施例1、2或8至12中任一項之雙特異性抗原結合分子,其中
(i) 能夠特異性結合於4-1BB之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:58、SEQ ID NO: 60、SEQ ID NO:62、SEQ ID NO:64或SEQ ID NO:66至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:59、SEQ ID NO: 61、SEQ ID NO:63、SEQ ID NO:65或SEQ ID NO:67至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及
(ii) 能夠特異性結合於FAP之部分體包含重鏈可變區VH,其包含與胺基酸序列SEQ ID NO:80或SEQ ID NO:82至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列,及輕鏈可變區,其包含與胺基酸序列SEQ ID NO:81或SEQ ID NO:83至少約95%、96%、97%、98%、99%或100%一致的胺基酸序列。
18. 如實施例1至17中任一項之雙特異性抗原結合分子,其中該分子包含
(a) 能夠特異性結合於共刺激TNF受體家族成員之第一Fab片段,
(b) 能夠特異性結合於目標細胞抗原之第二Fab片段,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
19. 如實施例1至18中任一項之雙特異性抗原結合分子,其中該Fc結構域為IgG,特定言之IgG1 Fc結構域或IgG4 Fc結構域。
20. 如實施例1至19中任一項之雙特異性抗原結合分子,其中該Fc結構域包含一或多個降低抗體對Fc受體之結合親和力及/或效應功能的胺基酸取代。
21. 如實施例1至20中任一項之雙特異性抗原結合分子,其中該Fc結構域為具有胺基酸突變L234A、L235A及P329G (根據Kabat EU索引編號)之人類IgG1子類。
22. 如實施例1至21中任一項之雙特異性抗原結合分子,其中該Fc結構域包含促進Fc結構域之第一及第二子單元結合的修飾。
23. 如實施例1至22中任一項之雙特異性抗原結合分子,其中根據杵-臼方法,該Fc結構域之該第一子單元包含杵且該Fc結構域之該第二子單元包含臼。
24. 如實施例1至23中任一項之雙特異性抗體,其中該Fc結構域之該第一子單元包含胺基酸取代S354C及T366W(根據Kabat EU索引編號)且該Fc結構域之該第二子單元包含胺基酸取代Y349C、T366S及Y407V(根據Kabat EU索引編號)。
25. 如實施例1至24中任一項之雙特異性抗原結合分子,其包含
(a) 兩個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 兩個能夠特異性結合於目標細胞抗原之部分體,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
26. 如實施例25之雙特異性抗原結合分子,其中該雙特異性抗原結合分子對於該共刺激TNF受體家族成員及該目標細胞抗原皆為二價。
27. 如實施例1至24中任一項之雙特異性抗原結合分子,其包含(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,以及(b)兩個能夠特異性結合於目標細胞抗原之額外Fab片段,其中該等額外Fab片段各自經肽連接子連接至(a)之重鏈的C端。
28. 如實施例27之雙特異性抗原結合分子,其中該兩個能夠特異性結合於目標細胞抗原之額外Fab片段為交換Fab片段,其中該等可變域VL及VH彼此置換且該等VL-CH鏈各自經肽連接子連接至(a)之重鏈的C端。
29. 如實施例27或28之雙特異性抗原結合分子,其中該兩個能夠特異性結合於共刺激TNF受體家族成員之Fab片段為兩個能夠特異性結合於OX40或4-1BB之Fab片段且該兩個能夠特異性結合於目標細胞抗原之額外Fab片段為能夠特異性結合於FAP之交換Fab片段。
30. 如前述實施例中任一項之雙特異性抗原結合分子,其包含
(a) 兩個能夠特異性結合於共刺激TNF受體家族成員的部分體,
(b) 一個能夠特異性結合於目標細胞抗原之部分體,及
(c) 由能夠穩定結合之第一子單元及第二子單元構成的Fc結構域。
31. 如實施例30之雙特異性抗原結合分子,其中該雙特異性抗原結合分子對於該共刺激TNF受體家族成員為二價且對於該目標細胞抗原為單價。
32. 如前述實施例中任一項之雙特異性抗原結合分子,其包含(a)包含能夠特異性結合於共刺激TNF受體家族成員之兩個Fab片段及Fc結構域的抗體之兩個輕鏈及兩個重鏈,及(b)能夠特異性結合於目標細胞抗原之VH及VL結構域,其中該VH結構域經肽連接子連接至該等重鏈中之一者的C端且其中該VL結構域經肽連接子連接至第二重鏈之C端。
33. 一種聚核苷酸,其編碼如實施例1至32中任一項之雙特異性抗原結合分子。
34. 一種特異性結合於OX40之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL,
(v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL,
(vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或
(vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
35. 一種特異性結合於4-1BB之抗體,其中該抗體包含
(i) 包含胺基酸序列SEQ ID NO:58之重鏈可變區VH及包含胺基酸序列SEQ ID NO:59之輕鏈可變區VL,
(ii) 包含胺基酸序列SEQ ID NO:60之重鏈可變區VH及包含胺基酸序列SEQ ID NO:61之輕鏈可變區VL,
(iii) 包含胺基酸序列SEQ ID NO:62之重鏈可變區VH及包含胺基酸序列SEQ ID NO:63之輕鏈可變區VL,
(iv) 包含胺基酸序列SEQ ID NO:64之重鏈可變區VH及包含胺基酸序列SEQ ID NO:65之輕鏈可變區VL,或
(v) 包含胺基酸序列SEQ ID NO:66之重鏈可變區VH及包含胺基酸序列SEQ ID NO:67之輕鏈可變區VL。
36. 一種聚核苷酸,其編碼如實施例34或35之抗體。
37. 一種醫藥組合物,其包含如實施例1至32中任一項之雙特異性抗原結合分子或如實施例34或35之抗體及至少一種醫藥學上可接受之賦形劑。
38. 如實施例1至32中任一項之雙特異性抗原結合分子,或如實施例34或35之抗體,或如實施例37之醫藥組合物,其用作藥劑。
39. 如1至32實施例中任一項之雙特異性抗原結合分子,或如實施例34或35之抗體,或如實施例37之醫藥組合物,其用於
(i) 刺激T細胞反應,
(ii) 支持活化T細胞之存活,
(iii) 治療感染,
(iv) 治療癌症,
(v) 延遲癌症進展,或
(vi) 延長罹患癌症之患者的存活期。
40. 如實施例1至32中任一項之雙特異性抗原結合分子,或如實施例34或35之抗體,或如实施例37之醫藥組合物,其用於治療癌症。
41. 如實施例1至32中任一項之雙特異性抗原結合分子,或如實施例34或35之抗體,或如實施例37之醫藥組合物,其用於治療癌症,其中該雙特異性抗原結合分子與化學治療劑、輻射及/或其他試劑組合投與用於癌症免疫療法。
42.一種抑制個體中之腫瘤細胞生長的方法,其包含向該個體投與有效量的如實施例1至32中任一項之雙特異性抗原結合分子,或如實施例34或35之抗體,或如實施例37之醫藥組合物,來抑制腫瘤細胞生長。
實例以下為本發明之方法及組合物之實例。應理解,考慮到上文所提供之一般說明,可實施各種其他實施例。
重組型 DNA 技術使用標準方法操縱DNA,如Sambrook等人, Molecular cloning: A laboratory manual; Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, 1989中所述。根據製造商說明書,使用分子生物學試劑。關於人類免疫球蛋白輕鏈及重鏈之核苷酸序列之一般資訊提供於Kabat, E.A.等人, (1991) Sequences of Proteins of Immunological Interest, 第五版, NIH公開案第91-3242號中。
DNA 測序藉由雙股測序法測定DNA序列。
基因合成所需基因區段係使用適當模板藉由PCR產生,或藉由自動化基因合成法,藉由Geneart AG (Regensburg, Germany)自合成寡核苷酸及PCR產物合成。在確切基因序列無法獲得的情況下,根據來自最近同源物的序列來設計寡核苷酸引子且藉由RT-PCR自來源於適當組織之RNA分離基因。將側接有單數個限制性核酸內切酶裂解位點的基因區段選殖至標準選殖/定序載體中。自經轉型之細菌純化質體DNA且藉由UV光譜學來測定濃度。經次選殖之基因片段之DNA序列藉由DNA定序來證實。基因區段經設計具有允許次選殖入各別表現載體中的適合限制位點。所有構築體經設計具有5'端DNA序列,該序列編碼靶向真核細胞中分泌之蛋白質的前導肽。
蛋白質純化參考標準方案,自經過濾之細胞培養物清液層純化蛋白質。簡言之,將抗體施用於蛋白質A瓊脂糖凝膠管柱(GE Healthcare)且用PBS洗滌。在pH 2.8下實現抗體之溶離,接著立即中和樣品。在PBS或20 mm組胺酸、150 mM NaCl pH 6.0中藉由尺寸排阻層析(Superdex 200,GE Healthcare)自單體抗體分離聚集之蛋白質。單體抗體份數經合併、用MILLIPORE Amicon Ultra (30 MWCO)離心濃縮器濃縮(必要時)、凍結且在-20℃或-80℃下儲存。提供部分樣品用於後續蛋白質分析及分析型特徵化,例如藉由SDS-PAGE、尺寸排阻層析(SEC)或質譜。
SDS-PAGE根據製造商說明使用NuPAGE® Pre-Cast凝膠系統(Invitrogen)。特定言之,使用10%或4-12% NuPAGE® Novex® Bis-TRIS Pre-Cast凝膠(pH 6.4)及NuPAGE® MES (還原性凝膠,具有NuPAGE® Antioxidant操作緩衝液添加劑)或MOPS (非還原性凝膠)操作緩衝液。
分析型尺寸排阻層析藉由HPLC層析進行用於測定抗體之聚集及寡聚狀態之尺寸排阻層析(SEC)。簡言之,將蛋白質A純化抗體施用於Agilent HPLC 1100系統上300 mM NaCl、50 mM KH
2PO
4/K
2HPO
4,pH 7.5中之Tosoh TSKgel G3000SW管柱或Dionex HPLC系統上2×PBS中之Superdex 200管柱(GE Healthcare)。藉由UV吸光度及峰值面積之積分來定量溶離之蛋白質。BioRad Gel過濾標準151-1901用作標準。
質譜此部分描述具有VH/VL或CH/CL交換(CrossMabs)之多特異性抗體之表徵,其中強調其正確組裝。藉由去糖基化完整CrossMabs及去糖基化/纖維蛋白溶酶消化或以其他方式去糖基化/受限LysC消化CrossMabs之電噴霧電離質譜(ESI-MS)來分析預期主要結構。
在蛋白質濃度為1 mg/ml下,CrossMab在磷酸鹽或Tris緩衝液中,在37℃下用N-糖苷酶去糖基化17小時。纖維蛋白溶酶或受限LysC (Roche)消化用100 µg去糖基化VH/VL CrossMab分別在Tris緩衝液pH 8中,在室溫下進行120小時及在37℃下進行40分鐘。在質譜分析之前,在Sephadex G25管柱(GE Healthcare)上經由HPLC將樣品去鹽。在配備有TriVersa NanoMate來源(Advion)之maXis 4G UHR-QTOF MS系統(Bruker Daltonik)上經由ESI-MS測定總質量。
實例 1 產生 Ox40 抗體及工具結合子 1.1 抗原之製備、純化及表徵以及藉由噬菌體呈現產生新 OX40 結合子的篩選工具編碼人類、小鼠或獼猴OX40之胞外域的DNA序列(表1)與人類IgG1重鏈CH2及CH3結構域在杵上同框次選殖(Merchant等人, 1998)。在抗原胞外域與人類IgG1之Fc之間引入AcTEV蛋白酶裂解位點。在抗原-Fc杵之C端引入用於引導生物素化之Avi標籤。含有S354C/T366W突變之抗原-Fc杵鏈與含有Y349C/T366S/L368A/Y407V突變之Fc臼鏈之組合允許產生雜二聚體,其包括含有OX40胞外域之鏈之單一複本,因此產生Fc連接之抗原之單體形式(
圖 1A)。
表 1展示多種OX40胞外域之胺基酸序列。
表 2為如圖1中所描繪之單體抗原Fc(kih)融合分子的cDNA及胺基酸序列。
表 1 :抗原胞外域 ( ECD ) 之胺基酸編號及其來源
表 2 : 單體抗原 Fc ( kih ) 融合分子之 cDNA 及胺基酸序列 ( 藉由組合一個 Fc 臼鏈與一個抗原 Fc 杵鏈製備 )
全部OX40-Fc融合編碼序列選殖至驅動來自MPSV啟動子之插入物的表現及含有位於CDS之3'端處的合成聚A信號序列的質體載體。此外,載體含有用於質體之游離型維護之EBV OriP序列。
關於生物素化單體抗原/Fc融合分子之製備,用三個編碼融合蛋白之兩個組分(結及臼鏈)的載體以及BirA (生物素化反應所需之酶)共轉染以指數方式生長之懸浮液HEK293 EBNA細胞。以2:1:0.05比率(「抗原ECD-AcTEV-Fc杵」:「Fc臼」:「BirA」)使用相應載體。
關於在500 ml搖瓶中製造蛋白質,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。關於轉染,使細胞在210 g下離心5分鐘,且用預溫熱之CD CHO培養基置換清液層。使表現載體再懸浮於20 mL含有200 μg載體DNA之CD CHO培養基中。在添加540 μL聚乙烯亞胺(PEI)之後,將溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。在轉染之後一天,向培養物中添加1 mM丙戊酸及7%饋料。培養7天之後,藉由使細胞在210 g下短暫離心15分鐘收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
藉由使用蛋白質A進行親和層析,接著進行尺寸排阻層析自細胞培養物清液層純化所分泌之蛋白質。關於親和力層析,將清液層裝載至經40 mL 20 mM磷酸鈉、20 mM檸檬酸鈉(pH 7.5)平衡的HiTrap蛋白A HP管柱(CV = 5 mL,GE Healthcare)上。藉由用至少10管柱體積之含有20 mM磷酸鈉、20 mM檸檬酸鈉及0.5 M氯化鈉之緩衝液(pH 7.5)洗滌來移除未結合之蛋白質。使用經20管柱體積之20 mM檸檬酸鈉、0.01% (v/v)Tween-20 (pH 3.0)產生之氯化鈉之線性pH梯度(0至500 mM)來溶離結合之蛋白質。管柱接著用10管柱體積的含有20 mM檸檬酸鈉、500 mM氯化鈉及0.01% (v/v) Tween-20 (pH 3.0)的溶液洗滌。
藉由添加1/40 (v/v) 2 M Tris,pH 8.0來調整所收集之溶離份之pH。濃縮且過濾蛋白質,隨後裝載於用2 mM MOPS、150 mM氯化鈉、0.02% (w/v)疊氮化鈉溶液(pH 7.4)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
1.2 自通用 Fab 及常用輕鏈文庫選擇 Ox40 特異性 8H9 、 20B7 、 49B4 、 1G4 、 CLC - 563 、 CLC - 564 及 17A9 抗體抗OX40抗體選自三個不同通用噬菌體呈現文庫:DP88-4 (純系20B7、8H9、1G4及49B4)、共用輕鏈文庫Vk3_20/VH3_23 (純系CLC-563及CLC-564)及λ-DP47 (純系17A9)。
DP88 - 4文庫基於人類生殖系基因使用V結構域對Vk1_5 (κ輕鏈)及VH1_69 (重鏈)建構,其包含輕鏈之CDR3 (L3,3種不同長度)及重鏈之CDR3 (H3,3種不同長度)中的隨機序列間隔。藉由重疊延長(SOE) PCR施加剪接,藉由3 PCR擴增之片段的組合件進行文庫繼代。片段1包含包括隨機L3之抗體基因的5'端,片段2為自L3跨越至H3的中部恆定片段而片段3包含抗體基因之隨機H3及3'部分。分別使用以下引子組合產生DP88-4文庫之此等文庫片段:片段1 (正向引子LMB3與反向引子Vk1_5_L3r_S或Vk1_5_L3r_SY或Vk1_5_L3r_SPY之組合)、片段2 (正向引子RJH31與反向引子RJH32之組合)及片段3 (正向引子DP88-v4-4或DP88-v4-6或DP88-v4-8與反向引子fdseqlong之組合)。用於製造文庫片段之PCR參數為94℃下5分鐘初始變性,1分鐘94℃,1分鐘58℃,1分鐘72℃的25次循環及在72℃下10分鐘的末端延長。對於裝配PCR,使用等莫耳比的凝膠純化之單一片段作為模板,參數為在94℃下3分鐘初始變性及30秒94℃、1分鐘58℃、2分鐘72℃的5次循環。在此階段,添加外部引子(LMB3及fdseqlong)且進行額外20次循環,隨後在72℃下末端延長10分鐘。裝配足夠量的全長隨機Fab構築體之後,將其用NcoI/NheI消化且接合至類似處理的受體噬菌粒載體中。經純化的連接用於約60次轉型至電感受態大腸桿菌TG1中。呈現Fab文庫之噬菌粒粒子經補救且藉由PEG/NaCl純化進行純化,以供選擇。此等文庫建構步驟重複三次以獲得最終文庫尺寸4.4×109。如藉由點漬墨法中之C端標籤偵測所測定的功能性純系之百分比分別為輕鏈92.6%及重鏈93.7%。
共用輕鏈文庫Vk3_20/VH3_23基於人類生殖系基因使用V結構域對Vk3_20 (κ輕鏈)及VH3_23 (重鏈)建構,其包含恆定非隨機共用輕鏈Vk3_20及重鏈之CDR3 (H3,3種不同長度)中的隨機序列間隔。藉由重疊延長(SOE) PCR施加剪接,藉由2 PCR擴增之片段的裝配進行文庫繼代。片段1為自L3跨越至H3的恆定片段,而片段2包含抗體基因之隨機H3及3'部分。分別使用以下引子組合產生Vk3_20/VH3_23共用輕鏈文庫之此等文庫片段:片段1 (正向引子MS64與反向引子DP47CDR3_ba (mod.)之組合)及片段2 (正向引子DP47-v4-4、DP47-v4-6、DP47-v4-8與反向引子fdseqlong之組合)。用於製造文庫片段之PCR參數為94℃下5分鐘初始變性,1分鐘94℃,1分鐘58℃,1分鐘72℃的25次循環及在72℃下10分鐘的末端延長。對於裝配PCR,使用等莫耳比的凝膠純化之單一片段作為模板,參數為在94℃下3分鐘初始變性及30秒94℃、1分鐘58℃、2分鐘72℃的5次循環。在此階段,添加外部引子(MS64及fdseqlong)且進行額外18次循環,隨後在72℃下末端延長10分鐘。裝配足夠量的全長隨機VH構築體之後,將其用MunI/NotI消化且接合至類似處理的受體噬菌粒載體中。經純化的連接用於約60次轉型至電感受態大腸桿菌TG1中。呈現Fab文庫之噬菌粒粒子補救且藉由PEG/NaCl純化進行純化,以供選擇。獲得最終文庫尺寸3.75×109。如藉由點漬墨法中之C端標籤偵測所測定的功能性純系之百分比分別為輕鏈98.9%及重鏈89.5%。
基於人類生殖系基因使用以下V結構域對建構
λ - DP47文庫:Vl3_19 λ輕鏈與VH3_23重鏈。將輕鏈之CDR3 (L3)及重鏈之CDR3 (H3)中之文庫隨機化且藉由「重疊延長剪接」 (SOE) PCR自3個片段裝配。片段1包含包括隨機L3之抗體基因的5'端,片段2為自L3端跨越至H3的開始的中部恆定片段,而片段3包含Fab片段之隨機H3及3'部分。使用以下引子組合產生文庫之文庫片段:片段1 (LMB3-Vl_3_19_L3r_V/Vl_3_19_L3r_HV/Vl_3_19_L3r_HLV)、片段2 (RJH80-DP47CDR3_ba (mod))及片段3 (DP47-v4-4/DP47-v4-6/ DP47-v4-8-fdseqlong)。用於製造文庫片段之PCR參數為94℃下5分鐘初始變性,60秒94℃,60秒55℃,60秒72℃的25次循環及在72℃下10分鐘的末端延長。對於裝配PCR,使用等莫耳比的3個片段作為模板,參數為在94℃下3分鐘初始變性及60秒94℃、60秒55℃、120秒72℃的5次循環。在此階段,添加外部引子且進行額外20次循環,隨後72℃下的末端延長10分鐘。裝配足夠量的全長隨機Fab片段之後,將其用NcoI/NheI以及類似處理之受體噬菌粒載體消化。15 μg Fab文庫插入物用13.3 μg噬菌粒載體接合。經純化之接合用於60次轉型,產生1.5×109轉型體。呈現Fab文庫之噬菌粒粒子補救且藉由PEG/NaCl純化進行純化,以供選擇。
作為用於噬菌體呈現選擇之抗原的人類OX40 (CD134)在HEK EBNA細胞中短暫表現為N端單體Fc融合且在經位於攜帶受體鏈(Fc杵鏈)之Fc部分的C端的avi標籤識別序列處經BirA生物素接合酶之共表現活體內部位特異性生物素化。
根據以下模式,在溶液中進行數輪選擇(生物淘選):
1. 預清除經10 μg/ml不相關的人類IgG塗佈之maxisorp培養盤上約10
12個噬菌粒粒子以消耗識別抗原之Fc部分的抗體文庫,
2. 在100 nM不相關非生物素化Fc杵-臼構築體存在下用100 nM生物素化人類OX40培育非結合噬菌粒粒子0.5小時,進一步消耗總體積1 mL之Fc結合子,
3. 藉由轉移至經中性抗生物素蛋白預塗佈之微量滴定盤的4個孔中持續10分鐘,捕捉生物素化hu OX40及所連接之特異性結合噬菌體(在第1輪及第3輪中),
4. 使用5×PBS/Tween20及5×PBS洗滌相應的孔,
5. 藉由每孔添加250 μl 100 mM TEA (三乙胺)溶離噬菌粒粒子持續10分鐘,且藉由向來自4個孔之經合併之溶離液中添加500 μl 1 M Tris/HCl pH 7.4進行中和,
6. 藉由在經中性抗生物素蛋白預塗佈之微量滴定盤上與100 nM經生物素捕捉之Fc杵-臼構築體一起培育,後清除經中和之溶離液,以便最終移除Fc結合子,
7. 用經溶離之噬菌粒粒子的清液層再感染對數期大腸桿菌TG1細胞,用輔助噬菌體VCSM13感染,在震盪器上在30℃下培育隔夜且對下一輪選擇中所用之噬菌粒粒子進行後續PEG/NaCl沈澱。
使用100 nM之恆定抗原濃度進行3或4輪選擇。為了增加結合子不僅與獼猴OX40而且與鼠類OX40的交叉反應的可能性,在一些選擇回合中,使用鼠類目標代替人類OX40。在第2輪及第4輪中,為避免中性抗生物素蛋白之結合子增濃,捕捉抗原:藉由添加5.4×10
7抗生蛋白鏈菌素塗佈之磁珠進行噬菌體複合。藉由ELISA如下鑑別特異性結合子:100 μl 25 nM生物素化人類OX40及10 μg/ml人類IgG分別塗佈於中性抗生物素蛋白培養盤及maxisorp培養盤上。添加含Fab之細菌清液層且使用抗Flag/HRP二次抗體經由其Flag標籤偵測結合Fab。將在人類OX40上展現信號且在人類IgG上為陰性的純系列入候選清單進行進一步分析且亦以類似方式針對獼猴及鼠類OX40進行測試。其在0.5公升培養體積中經細菌表現,進行親和力純化且使用BioRad之ProteOn XPR36生物感測器藉由SPR分析進行進一步表徵。
表 3展示通用噬菌體呈現之抗體文庫(DP88-4)的序列,
表 4提供文庫DP88-4生殖系模板之cDNA及胺基酸序列且
表 5展示用於產生DP88-4生殖系模板的引子序列。
表 3 :通用噬菌體呈現抗體文庫(DP88-4)之序列
表 4 :文庫DP88-4生殖系模板之cDNA及胺基酸序列
表 5 :用於產生DP88-4文庫之引子序列
表 6展示通用噬菌體呈現之抗體共用輕鏈文庫(Vk3_20/VH3_23)的序列。
表 7提供共用輕鏈文庫(Vk3_20/VH3_23)生殖系模板之cDNA及胺基酸序列且
表 8展示用於產生共用輕鏈文庫(Vk3_20/VH3_23)之引子序列。
表 6 :用於PCR之通用噬菌體呈現抗體共用輕鏈文庫(Vk3_20/VH3_23)之序列
表 7 :共用輕鏈文庫(Vk3_20/VH3_23)生殖系模板之cDNA及胺基酸序列
表 8 :用於產生DP88-4文庫之引子序列
1:G/D = 20%,E/V/S = 10%,A/P/R/L/T/Y=5%;2:G/Y/S=15%,A/D/T/R/P/L/V/N/W/F/I/E = 4,6%;3:G/A/Y = 20%,P/W/S/D/T = 8%;4:F = 46%,L/M = 15%,G/I/Y = 8%;5:K=70%,R =30%。
表 9展示用於PCR之通用噬菌體呈現λ-DP47文庫(Vl3_19/VH3_23)模板之序列。
表 10提供λ-DP47文庫(Vl3_19/VH3_23)生殖系模板之cDNA及胺基酸序列且
表 11展示用於產生λ-DP47文庫(Vl3_19/VH3_23)之引子序列。
表 9 :用於PCR之通用噬菌體呈現λ-DP47文庫(Vl3_19/VH3_23)模板之序列
表 10 :λ-DP47文庫(Vl3_19/VH3_23)生殖系模板之cDNA及胺基酸序列
表 11 :用於產生λ-DP47文庫(Vl3_19/VH3_23)之引子序列
用於建構λ-DP47文庫之額外引子,即DP47CDR3_ba (mod.)、DP47-v4-4、DP47-v4-6、DP47-v4-8以及fdseqlong,與用於建構共用輕鏈文庫(Vk3_20/VH3_23)且已列於表8中之引子一致。
純系8H9、20B7、49B4、1G4、CLC-563、CLC-564及17A9經上文所述之程序鑑別為人類Ox40特異性結合子。其可變區之cDNA序列展示於下
表 12中,相應胺基酸序列可見於表C中。
表 12 :源自噬菌體之抗Ox40抗體之可變區鹼基對序列。帶下劃線的是互補決定區(CDR)。
1.3 抗 Ox40 IgG1 P329G LALA 抗體之製備、純化及表徵在具有人類IgG1之恆定重鏈或恆定輕鏈之框架中次選殖所選擇之抗Ox40結合子之重鏈及輕鏈DNA序列之可變區。根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入杵及臼重鏈之恆定區以消除與Fcγ受體之結合。
抗Ox40純系之cDNA及胺基酸序列展示於表13中。全部抗OX40-Fc融合編碼序列選殖至驅動來自MPSV啟動子之插入物的表現及含有位於CDS之3'端處的合成聚A信號序列的質體載體。此外,載體含有用於質體之游離型維護之EBV OriP序列。
表 13 : P329GLALA 人類 IgG1 型式中之抗 Ox40 純系之序列
藉由使用聚乙烯亞胺共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生所選擇之抗Ox40抗體。細胞用相應表現載體以1:1比率(「載體重鏈」:「載體輕鏈」)轉染。
對於在500 mL搖瓶中製造,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。轉染細胞在210×g下離心5分鐘,且清液層置換為預溫熱之CD CHO培養基。在20 mL CD CHO培養基中混合表現載體(200 μg完全DNA)。在添加540 μL PEI之後,溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。在轉染之後一天,添加1 mM丙戊酸及具有補充劑之7% Feed。培養7天之後,藉由在210×g下離心15分鐘來收集清液層。溶液無菌過濾(0.22 μm過濾器),用疊氮化鈉補充至最終濃度0.01%(w/v),且保持於4℃下。
使用用於抗原Fc融合物之純化的如上文所描述之蛋白質A,藉由親和層析進行抗體分子自細胞培養物清液層之純化。
濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl溶液(pH 6.0)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
使用基於胺基酸序列計算之莫耳消光係數,藉由在280 nm型量測OD來測定經純化之抗體之蛋白質濃度。使用LabChipGXII (Caliper)在存在及不存在還原劑(Invitrogen, USA)之情況下藉由CE-SDS分析抗體之純度及分子量。在25℃下,使用在25 mM K
2HPO
4、125 mM NaCl、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析抗體樣品之聚集物含量。
表 14概述抗Ox40 P329G LALA IgG1抗體之產量及最終含量。
表 14 :抗Ox40 P329G LALA IgG1純系之生物化學分析
實例 2 抗 OX40 抗體之表徵 2.1 人類 OX40 上之結合 2.1.1 表面電漿子共振 ( 親合力 + 親和力 )藉由表面電漿子共振(SPR)評定源自噬菌體之OX40特異性抗體與重組OX40 Fc(kih)之結合。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。
在同一實驗中,測定源自噬菌體呈現之抗OX40純系8H9、49B4、1G4、20B7、CLC-563、CLC-564及17A9 (全部人類IgG1 P329GLALA)以及重組OX40 (人類、獼猴及鼠類)之間的的物種選擇性及相互作用親合力。生物素化人類、獼猴及鼠類OX40 Fc (kih)直接偶合至抗生蛋白鏈菌素(SA)感測器晶片之不同流槽。使用至多600個共振單位(RU)之固定程度。
源自噬菌體呈現之抗OX40人類IgG1 P329GLALA抗體在2至500 nM範圍內之濃度(3倍稀釋)下以30 μL/min之流速,經120秒通過流槽。監測複合物解離210秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。
使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導動力學常數,以藉由數值積分針對1:1朗格繆爾結合擬合速率等式且用於定性評估親合力(
表 16)。
在同一實驗中,測定源自噬菌體呈現之抗體8H9、49B4、1G4、20B7、CLC-563及CLC-564 (人類IgG1 P329GLALA)與重組OX40之間的相互作用之親和力。為此目的,人類或鼠類Ox40之胞外域亦與avi (GLNDIFEAQKIEWHE)及六組胺酸標籤(序列參看
表 15)次選殖或藉由使用AcTEV蛋白酶裂解獲得且藉由層析法移除Fc。
表 15 :單體人類及鼠類Ox40 His標籤之核苷酸及胺基酸序列
如上文針對Fc融合蛋白所述進行蛋白質製造。分泌之蛋白質藉由螯合層析,隨後尺寸排阻層析自細胞培養物清液層純化。
在於20 mM磷酸鈉、500 nM氯化鈉,pH 7.4中平衡之NiNTA Superflow濾筒(5 ml,Qiagen)上進行第一層析步驟。藉由經12管柱體積施用5%至45%溶離緩衝液(20 mM磷酸鈉、500 nM氯化鈉、500 mM咪唑,pH 7.4)之梯度來進行溶離。
濃縮且過濾蛋白質,隨後裝載於用2 mM MOPS、150 mM氯化鈉、0.02% (w/v)疊氮化鈉溶液(pH 7.4)平衡之HiLoad Superdex 75管柱(GE Healthcare)上。
使用兩種設定進行親和力測定。
設定 1 )抗人類Fab抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約9000 RU。在25至50 nM濃度下捕捉針對OX40的源自噬菌體呈現之抗體持續60秒。重組人類OX40 Fc (kih)以4至1000 nM範圍內之濃度,以30 μL/min之流速,經120秒通過流槽。監測解離120秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fab抗體但已注射HBS-EP而非抗體之表面上流動。
設定 2 )抗人類Fc抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約8000 RU。在20 nM濃度下捕捉針對OX40的源自噬菌體呈現之抗體持續60秒。重組人類OX40 avi His以2.3至600 nM範圍內之濃度,以30 μL/min之流速,經120秒通過流槽。監測解離120秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fab抗體但已注射HBS-EP而非抗體之表面上流動。
使用Biacore T200評估軟體(vAA,Biacore AB, Uppsala/Sweden)獲得動力學常數,以藉由數值積分針對1:1朗格繆爾結合來擬合速率等式。
純系49B4、1G4及CLC-564以低於純系8H9、20B7及CLC-563之親和力結合人類Ox40 Fc(kih)。
藉由與1:1朗格繆爾結合擬合來測定抗OX40 P329GLALA IgG1與人類OX40 Fc (kih)之間的相互作用之親和力常數。
表 16 :抗OX40抗體與重組人類OX40之結合
2.1.2 結合於人類 Ox40 表現細胞 : 原生及經活化之人類周邊單核血液白細胞 ( PBMC )自Zürich血液供給中心獲得白血球層。為了分離新鮮的周邊血液單核細胞(PBMC),白血球層用相同體積之DPBS (Gibco,Life Technologies,目錄號14190 326)稀釋。提供50 mL聚丙烯離心管(TPP,目錄號91050)及15 mL Histopaque 1077 (SIGMA Life Science,目錄號10771,聚蔗糖及泛影酸鈉,調節至1.077 g/mL之密度)且經稀釋之白血球層溶液在Histopaque 1077上分層。試管在400×g下,在室溫下且在低加速度及不破裂情況下離心30分鐘。隨後,自界面收集PBMC,用DPBS洗滌三次且再懸浮於由以下組成之T細胞培養基中:具有10%胎牛血清(FBS,Gibco,Life Technology,目錄號16000-044,批號941273,γ照射、無黴漿菌且在56℃下熱不活化35分鐘)、1% (v/v)GlutaMAX I (GIBCO,Life Technologies,目錄號35050 038)、1 mM丙酮酸鈉(SIGMA,目錄號S8636)、1% (v/v)MEM非必需胺基酸(SIGMA,目錄號M7145)及50 µM β-巰基乙醇(SIGMA,M3148)之RPMI 1640培養基(Gibco,Life Technology,目錄號42401-042)。
PBMC在分離之後直接使用(原生人類PBMC上之結合)或其經刺激以接收T細胞之細胞表面上之強人類Ox40表現(經活化之人類PBMC上之結合)。因此,原生PBMC在6孔組織培養盤中的具有200 U/mL Proleukin及2 μg/mL PHA-L之T細胞培養基中培養五天,接著在37℃及5% CO
2下,在預塗佈之6孔組織培養盤[2 μg/mL 抗人類CD3 (純系OKT3)及2 μg/mL抗人類CD28 (純系CD28.2)]上在具有200 U/mL Proleukin之T細胞培養基中培養1天。
對於Ox40之偵測,混合原生人類PBMC與經活化之人類PBMC。為了能夠區分原生PBMC與經活化之人類PBMC,在結合分析之前使用eFluor670細胞增殖染料(eBioscience,目錄號65-0840-85)標記休眠細胞。
關於標記,採集細胞,用預溫熱(37℃)之DPBS洗滌且在DPBS中調整至1×10
7個細胞/毫升之細胞密度。eFluor670細胞增殖染料(eBioscience,目錄號65-0840-85)添加至原生人類PBMC於DPBS中之懸浮液(最終濃度為2.5 mM且最終細胞密度為0.5×10
7個細胞/毫升)中。接著細胞在室溫下,在黑暗中培育10分鐘。為了停止標記反應,添加2 mL FBS且用T細胞培養基洗滌各孔三次。接著將1×10
5個eFluor670標記之原生人類PBMC與未標記之經活化之人類PBMC的1:1混合物添加至圓底懸浮單元96孔培養盤(greiner bio-one,cellstar,目錄號650185)之各孔中。
培養盤用400×g且在4℃下離心4分鐘,且拂去清液層。細胞用200 μL 4℃冷FACS緩衝液(具有2% FBS、5 mM EDTA pH 8 (Amresco,目錄號E177)及7.5 mM疊氮化鈉(Sigma-Aldrich S2002))洗滌一次。細胞在4℃下在50 µL/孔含有經滴定之抗Ox40抗體構築體的4℃冷FACS緩衝液中培育120分鐘。培養盤用200 µL/孔4℃ FACS緩衝液洗滌四次,移除未經結合之構築體。
細胞在4℃下,在暗處,在25 µL/孔含有螢光標記之抗人類CD4 (純系RPA-T4,小鼠IgG1 k,BioLegend,目錄號300532)、抗人類CD8 (純系RPa-T8,小鼠IgG1k,BioLegend,目錄號3010441)、抗人類CD45 (純系HI30,小鼠IgG1k,BioLegend,目錄號304028)及異硫氰酸螢光素(FITC)結合之親和純化抗人類IgG Fcγ片段特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch,目錄號109-096-098)的4℃冷FACS緩衝液中染色30分鐘。
用200 µL/孔4℃ FACS緩衝液洗滌兩次的培養盤最終再懸浮於80 µL/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec, 目錄號Sc-3598)的FACS-緩衝液中且同一天使用5雷射LSR-Fortessa (BD Bioscience with DIVA software)獲得。
如
圖 2A - 2D中所示,沒有對OX40特異之抗體構築體結合於不表現OX40之休眠人類CD4
+T細胞或CD8
+T細胞。相比之下,全部構築體均結合於表現OX40的經活化之CD8
+或CD4
+T細胞。與CD4
+T細胞之結合比與CD8
+T細胞之結合牢固得多。經活化之人類CD8+ T細胞僅表現在經活化之CD4
+T細胞上偵測之OX40含量之一部分。差異為供體以及時間依賴性。分析之抗OX40純系的結合強度有變化。EC
50值顯示於
表 17中。對於靶向二價及單價FAP之構築體的進一步評估,選擇具有高(8H9)及低(49B4/1G4)結合能力之純系。
表 17 : 與經活化之人類 CD4 T 細胞結合的 EC
50 值
2.2 鼠類 OX40 上之結合 2.2.1 表面電漿子共振 ( 親合力 + 親和力 )藉由如上文針對人類OX40 Fc (kih)所述的表面電漿子共振(參看實例2.1.1)評定源自噬菌體之OX40特異性抗體20B7與重組鼠類OX40 Fc (kih)之結合。使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導動力學常數,以藉由數值積分針對1:1朗格繆爾結合擬合速率等式且用於定性評估親合力(
表 18)。
對於親和力測定,由於Fc融合蛋白與參考流槽之非特異性相互作用,因此使用以AcTEV蛋白酶裂解之鼠類Ox40 His (參看實例2.1.2)或Ox40 Fc(kih)。抗人類Fc抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約8000 RU。在25 nM濃度下捕捉針對OX40的源自噬菌體呈現之抗體持續60秒。重組鼠類OX40 (遵照經銷商說明藉由AcTEV消化裂解)在4.1至1000 nM範圍內之濃度下,以30 μL/min流速,經120秒通過流槽。監測解離120秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fab抗體但已注射HBS-EP而非抗體之表面上流動。
使用Biacore T200評估軟體(vAA,Biacore AB, Uppsala/Sweden)獲得動力學常數,以藉由數值積分針對1:1朗格繆爾結合來擬合速率等式。顯示純系20B7結合鼠類OX40 (
表 18)。
抗OX40 P329GLALA IgG1分子與鼠類OX40之間的相互作用之親和力常數使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導,藉由數值積分針對1:1朗格繆爾結合擬合速率等式。
表 18 :抗Ox40抗體20B7與鼠類OX40之結合
2.2.2 與小鼠 OX40 表現細胞之結合 : 原生及經活化之小鼠脾細胞 ( 所選純系 )在3 mL PBS中收集小鼠脾臟且使用溫和MACS試管(Miltenyi Biotec目錄號130-096-334)及gentleMACS Octo解離劑(Miltenyi Biotec)產生單細胞懸浮液。隨後,脾細胞經30 μm Pre-Separation過濾器(Miltenyi Biotec目錄號130-041-407)過濾且在350×g及4℃下離心7分鐘。抽吸清液層且細胞再懸浮於具有10% (v/v) FBS、1% (v/v) GlutaMAX-I、1 mM丙酮酸鈉、1% (v/v)MEM非必需胺基酸、50 μM β-巰基乙醇及10%青黴素-鏈黴素(SIGMA,目錄號P4333)之RPMI 1640培養基中。10
6個細胞/毫升在塗有10 μg/mL抗小鼠CD3ε Armenian Hamster IgG (純系145-2C11,BioLegend,目錄號100331)及2 μg/mL抗小鼠CD28 Syrian Hamster IgG (純系37.51,BioLegend,目錄號102102)之6孔組織培養盤中培養3天。收集經活化之小鼠脾細胞,在DPBS中洗滌,計數且將0.1×10
6個細胞轉移至96孔U型底經非組織培養物處理之盤之各孔中。細胞用DPBS洗滌且在50 μL含有不同濃度之抗OX40人類IgG1 P329GLALA抗體(僅所選結合子) 的FACS緩衝液中染色。細胞在4℃下培育120分鐘。接著,在4℃下,細胞用FACS緩衝液洗滌兩次且在25 μL/孔含有抗小鼠CD8b大鼠IgG2bκ-APC-Cy7 (BioLegend,目錄號100714,純系53-6.7)、抗小鼠CD4大鼠IgG2bκ-PE-Cy7 (BioLegend,目錄號100422,純系GK1.5)及FITC結合之AffiniPure抗人類IgG Fcγ-片段-特異性山羊IgG F(ab')
2片段(Jackson ImmunoResearch,目錄號109-096-098)之FACS緩衝液中染色30分鐘。用200 µL/孔4℃ FACS緩衝液洗滌兩次的培養盤最終再懸浮於80 µL/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec, 目錄號Sc-3598)的FACS-緩衝液中且同一天使用5雷射LSR-Fortessa (BD Bioscience with DIVA software)獲得。
如
圖 3A - 3D中所示,僅純系20B7及良好表徵之小鼠特異性基準抗體OX86顯示結合於經活化之小鼠CD4
+及CD8
+T細胞。休眠小鼠脾細胞上未觀測到結合。
2.3 獼猴 OX40 上之結合為了測試所選抗OX40結合子與獼猴細胞之反應性,使用如針對具有次要修飾之人類細胞所述之密度梯度離心分離健康食蟹獼猴之PBMC與肝素化血液。使用以DPBS稀釋之淋巴細胞分離劑培養基(90% v/v, Axon Lab, 目錄號1114545),用密度梯度離心分離獼猴PBMC與肝素化新鮮血液。在520×g下,在室溫下,不中斷的離心30分鐘。在室溫下,在150×g下相鄰離心15分鐘來減少血小板計數,隨後在室溫下進行400×g的若干離心步驟持續10分鐘,用無菌DPBS洗滌PBMC。刺激PBMC以接收T細胞之細胞表面上的強Ox40表現(經活化之獼猴PBMC上的結合)。因此,在37℃及5% CO
2下,原生PBMC在預塗佈之12孔組織培養盤[10 μg/mL獼猴交叉反應性抗人類CD3 (純系SP34)]及2 μg/mL獼猴交叉反應性抗人類CD28 (純系CD28.2)]上,在具有200 U/mL Proleukin之T細胞培養基中培養72小時。
0.5 × 10
5經活化之獼猴PBMC接著添加至圓底懸浮單元96孔培養盤(greiner bio-one, cellstar, 目錄號650185)的各孔中。細胞用200 μL 4℃冷FACS緩衝液洗滌一次且在4℃下,在50 µL/孔含有經滴定之抗Ox40抗體構築體的4℃冷FACS中培育120分鐘。接著,培養盤用200 µL/孔4℃ FACS緩衝液洗滌四次。將細胞再懸浮於25 µL/孔含有螢光標記之獼猴交叉反應性抗人類CD4 (純系OKT-4,小鼠IgG1 k,BD,目錄號317428)、抗人類CD8 (純系HIT8a,小鼠IgG1k,BD,目錄號555369)及FITC結合之AffiniPure抗人類IgG Fcγ-片段-特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch,目錄號109-096-098)的4℃冷FACS緩衝液中,且在4℃下在暗處培育30分鐘。用200 µL/孔4℃ FACS緩衝液洗滌兩次的培養盤最終再懸浮於80 µL/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec, 目錄號Sc-3598)的FACS-緩衝液中且同一天使用5雷射LSR-Fortessa (BD Bioscience with DIVA software)獲得。
如
圖 4A 及 4B中所示,大部分構築體結合於經活化之CD4
+獼猴T細胞。與CD4
+T細胞之結合比與CD8
+T細胞之結合牢固得多。OX40之表現水準視動力學及刺激強度而定,且針對CD4
+獼猴T細胞而非CD8
+獼猴T細胞最佳化,使得CD8
+T細胞上僅誘發極少OX40表現。所分析之抗OX40純系的結合強度發生變化。EC
50值顯示於
表 19中。由於非典型曲線擬合,純系8H9、49B4、21H4不可計算EC
50值。
表 19 : 與經活化之獼猴 CD4 T 細胞之結合的 EC
50 值
2.3.1 表面電漿子共振 ( 親合力 + 親和力 )藉由表面電漿子共振(SPR)評定源自噬菌體之OX40特異性抗體(全部人類IgG1 P329GLALA)與重組獼猴OX40 Fc(kih)之結合。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。
生物素化獼猴OX40 Fc(kih)直接偶合至抗生蛋白鏈菌素(SA)感測器晶片之不同流槽。使用至多800個共振單位(RU)之固定程度。
源自噬菌體呈現之抗OX40人類IgG1 P329GLALA抗體在2至500 nM範圍內之濃度(3倍稀釋)下以30 μL/min之流速,經120秒通過流槽。監測複合物解離210秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。
使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導動力學常數,以藉由數值積分針對1:1朗格繆爾結合擬合速率等式且用於定性評估親合力(
表 20)。
在同一實驗中,測定源自噬菌體呈現之抗體(人類IgG1 P329GLALA)與重組獼猴OX40 Fc(kih)之間的相互作用之親和力。抗人類Fab抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約9000 RU。在25至50 nM濃度下捕捉針對OX40的源自噬菌體呈現之抗體持續60秒。重組獼猴OX40 Fc (kih)以4至1000 nM範圍內之濃度,以30 μL/min之流速,經120秒通過流槽。監測解離120秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fab抗體但已注射HBS-EP而非抗體之表面上流動。
使用Biacore T200評估軟體(vAA,Biacore AB, Uppsala/Sweden)獲得動力學常數,以藉由數值積分針對1:1朗格繆爾結合來擬合速率等式(
表 20)。
純系49B4、1G4及CLC-564以低於純系8H9、20B7及CLC-563之親和力結合獼猴Ox40 Fc(kih)。
抗OX40 P329GLALA IgG1與獼猴OX40 Fc(kih)之間的相互作用之親和力常數使用Biacore T100評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導,藉由數值積分針對1:1朗格繆爾結合擬合速率等式。
表 20 :抗OX40抗體與重組獼猴OX40 Fc(kih)之結合
2.3.2 獼猴 OX40 表現細胞 : 經活化之獼猴周邊單核血液白細胞 ( PBMC ) 上的結合 與 OX40 陰性腫瘤細胞之結合使用WM266-4細胞(ATCC CRL-1676)及U-87 MG (ATCC HTB-14)腫瘤細胞測試與OX40陰性腫瘤細胞之結合的缺乏。為了分離兩種腫瘤細胞,WM266-4細胞用PKH-26紅色螢光細胞連接子試劑盒(Sigma,目錄號PKH26GL)預標記。採集細胞且用RPMI 1640培養基洗滌三次。糰粒在室溫下,在暗處,以最終細胞密度1×10
7個細胞在新鮮製備之PKH26-紅色染色溶液(所提供之稀釋劑C中最終濃度[1 nM])中染色5分鐘。添加過量FBS來停止標記反應且細胞用補充有10 % (v/v) FBS、1 % (v/v) GlutaMAX-I之RPMI 1640培養基洗滌四次,移除過量染料。
將5 × 10
4PKH26標記之WM266-4細胞及未經標記之U-87 MG細胞於DPBS中之混合物添加至96孔培養盤的圓底懸浮單元中。培養盤用400×g且在4℃下離心4分鐘,且拂去清液層。細胞用200 µL DPBS洗滌一次且糰粒藉由短且平緩渦流再懸浮。在4℃下,將全部樣品再懸浮於50 µL/孔含有滴定濃度之抗Ox40人類IgG1 P329GLALA抗體構築體的4℃冷FACS緩衝液中持續120分鐘。培養盤用200 µL/孔4℃ FACS緩衝液洗滌四次。將細胞再懸浮於25 µL/孔含有FITC結合之AffiniPure抗人類IgG Fcγ片段特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch,目錄號109-096-098)之4℃冷FACS緩衝液中且在4℃下在暗處培育30分鐘。用200 µL/孔4℃ FACS緩衝液洗滌兩次的培養盤最終再懸浮於80 µL/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec, 目錄號Sc-3598)的FACS-緩衝液中且同一天使用5雷射LSR-Fortessa (BD Bioscience with DIVA software)獲得。
如
圖 5A 及 5B中所示,沒有對OX40特異之抗體構築體結合於OX40陰性的人類腫瘤細胞WM266-4及U-78 MG。
2.4 配位體阻斷特性使用人類OX40配位體(R&D systems)測定OX40特異性人類IgG1 P329GLALA抗體分子干擾OX40/OX40-配位體相互作用的能力。由於OX40與OX40配位體之間的相互作用的低親和力,製備具有C端Ha標籤的二聚人類OX40 Fc融合物(
圖 1B)。此二聚人類Ox40 Fc融合分子的核苷酸及胺基酸序列顯示於
表 21中。如實例1.1中針對單體OX40 Fc(kih)所述進行製造及純化。
表 21 :二聚人類OX40 Fc融合分子之cDNA及胺基酸序列(由2個Fc鏈構成)
人類OX40配位體(R&D systems)使用標準胺偶合試劑盒(Biacore, Freiburg/Germany)在約2500 RU,pH 5.0下直接偶合至CM5晶片的兩個流槽。重組人類Ox40 Fc以200 nM濃度以30 μL/min的流速經90秒通過第二流槽。忽略解離且源自噬菌體的抗Ox40人類IgG1P329LALA以500 nM濃度以30 μL/min流速經90秒通過兩個流槽。監測解離60秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗體流經具有固定人類OX40配位體之表面,但上面已注射HBS-EP代替重組人類OX40 Fc。
圖 1C展示實驗設計。
源自噬菌體的純系20B7結合於人類OX40與其OX40配位體的複合物(
表 22 ,圖 6B)。因此,此抗體不與結合於人類OX40之配位體競爭且因此稱為「非配位體阻斷」。相反地,純系8H9、1G4、49B4、CLC-563及CLC-564不結合於人類OX40與其配位體之複合物且因此稱為「配位體阻斷」。
表 22 :表面電漿子共振測定之抗OX40純系之配位體結合特性
實例 3 抗人類 OX40 結合純系之功能特性 3.1 表現人類 OX40 及報導基因 NF - κ B - 螢光素酶之 HeLa 細胞OX40與其配位體之促效結合經核因子κB (NFκB)之活化誘導下游信號傳導(A. D. Weinberg等人, J. Leukoc. Biol. 2004, 75(6), 962-972)。產生重組型報導子細胞株HeLa_hOx40_NFκB_Luc1以在其表面上表現人類Ox40。此外,其具有受NFκB敏感性強化子片段控制之含有螢光素酶基因之報導子質體。Ox40觸發可誘導在細胞核中易位之NFκB之劑量依賴性活化,其中NFκB在報導子質體之NFκB敏感性強化子上結合以增加螢光素酶蛋白質之表現。螢光素酶催化螢光素氧化產生會發光的氧化螢光素。此可藉由光度計定量。一個實驗的範疇為測試多種抗Ox40結合子(P329GLALA huIgG1型式)在HeLa_hOx40_NFκB_Luc1報導體細胞中誘發NFκB活化的能力。
在37℃下,使用細胞解離緩衝液(Invitrogen,目錄號13151-014)經10分鐘收集黏著性HeLa_hOx40_NFκB_Luc1細胞。細胞用DPBS洗滌一次且在包含MEM (Invitrogen,目錄號22561-021)、10% (v/v)熱不活化FBS、1 mM丙酮酸鈉及1% (v/v)非必需胺基酸之分析培養基中調整至2×10
5個之細胞密度。細胞以0.3*10
5個細胞/孔之密度在具有蓋子之無菌白色96孔平底組織培養盤(greiner bio-one,目錄號655083)中接種,且在培育箱(Hera Cell 150)中保持在37℃及5% CO
2下隔夜。
第二天,添加含有多種經滴定之抗OX40結合子(P329GLALA huIgG1型式)之分析培養基刺激HeLa_hOX40_NFκB_Luc1持續6小時。關於測試抗OX40抗體上之超交聯作用,以1:2比率(二次抗體比初級單次抗OX40 P329GLALA huIgG1多2倍)添加50微升/孔含有二次抗體抗人類IgG Fcγ片段特異性山羊IgG F(ab')
2片段(Jackson ImmunoResearch,109-006-098)之培養基。在培育之後,抽吸清液層且培養盤用DPBS洗滌兩次。根據製造商說明,使用螢光素酶1000分析系統及報導子溶解緩衝液(皆來自Promega,目錄號E4550及目錄號E3971)進行發光之定量。簡言之,細胞藉由添加30微升/孔1×溶解緩衝液在-20℃下溶解10分鐘。細胞在37℃下解凍20分鐘,隨後添加90微升/孔螢光素酶分析試劑。立即藉由SpectraMax M5/M5e微板讀取器(Molecular Devices, USA)使用500 ms積分時間定量發光,無任何過濾器收集所有波長。所發射之相關光單元(URL)藉由HeLa_hOx40_NFκB_Luc1細胞之基本螢光校正且使用Prism4 (GraphPad Software, USA)針對對數一次抗體濃度點漬。使用嵌入式S形劑量反應擬合曲線。
如
圖 7A 及 7B中所示,已藉由向報導體細胞株添加抗OX40 P329GLALA huIgG1抗體(左側)來誘導有限劑量依賴性NFκB活化。抗OX40抗體與抗人類IgG特異性二次抗體的超交聯以濃度依賴性方式大大增加NFκB介導之螢光素酶活化的誘導(右側)。活化之EC
50值概述於
表 23中。
表 23 :與抗Ox40結合子(huIgG1 P329GLALA型式)及二次抗人類IgG Fcγ特異性抗體共培育之HeLa_hOx40_NFκB_luc1報導體細胞株中NFκB活化之EC
50值
3.2 OX40 介導之次佳 TCR 之共刺激觸發預活化之人類 CD4 T 細胞OX40之接合提供在次佳T細胞受體(TCR)刺激之後促進T細胞之分裂及存活的協同共刺激信號(M. Croft等人, Immunol. Rev. 2009, 229(1), 173-191)。另外,若干細胞激素之產量及T細胞活化標記之表面表現增加(I. Gramaglia等人, J. Immunol. 1998, 161(12), 6510-6517;S. M. Jensen等人., Seminars in Oncology 2010, 37(5), 524-532)。
為了測試多種抗OX40結合子之促效特性,經預活化之Ox40陽性CD4 T細胞用次佳濃度之培養盤固定之抗CD3抗體在溶液中或固定於培養盤表面上之抗OX40抗體存在下刺激72小時。藉由流動式細胞量測術,經由監測總細胞計數及活細胞中之CFSE稀釋來分析對T細胞存活及增殖之作用。另外,細胞用針對T細胞活化及分化標記的經螢光標識之抗體(例如CD127、CD45RA、Tim-3、CD62L及OX40本身)共染色。
人類PBMC經菲科爾密度離心(ficoll density centrifugation)分離且如0下所述用PHA-L [2 μg/mL]及Proleukin [200 U/mL]刺激三天。細胞接著在37℃下,用細胞密度1×10
6個細胞/毫升的CFSE用最終濃度[50 nM]之CFDA-SE (Sigma-Aldrich,目錄號2188)標記10分鐘。隨後,細胞用含有FBS (10% v/v)之過量DPBS洗滌兩次。經標記之細胞在T細胞培養基中在37℃下休眠30分鐘。隨後,用DPBS藉由兩個額外洗滌步驟移除未經轉化之CFDA-SE。根據製造商說明使用MACS陰性的CD4 T細胞分離試劑盒(Miltenyi Biotec)分離CD4 T細胞與預活化之CFSE標記之人類PBMC。
Morris等人顯示用習知抗Ox40抗體促效共刺激依賴於表面固定(N. P. Morris等人, Mol. Immunol. 2007, 44(12), 3112-3121)。因此,在4℃下,在有(表面固定之抗OX40)或無(溶液中之抗OX40)山羊抗人類Fcγ特異性抗體(Jackson ImmunoResearch,目錄號109-006-098)存在下,將山羊抗小鼠Fcγ特異性抗體(Jackson ImmunoResearch,目錄號111-500-5008)以PBS中[2 μg/mL]之濃度塗佈於96孔U型底細胞培養盤(Greiner Bio One)之表面上隔夜。此後,培養盤表面用含有BSA (1 % v/w)之DPBS阻斷。培育步驟之後全部T細胞在37℃下在含有BSA(1 % v/w)之PBS中處理90分鐘。在培育步驟之間,培養盤用DPBS洗滌。
小鼠抗人類CD3抗體(純系OKT3,eBioscience,目錄號16-0037-85,固定濃度[3 ng/mL])在隨後培育步驟中經表面塗佈之抗小鼠Fcγ特異性抗體捕捉。在一個實驗中,接著藉由DPBS中的額外培育步驟將經滴定之人類抗OX40抗體(人類IgG1 P329G LALA)固定於培養盤上。在第二實驗中,在活化分析期間將抗OX40抗體直接添加至培養盤中之培養基中,該培養盤未經抗人類IgG Fc特異性抗體預塗佈。
將經CFSE標記之預活化CD4
+T細胞以每孔0.6*10
5個細胞之細胞密度添加至預塗佈之培養盤中的200 μL T細胞培養基中且培養96小時。在4℃下,在暗處,細胞用螢光染料標記之小鼠抗人類Ox40 (純系BerACT35,Bioledgend,目錄號35008)、TIM-3 (純系F38-2E2,Biolegend,目錄號345008)、CD127 (純系A019D5,Biolegend,目錄號351234)、CD62L (純系DREG 56,Biolegend,目錄號304834)及CD45RA (純系HI100,BD Biosciences,目錄號555489)之組合染色20分鐘。用200 µL/孔4℃ FACS緩衝液洗滌兩次的培養盤最終再懸浮於80 µL/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec, 目錄號Sc-3598)的FACS-緩衝液中且同一天使用5雷射LSR-Fortessa (BD Bioscience with DIVA software)獲得。
分析DAPI陰性活細胞中作為增殖標記物之中位CFSE螢光的降低。監測作為T細胞活化標記物的OX40陽性、CD62L低及TIM-3陽性T細胞之百分比。分析CD45RA及CD127之表現來測定T細胞之成熟狀態的改變,由此將CD45RA低CD127低細胞分類為效應T細胞。
用培養盤固定之抗體共刺激以劑量依賴性方式大大提高預活化之人類CD4 T細胞用培養盤固定抗人類CD3之次佳刺激(
圖 8A - 8F)。T細胞增殖較強,顯示具有較高百分比之效應T細胞的更成熟表型且具有較高百分比之CD62L低、Tim-3陽性及OX40陽性活化細胞。一些純系(8H9、20B7)在結合於細胞OX40方面超過市售偵測抗體。沒有EC
50值計算對於其可行且因此OX40誘導之全部EC
50值均自整體EC
50值計算排除。T細胞活化之全部其他參數的半最大改變在3至700 pM範圍內之濃度下實現且概述於
圖 9及
表 24中。當在無表面固定存在下添加溶液中之抗Ox40抗體時,次佳TCR刺激未增強(
圖 10A - 10F)。此再次證實Ox40軸線活化對OX40受體之超交聯的高度依賴性。
抗OX40抗體(hu IgG1 P329GLALA型式)的結合強度與促效活性(生物活性)之間的相關性顯示於
圖 11中。對於多數純系而言,存在直接相關性,然而出乎意料地,兩個純系(49B4、1G4)顯示的生物活性比自其結合強度預測的生物活性強得多。
表 24 :用培養盤固定之抗OX40結合子(huIgG1 P329GLALA型式)補救次佳TCR刺激的EC
50值
實例 4 產生靶向 Ox40 及纖維母細胞活化蛋白 ( FAP ) 之雙特異性構築體 4.1 產生靶向 Ox40 及纖維母細胞活化蛋白 ( FAP ) 之雙特異性二價抗原結合分子 ( 2 + 2 型式 )製備對Ox40及FAP具有二價結合之雙特異性促效Ox40抗體。應用根據國際專利申請案第WO 2010/145792 A1號之互換單抗技術減少錯誤配對輕鏈之形成。
FAP結合子之產生及製備描述於WO 2012/020006 A2中,其以引用之方式併入本文中。
在此實例中,FAP結合子28H1之交叉Fab單元(VHCL)使用(G4S)
4連接體序列C端融合至抗OX40 huIgG1之重鏈。此重鏈融合與抗OX40之輕鏈及相應FAP交叉輕鏈(VLCH1)共表現。根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入重鏈之恆定區以消除與Fcγ受體之結合。所得雙特異性二價構築體描繪於
圖 12A中。
表 25分別展示成熟雙特異性二價抗OX40/抗FAP人類IgG1 P329GLALA抗體之核苷酸及胺基酸序列。
表 25 :雙特異性二價抗OX40/抗FAP人類IgG1 P329GLALA抗原結合分子之序列
全部基因在嵌合MPSV啟動子控制下短暫表現,該啟動子由MPSV核啟動子與CMV啟動子強化子片段之組合組成。表現載體亦含有針對含有EBNA(埃-巴二氏病毒核抗原)之宿主細胞中游離型複製之oriP區。
藉由使用聚乙烯亞胺共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生雙特異性抗Ox40抗FAP構築體。細胞用相應表現載體以1:1:1比率(「載體重鏈」:「載體輕鏈1」:「載體輕鏈2」)轉染。
關於在500 ml搖瓶中製造,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。關於轉染,使細胞在210×g下離心5分鐘,且用預溫熱之CD CHO培養基置換清液層。表現載體於20 ml CD CHO培養基中混合至200 μg DNA之最終量。在添加540 μL PEI之後,溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。轉染一天後,添加1 mM丙戊酸及7%饋料。在培養7天之後,藉由在210×g下離心15分鐘來收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
使用如上文針對抗原Fc融合物及抗體之純化所述的蛋白質A,藉由親和層析進行雙特異性構築體自細胞培養物清液層之純化。
濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl溶液(pH 6.0)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
藉由使用基於胺基酸序列計算之莫耳消光係數量測280 nm下的OD來測定經純化之雙特異性構築體的蛋白質濃度。使用LabChipGXII (Caliper)在有及無還原劑(Invitrogen, USA)存在下藉由CE-SDS分析雙特異性構築體之純度及分子量。在25℃下,使用在25 mM K
2HPO
4、125 mM NaCl、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析雙特異性構築體之聚集物含量(表26)。
表 26 :例示性雙特異性二價抗Ox40/抗FAP IgG1 P329G LALA抗原結合分子之生物化學分析
4.2 產生靶向 Ox40 及纖維母細胞活化蛋白 ( FAP ) 之雙特異性單價抗原結合分子 ( 1 + 1 型式 )藉由根據國際專利申請案第WO 2010/145792 A1號應用互換單抗技術製備對Ox40及FAP單價結合之雙特異性促效Ox40抗體來減少錯誤配對輕鏈之形成。
在此實例中,FAP結合子28H1之交叉Fab單元(VHCL)稠合至huIgG1之臼重鏈。針對抗Ox40之Fab融合至杵重鏈。靶向之抗FAP-Fc臼與抗Ox40-Fc杵鏈之組合允許產生雜二聚體,其包括FAP結合Fab及Ox40結合Fab (
圖 12B)。
根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入重鏈之恆定區以消除與Fcγ受體之結合。
所得雙特異性單價構築體描繪於
圖 12B中且核苷酸及胺基酸序列可見於
表 27中。
表 27 :成熟雙特異性單價抗Ox40/抗FAP huIgG1 P329GLALA kih抗體之cDNA及胺基酸序列
全部基因在嵌合MPSV啟動子控制下短暫表現,該啟動子由MPSV核啟動子與CMV啟動子強化子片段之組合組成。表現載體亦含有針對含有EBNA(埃-巴二氏病毒核抗原)之宿主細胞中游離型複製之oriP區。
藉由使用聚乙烯亞胺共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生雙特異性抗Ox40抗FAP構築體。細胞用相應表現載體以1:1:1:1比率(「載體重鏈臼」:「載體重鏈杵」:「載體輕鏈1」:「載體輕鏈2」)轉染。
關於在500 ml搖瓶中製造,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。關於轉染,使細胞在210×g下離心5分鐘,且用預溫熱之CD CHO培養基置換清液層。表現載體於20 ml CD CHO培養基中混合至200 μg DNA之最終量。在添加540 μL PEI之後,溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。轉染一天後,添加1 mM丙戊酸及7%饋料。在培養7天之後,藉由在210×g下離心15分鐘來收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
如上文針對抗原-Fc融合物及抗體之純化所述藉由親和層析使用蛋白質A自細胞培養物清液層純化雙特異性抗原結合分子。
濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl溶液(pH 6.0)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
藉由使用基於胺基酸序列計算之莫耳消光係數量測280 nm下的OD來測定經純化之雙特異性構築體的蛋白質濃度。使用LabChipGXII (Caliper)在有及無還原劑(Invitrogen, USA)存在下藉由CE-SDS分析雙特異性構築體之純度及分子量。在25℃下,使用在25 mM K
2HPO
4、125 mM NaCl、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析雙特異性構築體之聚集物含量。
表 28概述雙特異性單價抗Ox40/抗FAP IgG1 P329G LALA kih抗原結合分子之生物化學分析。
表 28 : 雙特異性單價抗 Ox40 / 抗 FAP IgG1 P329G LALA kih 抗原結合分子之生物化學分析
4.3 靶向 Ox40 及 FAP 之雙特異性構築體的表徵 4.3.1 表面電漿子共振 ( 同時結合 )藉由表面電漿子共振(SPR)評定同時結合人類Ox40 Fc (kih)及人類FAP之能力。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。生物素化人類Ox40 Fc (kih)與抗生蛋白鏈菌素(SA)感測器晶片之流動細胞直接偶合。使用至多1000個共振單位(RU)之固定程度。
靶向Ox40及FAP之雙特異性構築體在250 nM範圍內之濃度下以30 μL/min之流速經90秒穿過流槽且解離設為零秒。以250 nM之濃度,以30微升/分鐘之流速,經流槽經90秒注射人類FAP作為第二分析物(
圖 12C)。監測解離120秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。
如在
圖 13A - 13H之曲線中可見,所有雙特異性構築體可同時結合人類Ox40及人類FAP。
4.3.2 細胞上之結合 4.3.2.1 FAP 靶向抗 Ox40 抗體的與原生對比經活化之人類 PBMC 的結合如實例2.1.2中所述藉由ficoll密度離心來分離人類PBMC。PBMC在分離之後直接使用(休眠人類PBMC上之結合)或其經刺激以接收T細胞之細胞表面上之強人類Ox40表現(經活化之人類PBMC上之結合)。因此,在37℃及5% CO
2下,原生PBMC在6孔組織培養盤中在具有200 U/mL Proleukin及2 μg/mL PHA-L之T細胞培養基中培養四至七天,接著在經預塗佈之6孔組織培養盤[10 μg/mL抗人類CD3 (純系OKT3)及2 μg/mL抗人類CD28 (純系CD28.2)]中在具有200 U/mL Proleukin之T細胞培養基中培養1天。
對於Ox40之偵測,混合原生人類PBMC與經活化之人類PBMC。為了能夠區分原生PBMC與經活化之人類PBMC,在結合分析之前使用eFluor670細胞增殖染料(eBioscience,目錄號65-0840-85)標記原生細胞。接著將1×10
5個eFluor670標記之原生人類PBMC與未標記之經活化之人類PBMC的1:1混合物添加至圓底懸浮細胞96孔培養盤(greiner bio-one,cellstar,目錄號650185)之各孔中且如實例2.1.2中所述進行結合分析。接著將1×10
5個eFluor670標記之原生人類PBMC與未標記之經活化之人類PBMC的1:1混合物添加至圓底懸浮細胞96孔培養盤(greiner bio-one,cellstar,目錄號650185)之各孔中且如章節2.1.2中所述進行結合分析。
一次抗體為經滴定之抗Ox40抗體構築體,在4℃下培育120分鐘。二次抗體溶液為螢光標記之抗人類CD4 (純系RPA-T4,小鼠IgG1 k,BioLegend,目錄號300532)、抗人類CD8 (純系RPa-T8,小鼠IgG1k,BioLegend,目錄號3010441)及與異硫氰酸螢光素(FITC)結合之親和純化抗人類IgG Fcγ-片段特異性山羊IgG F(ab`)
2片段(Fluorescein isothiocyanate (FITC)-conjugated AffiniPure anti-human IgG Fcγ-fragment-specific goat IgG F(ab`)
2fragment )(Jackson ImmunoResearch,目錄號109-096-098)的混合物,其在4℃下在暗處培育30分鐘。最終,培養盤再懸浮於80微升/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec,目錄號Sc-3598)之FACS緩衝液中且在同一天使用5-雷射LSR-Fortessa (BD Bioscience with DIVA software)讀取。
如
圖 14A - 14P中所示,對Ox40特異之抗原結合分子不會與休眠人類CD4
+T細胞或CD8
+T細胞結合。反之,全部抗原結合分子均與經活化之CD8
+或CD4
+T細胞結合。與實例2.1.2中所述類似,與CD4
+T細胞之結合遠比與CD8
+T細胞之結合強力得多。如
圖 14A - 14P中所示,如習知IgG抗體型式之各別純系中,靶向二價FAP之Ox40構築體與Ox40陽性及陰性細胞顯示類似的結合特徵,而單價抗體與Ox40陽性細胞之結合能力由於親合力損失而明顯降低。
4.3.2.2 與表現人類 FAP 之腫瘤細胞之結合使用人類纖維母細胞活化蛋白(huFAP)表現細胞NIH/3T3-huFAP純系39或WM266-4細胞(ATCC CRL-1676)測試與細胞表面FAP之結合。藉由使用表現載體pETR4921轉染小鼠胚胎纖維母細胞NIH/3T3細胞株(ATCC CRL-1658)來表現huFAP,在1.5 μg/mL嘌呤黴素選拔下,產生NIH/3T3-huFAP純系39。在一些分析中,WM266-4細胞如實例2.3.2中所述,使用PKH-26紅色螢光細胞連接子試劑盒(Sigma,目錄號PKH26GL)預先標記,以便此等腫瘤細胞可與存在的其他細胞(例如人類PBMC)分離。
接著添加0.5 × 10
5個NIH/3T3-huFAP純系39或WM266-4細胞至圓底懸浮細胞96孔培養盤(greiner bio-one, cellstar,目錄號650185)的各孔中,且以與實例2.3.2中所述類似之方式進行結合分析。培養盤用400×g且在4℃下離心4分鐘,且拂去清液層。細胞用200 µL DPBS洗滌一次且糰粒藉由短且平緩渦流再懸浮。全部樣品再懸浮於50 µL/孔含有指定濃度範圍(經滴定)之雙特異性抗原結合分子(一次抗體)的4℃冷FACS緩衝液中且在4℃下培育120分鐘。隨後,細胞用200 µL 4℃ FACS緩衝液洗滌四次且藉由短渦流再懸浮。細胞用25 µL/孔含有異硫氰酸螢光素(FITC)結合之親和純化抗人類IgG Fcγ-片段-特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch, 目錄號109-096-098)的4℃冷二次抗體溶液進一步染色且在4℃下在暗處培育30分鐘。最終,培養盤再懸浮於80微升/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec,目錄號Sc-3598)之FACS緩衝液中且在同一天使用5-雷射LSR-Fortessa (BD Bioscience with DIVA software)獲取。
如
圖 15A 及 15B中所示,FAP靶向單價及二價抗Ox40抗原結合分子(但並非huIgG1 P329GLALA型式之相同純系)有效結合於人類FAP表現目標細胞。因此,僅FAP靶向單價及二價抗Ox40抗原結合分子顯示直接腫瘤靶向特性。二價構築體(實心正方形)顯示相較於單價構築體更牢固的與FAP結合,此由二價型式相對於單價型式之親合力增益解釋。此在高FAP表現NIH/3T3-huFAP純系39細胞(圖15A)中比較低FAP表現WM266-4細胞(圖15B)中更顯著。WM266-4細胞上較低密度之表面FAP可能不提供始終允許抗OX40構築體二價結合的FAP分子之緊密貼近。與經活化之人類CD4 T細胞及FAP陽性腫瘤細胞結合的EC
50值概述於
表 29中。
表29:FAP靶向單價或二價型式的所選aOx40結合子(純系8H9,1G4)與細胞表面人類FAP及人類Ox40之結合的EC50值
4.4 靶向 OX40 及纖維母細胞活化蛋白 ( FAP ) 且對 OX40 二價且對 FAP 單價 ( 2 + 1 型式 ) 之雙特異性抗原結合分子的產生藉由應用允許裝配兩種不同重鏈之杵-臼技術來製備對Ox40二價結合且對FAP單價結合之雙特異性促效Ox40抗體。
在此實例中,第一重鏈(HC 1)包含抗OX40結合子49B4之一個Fab單元(VHCH1),隨後為經(G
4S)連接子融合至抗FAP結合子28H1或4B9之VH結構域的Fc杵鏈。構築體之第二重鏈(HC 2)包含抗OX40結合子49B4之一個Fab單元(VHCH1),隨後為經(G
4S)連接子融合至抗FAP結合子28H1或4B9之VH結構域的Fc臼鏈。
FAP結合子之產生及製備描述於WO 2012/020006 A2中,其以引用之方式併入本文中。
根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入重鏈之恆定區以消除與Fcγ受體之結合。重鏈融合蛋白與抗OX40結合子49B4 (CLVL)之輕鏈共表現。對結合於OX40二價之所得雙特異性構築體描繪於
圖 12D中且核苷酸及胺基酸序列可見於表30中。
另外,製備「未靶向之」2+1構築體,其中抗FAP結合子之VH及VL結構域置換為不結合於抗原的生殖系對照物(稱為DP47)。
表 30 :成熟雙特異性二價抗Ox40/單價抗FAP huIgG1 P329GLALA kih抗體(2+1型式)及未靶向之(DP47)2+1構築體之cDNA及胺基酸序列
全部基因在嵌合MPSV啟動子控制下短暫表現,該啟動子由MPSV核啟動子與CMV啟動子強化子片段之組合組成。表現載體亦含有針對含有EBNA(埃-巴二氏病毒核抗原)之宿主細胞中游離型複製之oriP區。
使用聚乙烯亞胺(PEI; Polysciences Inc.)藉由用哺乳動物表現載體共轉染HEK293-EBNA細胞來製造雙特異性抗Ox40/抗FAP 2+1構築體。細胞用相應表現載體以1:1:2比率(「載體HC1」:「載體HC2」:「載體LC」)轉染。
對於500 mL搖瓶中的200 mL產量,250,000,000個HEK293 EBNA細胞接種24小時,隨後在具有補充劑之Excell培養基(Sigma)中轉染。關於轉染,使細胞在210×g下離心5分鐘,且用預溫熱之CD CHO培養基(Gibco)置換清液層。表現載體於20 mL CD-CHO培養基中混合至200 μg DNA之最終量。在添加540 μL PEI (1 mg/mL) (Polysciences Inc.)之後,將溶液渦旋15秒且在室溫下培育10分鐘。隨後,細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在37℃下,在5% CO
2氛圍下且在165 rpm下振盪下,在培育箱中培育3小時。培育後,添加160 mL具有補充劑(1 mM丙戊酸、5g/l PepSoy、6 mM L-麩醯胺酸)之Excell培養基且培養細胞24小時。轉染24小時之後,以12%最終體積(24 mL)及3 g/L葡萄糖(來自500 g/L原料的1.2 mL)的胺基酸及葡萄糖饋料補充細胞。在培養7天之後,藉由在2000-3000×g下離心45分鐘來收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
使用MabSelectSure藉由親和力層析法自細胞培養物清液層純化雙特異性構築體。濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl、0.01%Tween-20溶液(pH 6.0)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
關於親和力層析,將清液層裝載至經30 mL 20 mM檸檬酸鈉、20 mM磷酸鈉(pH 7.5)平衡的ProtA MabSelect Sure管柱(CV = 5 mL,GE Healthcare)上。藉由用6-10管柱體積之含有20 mM磷酸鈉、20 mM檸檬酸鈉(pH 7.5)的緩衝液洗滌來去除未經結合之蛋白質。未經結合之蛋白質使用分級或線性pH梯度的20 CV (0至100%)之20 mM檸檬酸鈉、100mM氯化鈉、100 mM甘胺酸、0.01% (v/v) Tween-20,pH 3.0溶離。管柱接著用10管柱體積之含有20 mM檸檬酸鈉、100 mM氯化鈉、100 mM甘胺酸、0.01%(v/v) Tween-20,pH 3.0之溶液洗滌,隨後進行再平衡步驟。
藉由添加1/10 (v/v) 0.5 M磷酸鈉,pH 8.0來調整所收集之溶離份的pH。濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl (pH 6.0)、0.01% Tween-20平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
藉由使用基於胺基酸序列計算之莫耳消光係數量測280 nm下的OD來測定經純化之雙特異性構築體的蛋白質濃度。使用LabChipGXII (Caliper)在有及無還原劑(Invitrogen)存在下藉由CE-SDS分析雙特異性構築體之純度及分子量。在25℃下,使用在25 mM磷酸鉀、125 mM氯化鈉、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析雙特異性構築體之聚集物含量(表31)。
表 31 :例示性雙特異性四價抗Ox40/抗FAP IgG1 P329G LALA抗原結合分子(2+1構築體)之生物化學分析
4.5 靶向 Ox40 及 FAP 之雙特異性 2 + 1 構築體的表徵 4.5.1 與 FAP 之結合 ( 表面電漿子共振 )藉由表面電漿子共振(SPR)評定雙特異性構築體結合人類、鼠類及獼猴FAP之能力。所有SPR實驗均在Biacore T200 (Biacore)上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20 (Biacore)來進行。
藉由以流動速率10 μL/min注射500 nM huFAP持續60秒、以流動速率20 μL/min注射10 nM鼠類FAP持續20秒且以以流動速率20 μL/min注射10 nM cynoFAP持續20秒,將His標記之人類、鼠類或食蟹獼猴二聚FAP捕捉於用抗His抗體(Qiagen目錄號34660)固定的CM5晶片(GE Healthcare)上。使用至多18000個共振單位(RU)之抗His抗體固定程度。分析之設定展示於
圖 12E中。
捕捉步驟之後,雙特異性構築體以及對照分子立即以0.78-100 nM範圍內之濃度,以30 μL/min之流動速率穿過晶片表面持續280秒,且解離期為180秒。藉由減去在參考流槽(其中無固定FAP)中獲得之反應來校正整體折射率差異。使用朗格繆爾1:1曲線擬合測定親和力。對於二價結合,使用相同1:1擬合產生表觀KD值。
表 32 :例示性雙特異性抗Ox40/抗FAP抗原結合分子與重組人類FAP、鼠類FAP及獼猴FAP之結合
注意:全部K
D均取決於特定實驗條件。
4.5.2 與人類 OX40 之結合 - 二價 Ox40 結合之競爭結合為了確認全部兩種抗OX40 Fab結構域結合於huOX40之能力與IgG相當,應用基於細胞之FRET分析法(TagLite)。因此,用huOX40-SNAP融合物轉染且用FRET供體鋱(Cisbio)標記之10000個Hek293 EBNA細胞/孔與0.2 nM用FRET受體d2 (Cisbio)標記之(49B4) IgG混合。另外,添加0.004-750 nM範圍之(49B4) IgG或雙特異性構築體49B4/28H1 (2+1)的濃度稀釋液且在室溫下培育2-4小時。在620 nm下量測螢光供體(鋱)之螢光信號且在665 nm下量測螢光受體染料(M100 Pro, Tecan)之螢光信號。計算665/620*1000之比率,且減去參考(僅細胞) (
圖 13L)。對於EC
50測定,在Graph Pad Prism5中分析結果。觀測之EC
50值顯示於表33中。在此等實驗條件下,全部2+1構築體均顯示與二價IgG類似之EC
50。
表 33 :IgG與二價2+1 OX40抗原結合分子之競爭結合的EC
50值;t=2h
4.5.3 與 OX40 及 FAP 之同時結合藉由表面電漿子共振(SPR)評定同時結合人類Ox40 Fc (kih)及人類FAP之能力。所有SPR實驗均在Biacore T200 (Biacore)上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20 (Biacore)來進行。
生物素化人類OX40 Fc (kih)與抗生蛋白鏈菌素(SA)感測器晶片之流動細胞直接偶合。使用至多1000個共振單位(RU)之固定程度。
靶向OX40及FAP之雙特異性抗體以250 nM之濃度,以流動速率30 μL/min穿過晶片表面持續90秒,且解離設為零秒。以濃度250 nM,以流動速率30 μL/min注射人類FAP持續90秒作為第二分析物(參看
圖 12F)。監測解離120秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。
全部雙特異性構築體均可同時結合於人類OX40及人類FAP (
圖 13I - 13K)。
4.5.4 細胞上之結合 4.5.4.1 靶向 OX40 及 FAP 之雙特異性抗體與原生人類 PBMC 之結合對比與經活化人類 PBMC 之結合如實例2.1.2中所述藉由ficoll密度離心來分離人類PBMC。PBMC在分離之後直接使用(休眠人類PBMC上之結合)或其經刺激以接收T細胞之細胞表面上之強人類Ox40表現(經活化之人類PBMC上之結合)。因此,原生PBMC在6孔組織培養盤中的具有200 U/mL Proleukin及2 μg/mL PHA-L之T細胞培養基中培養四天,接著在37℃及5% CO
2下,在預塗佈之6孔組織培養盤[4 μg/mL 抗人類CD3 (純系OKT3)及2 μg/mL抗人類CD28 (純系CD28.2)]上在具有200 U/mL Proleukin之T細胞培養基中培養1天。
對於OX40之偵測,混合原生人類PBMC與經活化之人類PBMC。為了能夠區分原生PBMC與經活化之人類PBMC,在結合分析之前使用eFluor670細胞增殖染料(eBioscience,目錄號65-0840-85)標記原生細胞。接著將1×10
5個eFluor670標記之原生人類PBMC與未標記之經活化之人類PBMC的1:1混合物添加至圓底懸浮細胞96孔培養盤(greiner bio-one,cellstar,目錄號650185)之各孔中且如實例2.1.2中所述進行結合分析。接著將1×10
5個eFluor670標記之原生人類PBMC與未標記之經活化之人類PBMC的1:1混合物添加至圓底懸浮細胞96孔培養盤(greiner bio-one,cellstar,目錄號650185)之各孔中且如章節2.1.2中所述進行結合分析。
一次抗體為經滴定之抗Ox40抗體構築體,在4℃下培育120分鐘。二次抗體溶液為螢光標記之抗人類CD4 (純系RPA-T4,小鼠IgG1 k,BioLegend,目錄號300532)、抗人類CD8 (純系RPa-T8,小鼠IgG1k,BioLegend,目錄號3010441)及與異硫氰酸螢光素(FITC)結合之親和純化抗人類IgG Fcγ-片段特異性山羊IgG F(ab`)
2片段 (Jackson ImmunoResearch,目錄號109-096-098)的混合物,其在4℃下在暗處培育60分鐘。最終,培養盤再懸浮於90微升/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec,目錄號Sc-3598)之FACS緩衝液中且在同一天使用5-雷射LSR-Fortessa (BD Bioscience with DIVA software)獲取。
如
圖 14I 及 14L中所示,無對Ox40特異之抗原結合分子結合於休眠人類CD4
+T細胞或CD8
+T細胞。相比之下,全部抗原結合分子均結合於經活化之CD8
+或CD4
+T細胞。與實例4.3.2.1中所述類似,與CD4
+T細胞之結合比與CD8
+T細胞之結合牢固得多。如
圖 14J 及 14L中所示,由於親合力增益,靶向二價FAP之OX40構築體顯示比作為單價抗體型式之各別純系的OX40陽性細胞牢固的結合特徵。全部型式的2+1設計獨立於第二特異性之結合部分以類似強度結合於OX40陽性細胞(
圖 14N 及 14P)。
4.5.4.2 與表現人類 FAP 之腫瘤細胞之結合與細胞表面FAP之結合使用人類纖維母細胞活化蛋白(huFAP)表現WM266-4細胞(ATCC CRL-1676)測試。使用表現NucLight Red螢光蛋白質之A549 NucLight™ Red細胞(Essenbioscience,目錄號4491)測試與OX40陰性FAP陰性腫瘤細胞之結合之缺乏,該NucLight Red螢光蛋白質受限於細胞核以允許自未標記之人類FAP陽性WM266-4細胞分離。根據標準Essen方案,親本A549 (ATCC CCL-185)在8 µg/ml凝聚胺存在下,用Essen CellPlayer NucLight Red慢病毒(Essenbioscience,目錄號4476;EF1α、嘌呤黴素)在MOI為3 (TU/細胞)之情況下轉導。此導致≥70%轉導效率。
將5 × 10
4未標記之WM266-4細胞及未標記之A549 NucLight™ Red細胞於FACS緩衝液中之混合物添加至圓底懸浮細胞96孔培養盤(greiner bio-one, cellstar,目錄號650185)之各孔中且以與實例4.3.2.2中所述類似之方式進行結合分析。培養盤用400×g且在4℃下離心4分鐘,且拂去清液層。細胞用200 µL DPBS洗滌一次且糰粒藉由短且平緩渦流再懸浮。全部樣品再懸浮於50 µL/孔含有指定濃度範圍(經滴定)之雙特異性抗原結合分子(一次抗體)的4℃冷FACS緩衝液中且在4℃下培育120分鐘。隨後,細胞用200 µL 4℃ FACS緩衝液洗滌四次且藉由短渦流再懸浮。細胞用25 µL/孔含有異硫氰酸螢光素(FITC)結合之親和純化抗人類IgG Fcγ-片段-特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch, 目錄號109-096-098)的4℃冷二次抗體溶液進一步染色且在4℃下在暗處培育60分鐘。最終,培養盤再懸浮於90微升/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec,目錄號Sc-3598)之FACS緩衝液中且在同一天使用5-雷射LSR-Fortessa (BD Bioscience with DIVA software)獲取。
如
圖 15C 及 15E中所示,FAP靶向單價及二價抗OX40抗原結合分子有效結合於人類FAP表現目標細胞。因此,僅FAP靶向單價及二價抗OX40抗原結合分子顯示直接腫瘤靶向特性。單價構築體之高親和力FAP結合純系4B9顯示比相同單價型式之各別FAP結合純系28H1更牢固的結合於人類FAP (
圖 15E)。不具有FAP結合域之2+1構築體未偵測到FAP (空心圓,
圖 15F)。與經活化之人類CD4
+T細胞及FAP陽性腫瘤細胞結合的EC
50值概述於表34中。
表 34 :FAP靶向單價或二價型式的aOx40結合子(純系49B4)與細胞表面人類FAP及人類Ox40之結合的EC
50值
實例 5 雙特異性抗人類 OX40 結合分子之功能特性 5.1 表現人類 OX40 及報導基因 NF κ B - 螢光素酶之 HeLa 細胞如實例3.1中所示,藉由向HeLa_hOX40_NFκB_Luc1報導體細胞株添加抗OX40 P329GLALA huIgG1抗體來誘導有限劑量依賴性NFκB活化。抗OX40抗體與抗人類IgG特異性二次抗體的超交聯以濃度依賴性方式大大增加NFκB介導之螢光素酶活化的誘導。因此,吾人測試單獨單價及二價FAP靶向型式且構築體經二次抗體或FAP
+腫瘤細胞株超交聯之所選抗OX40結合子(8H9、1G4、49B9)的NFκB活化能力。
如實例3.1中所述,附著之HeLa_hOX40_NFκB_Luc1細胞在每孔0.3×10
5個細胞的細胞密度下培養隔夜且用含有經滴定之FAP靶向單價(FAP 1+1)及二價(FAP 2+2)型式且呈P329GLALA hu IgG1構築體形式的抗OX40結合子(純系8H9及1G4)的分析培養基刺激5至6小時。為了測試二次抗體之超交聯作用,以1:2比率(一次抗體比二次抗體)添加25 μL/孔含有二次抗體抗人類IgG Fcγ-片段-特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch,109-006-098)之培養基。為了測試藉由細胞表面FAP結合進行之超交聯之作用,25微升/孔含有FAP
+腫瘤細胞(WM266-4及/或NIH/3T3-huFAP純系39)之培養基以2:1比率(每個孔中FAP
+腫瘤細胞比報導子細胞多兩倍)共同培養。藉由如實例3.1中所述使用螢光素酶1000分析系統及報導體溶解緩衝液(皆Promega,目錄號E4550及目錄號:E3971)量測發光來定量經活化之NFκB。
如
圖 16A - 16G中所示,全部抗OX40構築體之存在均誘發有限NFκB活化。經二次抗huIgG Fcγ-特異性抗體超交聯提高huIgG1 P329GLALA以及FAP靶向單價/二價型式之結合子的此NFκB活化。與OX40之單價結合由此比與OX40之二價結合低效,顯示OX40受體寡聚對完全活化OX40信號傳導軸的必要性。當使用FAP靶向分子(實心正方形及三角形)時,FAP表現腫瘤細胞以濃度依賴性方式大大提高NFκB介導之螢光素酶活化的誘導。當使用非靶向huIgG1 P329GLALA型式之相同純系時未發現此類作用,因為構築體不能經FAP
+腫瘤細胞進一步超交聯。二價分子又優於單價分子。FAP之高度表現確保較高交聯且因此FAP靶向構築體更好的促效作用(比較FAP高NIH/3T3-huFAP純系39及FAP陽性WM266-4細胞的實心菱形)。二價FAP靶向之構築體顯示在FAP
+腫瘤細胞存在下約0.1至1 nM濃度下的峰值活性。化合物濃度之另一增加實際上降低其誘導NFκB之能力。最可能地,結合於僅一個目標(FAP或OX40)之二價構築體以較高濃度存在,勝出構築體同時結合於FAP及OX40。此交聯損失又減少促效OX40信號傳導。
在另一實驗中,附著之HeLa_hOX40_NFκB_Luc1細胞在每孔0.2×10
5個細胞的細胞密度下培養隔夜且用含有經滴定之FAP靶向單價(FAP 1+1)及二價(FAP 2+1及FAP 2+2)型式且呈P329GLALA hu IgG1構築體形式的抗OX40結合子(純系49B9)的分析培養基刺激5小時。2+1之FAP經兩種不同FAP結合純系靶向,28H1以低親和力靶向FAP且4B9以高親和力靶向FAP。為了測試二次抗體之超交聯作用,以1:2比率(一次抗體比二次抗體)添加25 μL/孔含有二次抗體抗人類IgG Fcγ-片段-特異性山羊IgG F(ab`)
2片段(Jackson ImmunoResearch,109-006-098)之培養基。為了測試藉由細胞表面FAP結合進行之超交聯之作用,25微升/孔含有FAP
+腫瘤細胞(NIH/3T3-huFAP純系19)之培養基以4:1比率(每個孔中FAP
+腫瘤細胞比報導子細胞多四倍)共同培養。藉由如實例3.1中所述使用螢光素酶1000分析系統及報導體溶解緩衝液(皆Promega,目錄號E4550及目錄號:E3971)量測發光來定量經活化之NFκB。
如
圖 16H - 16M中所示,在此實驗中全部抗OX40構築體之存在亦誘導有限NFκB活化。經二次抗huIgG Fcγ特異性抗體超交聯增加FAP靶向單價/二價型式之全部結合子的此NFκB活化。與OX40之單價結合由此比與OX40之二價結合低效,顯示OX40受體寡聚對完全活化OX40信號傳導軸的必要性(
圖 16I,比較EC
50值)。當使用FAP靶向分子(實心正方形、三角形、半實心圓形)時,FAP表現腫瘤細胞以濃度依賴性方式大大提高NFκB介導之螢光素酶活化的誘導。當2+1型式之FAP結合部分經非結合DP47單元(空心圓)置換時未發現此類作用,因為構築體不能經FAP
+腫瘤細胞進一步超交聯。二價分子又優於單價分子(比較EC
50值)。單價FAP靶向及二價OX40靶向構築體(2+1)在FAP
+腫瘤細胞存在下顯示最高平穩活性。與單價OX40結合1+1構築體相比,與2+1型式之OX40二價結合提高OX40寡聚之促效能力(對EC
50值的影響)。然而,由於2+1構築體與FAP之單價結合,看起來相較於二價FAP結合2+2型式可交聯兩倍的分子,此可解釋較高平穩OX40活化(
圖 16J,較高平穩階段)。
5.2 OX40 介導之次佳 TCR 之共刺激觸發預活化之人類 CD4 T 細胞亦測試FAP靶向單價或二價型式之所選結合子(純系8H9及1G4)共活化表面固定之T細胞的能力。
如實例3.2中所述,經預先活化之CFSE標記之OX40陽性CD4 T細胞在固定於培養盤表面的經滴定之抗Ox40抗體存在下用次佳濃度之培養盤固定之抗CD3抗體刺激72小時。藉由流動式細胞量測術,經由監測總細胞計數及活細胞中之CFSE稀釋來分析對T細胞存活及增殖之作用。另外,細胞用針對T細胞活化及分化標記的經螢光標識之抗體(例如CD127、CD45RA、Tim-3、CD62L及OX40本身)共染色。
用培養盤固定之雙特異性抗OX40抗原結合分子共刺激以劑量依賴性方式大大提高經預先活化之人類CD4 T細胞用培養盤固定之抗人類CD3的次佳刺激(
圖 17A 及 17B)。更有效增殖之T細胞顯示具有較高百分比之CD127低T細胞、Tim-3陽性及OX40陽性活化細胞的更成熟表型(純系8H9僅顯示粒度(SSC)增加)。與OX40之單價結合(三角形符號)由此比與OX40之二價結合(正方形符號)低效,顯示OX40受體寡聚對完全活化Ox40信號傳導軸的必要性。T細胞活化之全部分析參數中的半最大改變在3至2300 pM範圍內之濃度下實現且針對純系1G4概述於
圖 18A - 18D及
表 35中。
表 35 :用培養盤固定之FAP靶向單價及二價抗Ox40 (純系1G4)構築體補救次佳TCR刺激的EC
50值
5.3 OX40 介導之次佳 TCR 觸發之休眠人類 PBMC 及藉由細胞表面 FAP 進行之超交聯之共刺激實例5.1中顯示添加FAP
+腫瘤細胞可藉由提供OX40受體之強寡聚大大提高人類OX40陽性報導體細胞株中FAP靶向單價及二價抗OX40構築體誘導之NFκB活性。同樣,吾人在NIH/3T3-huFAP純系39細胞存在下測試FAP靶向單價(1+1)及二價(2+1,2+2)抗OX40構築體補救休眠人類PBMC細胞之次佳TCR刺激的能力。
人類PBMC製劑含有(1)休眠OX40陰性CD4
+及CD8
+T細胞及(2)在細胞表面上具有多種Fcγ受體之分子之抗原呈現細胞,例如B細胞及單核細胞。人類IgG1同型之抗人類CD3抗體可藉由其Fc部分結合於本發明之Fc-γ受體分子且介導休眠Ox40陰性CD4
+及CD8
+T細胞上之延長TCR活化。接著,此等細胞開始在若干小時內表現Ox40。針對OX40之功能性促效化合物可經由經活化之CD8
+及CD4
+T細胞上呈現之OX40受體進行信號傳導及支持TCR介導之刺激。
休眠的經CFSE標記之人類PBMC用次佳濃度之抗CD3抗體在輻射FAP
+NIH/3T3-huFAP純系39細胞及經滴定之FAP-Ox40構築體存在下刺激四至五天。藉由流動式細胞量測術,經由監測總細胞計數及活細胞中之CFSE稀釋來分析對T細胞存活及增殖之作用。另外,細胞用針對T細胞活化及成熟標記物CD25、顆粒酶B、CD62L及CD127之螢光標記之抗體共染色。在第二實驗中,細胞用針對T細胞活化及成熟標記物CD25及Tim-3之螢光標記之抗體共染色。
在37℃下,使用細胞解離緩衝液(Invitrogen,目錄號13151-014)收集小鼠胚胎纖維母細胞NIH/3T3-huFAP純系39細胞(參看實例4.3.2.2)持續10分鐘。細胞用DPBS洗滌一次。NIH/3T3-huFAP純系39細胞在培育箱(Hera Cell 150)中,在37℃及5% CO
2下,在無菌96孔圓底黏著組織培養盤(TPP,目錄號92097)中之T細胞培養基中以0.2×10
5個細胞/孔之密度培養隔夜。第二天,其在xRay輻射器中使用4500 RAD之劑量輻射以防止稍後由腫瘤細胞株引起之人類PBMC之過度生長。
人類PBMC藉由ficoll密度離心分離且如實例2.1.2中所述用CFSE標記。細胞以0.75×10
5個細胞/孔(第一實驗)或0.5×10
5個細胞/孔(第二實驗)之密度添加至各孔中。添加最終濃度[10 nM]之抗人類CD3抗體(純系V9,人類IgG1)及指定濃度的FAP靶向單價及二價抗OX40抗原結合分子。細胞在培育箱(Hera Cell 150)中在37℃及5% CO
2下活化四至五天。接著,細胞在4℃下用螢光染料結合之抗體抗人類CD4 (純系RPA-T4, BioLegend, 目錄號300532)、CD8 (純系RPa-T8, BioLegend, 目錄號3010441)、CD25 (純系M-A251, BioLegend, 目錄號356112)、CD127 (純系A019D5, Biolegend, 目錄號351234)及CD62L (純系DREG 56, Biolegend, 目錄號304834)或Tim-3 (純系F38-E2E, Biolegend, 目錄號345012)表面染色30分鐘。對於透化細胞膜,細胞糰粒用FACS緩衝液洗滌兩次,接著在室溫下在暗處再懸浮於50微升/孔新鮮製備之FoxP3 Fix/Perm緩衝液(eBioscience目錄號00-5123及00-5223)中45分鐘。用Perm-洗滌緩衝液(eBioscience, 目錄號00-8333-56)洗滌三次後,細胞在室溫下在暗處用25 μL/孔含有抗人類顆粒酶B抗體(純系GB-11, BD Bioscience, 目錄號561142)之Perm-洗滌緩衝液胞內染色1小時。細胞再懸浮於85微升/孔FACS緩衝液中且使用5-雷射Fortessa流式細胞儀(BD Bioscience with DIVA software)獲取。
如
圖 20A - 20H中所示,用非靶向抗Ox40 (8H9) huIgG1 P329GLALA (空心正方形)共刺激不會補救次佳TCR刺激之CD4及CD8 T細胞。FAP靶向單價(實心三角形)及二價(實心正方形)抗OX40構築體經本發明NIH/3T3-huFAP純系39細胞超交聯大大提高增殖、存活率且在人類CD4及CD8 T細胞中誘導強化之經活化表型。對於高親和力純系8H9 (
圖 20A - 20H),單價及二價雙特異性抗原結合分子具有補救次佳TCR刺激之相當能力。兩種構築體皆顯示在約0.1-1 nM下的峰值活性及在較高濃度下降低之反應。與NFκB報導體細胞株(
圖 16A - 16M)中之發現類似,此可為僅結合於FAP或OX40之構築體與同時結合於兩個目標的構築體之間的目標結合之競爭結果,且因此提供必要交聯。當低親和力純系1G4在FAP靶向單價對比二價抗OX40構築體中測試時,此作用較不顯著(
圖 21A - 21C)。此處,單價抗體明顯次於二價構築體。當各構築體之促效能力定量為曲線下面積且針對彼此繪製時(
圖 21A - 21C),此可最佳理解。
第二實驗中獲得之資料分別顯示於
圖 21D - 21H 及 21I - 21M中。如
圖 21D - 21H中所示,用非靶向抗Ox40 (49B4) 2+1 DP47型式(空心圓)共刺激不會補救次佳TCR刺激之CD4及CD8 T細胞。FAP靶向單價(實心三角形)及二價(實心正方形、半實心圓)抗OX40構築體經本發明NIH/3T3-huFAP純系39細胞超交聯大大提高存活率且在人類CD4及CD8 T細胞中誘導強化之經活化表型。單價抗OX40構築體(1+1;實心三角形)補救TCR刺激的能力低於二價抗OX40靶向構築體(半實心圓,實心正方形)。FAP與2+2構築體之二價結合相較於FAP與2+1構築體之單價結合已經能夠在較低濃度下補救次佳TCR刺激。在2+1型式中,高親和力FAP結合純系4B9明顯優於低親和力純系28H1 (
圖 21I - 21M)。此表明觀測到的生物活性的EC
50值由與FAP之結合(2+2 > 2+1 (4B9) > 2+1 (28H1))驅動。當各構築體之促效能力針對作為曲線下面積之分析標記定量且針對彼此繪製時(
圖 21N),此可最佳理解。
5.4 藉由使用人類 IgG1 P329GLALA 型式預防 ADCC人類IgG1類抗體可藉由結合於ADCC勝任細胞(例如NK細胞)上之Fcγ受體(FcγR)誘發抗原陽性目標細胞之抗體依賴性細胞死亡(ADCC)。因此,ADCC勝任Ox40抗體可介導最近活化之Ox40
+T細胞的溶解且減少腫瘤反應性T細胞之彙集。向抗體之Fc部分引入IgG1P329GLALA突變防止結合於FcγR (國際專利申請公開案第WO 2012/130831 A1號),且因此在NK細胞存在下ADCC。然而,與FcN受體之結合未經改變以確保抗體之IgG樣藥物動力學。
所選純系轉化成習知人類IgG1型式來測試其誘發ADCC之能力。為了測試ADCC競爭力,PKh26標記之OX40陽性腫瘤細胞(HeLa_hOX40_NFκB_Luc1報導體細胞株)以及新鮮分離之NK細胞在抗OX40抗體(人類IgG1或人類IgG1-P329GLALA型式)之連續稀釋列存在下在3比1之E比T比率下培養。使用乳酸脫氫酶(LDH)之釋放及DAPI陽性腫瘤細胞之量定量NK細胞介導之ADCC。
簡言之,HeLa_hOX40_NFκB_Luc1細胞(實例3.1)如實例2.3.2中所述使用PKH-26紅色螢光細胞連接子試劑盒(Sigma, 目錄號PKH26GL)標記。在37℃及5% CO
2下,在培育箱(Hera細胞150)中,PKH-26標記之HeLa_hOx40_NFκB_luc1細胞以0.5×10
5個細胞/孔之密度接種於無菌96孔圓底黏著組織培養盤(TPP, 目錄號92097)中之AIM V培養基(Gibco, 目錄號12055-09)中隔夜。第二天,人類PBMC如實例2.1.2中所述藉由ficoll密度梯度離心分離。根據製造商說明使用MACS陰性的NK細胞分離試劑盒,人類(Miltenyi Biotec,目錄號130-092-657)進行NK細胞分離。以AIM V培養基中1.5×10
5個細胞/孔之密度添加NK細胞,導致3比1之E比T比率。以指定濃度添加抗OX40抗體(人類IgG1或人類IgG1 P329GLALA)且培養盤在37℃及5% CO
2下在培育箱(Hera Cell 150)中培育隔夜。4小時後,取樣100 μL清液層用於LDH分析且培養基更換成新鮮AIM-V培養基。在SpectraMax M5/M5
e(Molecular Devices)微板讀取器(Filter 490 nm - 650 nM, 1ms積分時間)上根據製造商說明使用細胞毒性偵測試劑盒-LDH (Roche, 目錄號11644793001)定量LDH活性。
24小時後,使用胰蛋白酶剝落細胞。細胞用螢光標記之抗人類CD56 (純系NCAM16.2,小鼠IgG1 κ, BioLegend, 目錄號562751)、抗人類CD25 (純系MA251,小鼠IgG1 κ, BioLegend, 目錄號356116)及抗人類CD69 (純系FN50, 小鼠IgG1 κ, BioLegend, 目錄號356116)染色,在4℃在暗處培育20分鐘。最終,培養盤再懸浮於80微升/孔含有0.2 μg/mL DAPI (Santa Cruz Biotec,目錄號Sc-3598)之FACS緩衝液中且在同一天使用5-雷射LSR-Fortessa (BD Bioscience with DIVA software)獲取。
ADCC勝任人類IgG1型式之全部Ox40抗體能夠誘發與針對EGFR之ADCC勝任參考抗體(GA201)類似程度之Ox40陽性細胞溶解。IgG1 P329GLALA型式之純系不介導ADCC (參看
圖 22A 及 22B)。
向吾人靶向型式之Fc部分引入IgG1 P329GLALA突變防止與FcγR之結合(國際專利申請公開案第WO 2012/130831 A1號),且因此在NK細胞存在下ADCC。然而,與FcN受體之結合未經改變以確保抗體之IgG樣藥物動力學。與已存在之OX40抗體相比,藉由結合於FAP陽性腫瘤細胞或纖維母細胞在本發明之雙特異性抗體中提供最佳促效OX40信號傳導所必需之超交聯。許多腫瘤適應症均報導腫瘤相關纖維母細胞或腫瘤細胞本身上之FAP陽性。因此,雙特異性型式在腫瘤微環境中在無全身性活化存在下具有強OX40介導之促效作用的可能,此可預防免疫相關毒性。與習知抗OX40抗體相反,本發明之雙特異性抗原結合分子不誘發最近活化之OX40陽性效應T細胞的ADCC。因此,雙特異性抗體可具有再活化先前存在但經抑制的針對腫瘤細胞之適應性免疫反應的潛力且可有效用於治療癌症患者。
實例 6 產生 4 - 1BB 抗體及工具結合子 6.1 抗原之製備、純化及表徵以及藉由噬菌體呈現產生新 4 - 1BB 結合子的篩選工具編碼人類、小鼠或獼猴4-1BB之胞外域的DNA序列(
表 36)與人類IgG1重鏈CH2及CH3結構域在杵上同框次選殖(Merchant等人, 1998)。在抗原胞外域與人類IgG1之Fc之間引入AcTEV蛋白酶裂解位點。在抗原-Fc杵之C端引入用於引導生物素化之Avi標籤。含有S354C/T366W突變之抗原-Fc杵鏈與含有Y349C/T366S/L368A/Y407V突變之Fc臼鏈之組合允許產生雜二聚體,其包括含有4-1BB胞外域之鏈之單一複本,因此產生Fc連接之抗原之單體形式(
圖 1A)。
表 37展示抗原Fc-融合物構築體之cDNA及胺基酸序列。
表 36 :抗原胞外域(ECD)之胺基酸編號及其來源
表 37 :單體抗原Fc(kih)融合分子之cDNA及胺基酸序列(藉由組合一個Fc臼鏈與一個抗原Fc杵鏈製備)
將所有4-1BB-Fc-融合物分子編碼序列選殖入質體載體,其驅動插入物自MPSV啟動子之表現且含有位於CDS之3'端處的合成polyA信號序列。此外,載體含有用於質體之游離型維護之EBV OriP序列。
關於生物素化單體抗原/Fc融合分子之製備,用三個編碼融合蛋白之兩個組分(結及臼鏈)的載體以及BirA (生物素化反應所需之酶)共轉染以指數方式生長之懸浮液HEK293 EBNA細胞。以2:1:0.05比率(「抗原ECD-AcTEV-Fc杵」:「Fc臼」:「BirA」)使用相應載體。
關於在500 ml搖瓶中製造蛋白質,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。關於轉染,使細胞在210 g下離心5分鐘,且用預溫熱之CD CHO培養基置換清液層。使表現載體再懸浮於20 mL含有200 μg載體DNA之CD CHO培養基中。在添加540 μL聚乙烯亞胺(PEI)之後,將溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。製備培養基補充有5 μM基夫鹼(kifunensine)。在轉染之後一天,向培養物中添加1 mM丙戊酸及7%饋料。培養7天之後,藉由使細胞在210 g下短暫離心15分鐘收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
藉由使用蛋白質A進行親和層析,接著進行尺寸排阻層析自細胞培養物清液層純化所分泌之蛋白質。關於親和力層析,將清液層裝載至經40 mL 20 mM磷酸鈉、20 mM檸檬酸鈉(pH 7.5)平衡的HiTrap蛋白A HP管柱(CV = 5 mL,GE Healthcare)上。藉由用至少10管柱體積之含有20 mM磷酸鈉、20 mM檸檬酸鈉及0.5 M氯化鈉之緩衝液(pH 7.5)洗滌來移除未結合之蛋白質。使用經20管柱體積之20 mM檸檬酸鈉、0.01% (v/v)Tween-20 (pH 3.0)產生之氯化鈉之線性pH梯度(0至500 mM)來溶離結合之蛋白質。接著用10管柱體積之20 mM檸檬酸鈉、500 mM氯化鈉、0.01% (v/v) Tween-20 (pH 3.0)洗滌管柱。藉由添加1/40 (v/v) 2 M Tris,pH 8.0來調整所收集之溶離份之pH。濃縮且過濾蛋白質,隨後裝載於用2 mM MOPS、150 mM氯化鈉、0.02% (w/v)疊氮化鈉溶液(pH 7.4)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
6.2 自通用 F ( ab ) 庫選擇 4 - 1BB 特異性 12B3 、 25G7 、 11D5 、 9B11 及 20G2 抗體對人類及獼猴4-1BB具有特異性之抗體
11D5、
9B11及
12B3選自Fab型式之通用噬菌體呈現抗體文庫(DP88-4)。亦自同一文庫選擇對鼠類4-1BB具有反應性的額外抗體純系
20G2。此文庫基於人類生殖系基因使用V結構域成對Vk1_5 (κ輕鏈)及VH1_69 (重鏈)建構,其包含輕鏈之CDR3 (L3,3種不同長度)及重鏈之CDR3 (H3,3種不同長度)中的隨機序列間隔。藉由重疊延長(SOE) PCR施加剪接,藉由3 PCR擴增之片段的組合件進行文庫繼代。片段1包含包括隨機L3之抗體基因的5'端,片段2為自L3跨越至H3的中部恆定片段而片段3包含抗體基因之隨機H3及3'部分。分別使用以下引子組合產生DP88-4文庫之此等文庫片段:片段1 (正向引子LMB3與反向引子Vk1_5_L3r_S或Vk1_5_L3r_SY或Vk1_5_L3r_SPY之組合)、片段2 (正向引子RJH31與反向引子RJH32之組合)及片段3 (正向引子DP88-v4-4或DP88-v4-6或DP88-v4-8與反向引子fdseqlong之組合)。用於製造文庫片段之PCR參數為94℃下5分鐘初始變性,1分鐘94℃,1分鐘58℃,1分鐘72℃的25次循環及在72℃下10分鐘的末端延長。對於裝配PCR,使用等莫耳比的凝膠純化之單一片段作為模板,參數為在94℃下3分鐘初始變性及30秒94℃、1分鐘58℃、2分鐘72℃的5次循環。在此階段,添加外部引子(LMB3及fdseqlong)且進行額外20次循環,隨後在72℃下末端延長10分鐘。裝配足夠量的全長隨機Fab構築體之後,將其用NcoI/NheI消化且接合至類似處理的受體噬菌粒載體中。經純化的連接用於約60次轉型至電感受態大腸桿菌TG1中。呈現Fab文庫之噬菌粒粒子補救且藉由PEG/NaCl純化進行純化,以供選擇。此等文庫建構步驟重複三次以獲得最終文庫尺寸4.4×10
9。如藉由點漬墨法中之C端標籤偵測所測定的功能性純系之百分比分別為輕鏈92.6%及重鏈93.7%。
對人類及獼猴4-1BB具有特異性之抗體
25G7選自Fab型式的通用噬菌體呈現抗體文庫(λ-DP47)。此文庫基於人類生殖系基因使用V結構域成對Vl3_19 (λ輕鏈)及VH3_23 (重鏈)建構,其包含輕鏈之CDR3 (L3,3種不同長度)及重鏈之CDR3 (H3,3種不同長度)中的隨機序列間隔。藉由重疊延長(SOE) PCR施加剪接,藉由3 PCR擴增之片段的組合件進行文庫繼代。片段1包含包括隨機L3之抗體基因的5'端,片段2為自L3跨越至H3的中部恆定片段而片段3包含抗體基因之隨機H3及3'部分。分別使用以下引子組合產生λ-DP47文庫之此等文庫片段:片段1 (正向引子LMB3與反向引子Vl_3_19_L3r_V或Vl_3_19_L3r_HV或Vl_3_19_L3r_HLV之組合)、片段2 (正向引子RJH80與反向引子MS63之組合)及片段3 (正向引子DP47-v4-4或DP47-v4-6或DP47-v4-8與反向引子fdseqlong之組合)。用於製造文庫片段之PCR參數為94℃下5分鐘初始變性,1分鐘94℃,1分鐘58℃,1分鐘72℃的25次循環及在72℃下10分鐘的末端延長。對於裝配PCR,使用等莫耳比的凝膠純化之單一片段作為模板,參數為在94℃下3分鐘初始變性及30秒94℃、1分鐘58℃、2分鐘72℃的5次循環。在此階段,添加外部引子(LMB3及fdseqlong)且進行額外20次循環,隨後在72℃下末端延長10分鐘。裝配足夠量的全長隨機Fab構築體之後,將其用NcoI/NheI消化且接合至類似處理的受體噬菌粒載體中。經純化的連接用於約60次轉型至電感受態大腸桿菌TG1中。呈現Fab文庫之噬菌粒粒子補救且藉由PEG/NaCl純化進行純化,以供選擇。獲得最終文庫尺寸9.5×10
9。如藉由點漬墨法中之C端標籤偵測所測定的功能性純系之百分比分別為輕鏈81.1%及重鏈83.2%。
表 38展示通用噬菌體呈現之抗體文庫(DP88-4)的序列,
表 39提供文庫DP88-4生殖系模板之cDNA及胺基酸序列且
表 40展示用於產生DP88-4生殖系模板的引子序列。
表 38 :通用噬菌體呈現抗體文庫(DP88-4)之序列
表 39 :文庫DP88-4生殖系模板之cDNA及胺基酸序列
表 40 :用於產生DP88-4文庫之引子序列
表 41展示用於PCR之通用噬菌體呈現λ-DP47文庫(Vl3_19/VH3_23)模板之序列。
表 42提供λ-DP47文庫(Vl3_19/VH3_23)生殖系模板之cDNA及胺基酸序列且
表 43展示用於產生λ-DP47文庫(Vl3_19/VH3_23)之引子序列。
表 41 :用於PCR之通用噬菌體呈現λ-DP47文庫(Vl3_19/VH3_23)模板之序列
表 42 :λ-DP47文庫(Vl3_19/VH3_23)生殖系模板之cDNA及胺基酸序列
表 43 :用於產生λ-DP47文庫(Vl3_19/VH3_23)之引子序列
1:G/D = 20%,E/V/S = 10%,A/P/R/L/T/Y=5%;2:G/Y/S=15%,A/D/T/R/P/L/V/N/W/F/I/E = 4,6%;3:G/A/Y = 20%,P/W/S/D/T = 8%;4: F = 46%,L/M = 15%,G/I/Y = 8%;5:K=70%,R =30%。
人類、鼠類及獼猴4-1BB (CD137)作為用於噬菌體呈現選擇以及基於ELISA及SPR之篩選的抗原,在HEK EBNA細胞中經短暫表現為N端單體Fc融合物且經由位於攜帶受體鏈(Fc杵鏈)之Fc部分的C端處之avi標籤識別序列處的BirA生物素接合酶之共表現發生活體內位點特異性生物素化。
根據以下程序,在溶液中進行數輪選擇(生物淘選)。第一步,在用10 μg/ml不相關人類IgG塗佈之maxisorp培養盤上預清理約10
12個噬菌粒粒子消耗識別抗原之Fc部分的抗體庫;第二,在100 nM不相關非生物素化Fc杵-臼構築體存在下用100 nM生物素化人類或鼠類4-1BB培育非結合噬菌粒粒子0.5小時,進一步消耗總體積1 ml之Fc結合子;第三,捕捉生物素化hu 4-1BB且藉由轉移至中性抗生物素蛋白預塗佈之微量滴定盤的4個孔中連接特異性結合噬菌體10分鐘(在第1輪及第3輪中);第四,使用5× PBS/Tween20及5× PBS洗滌各別孔;第五,藉由每孔添加250 μl 100 mM TEA (三乙胺)溶離噬菌粒粒子10分鐘且藉由向來自4個孔之彙聚溶離液添加500 μl 1M Tris/HCl pH 7.4中和;第六,藉由在中性抗生物素蛋白預塗佈之微量滴定盤上與100 nM生物素捕捉之Fc杵-臼構築體一起培育來後清理經中和之溶離液,最終移除Fc結合子;第七,用經溶離之噬菌粒粒子的清液層重新感染對數期大腸桿菌TG1細胞,用輔助噬菌體VCSM13感染,在30℃下在震盪器上培育隔夜且隨後PEG/NaCl沈澱打算用於後續選擇回合之噬菌粒粒子。使用100 nM之恆定抗原濃度進行3或4輪選擇。在第2輪及第4輪中,為避免中性抗生物素蛋白之結合子增濃,捕捉抗原:藉由添加5.4×10
7抗生蛋白鏈菌素塗佈之磁珠進行噬菌體複合。藉由ELISA如下鑑別特異性結合子:100 μl 25 nM生物素化人類或鼠類4-1BB及10 μg/ml人類IgG分別塗佈於中性抗生物素蛋白培養盤及maxisorp培養盤上。添加含Fab之細菌清液層且使用抗Flag/HRP二次抗體經由其Flag標籤偵測結合Fab。將在人類或鼠類4-1BB上展現信號且在人類IgG上為陰性的純系列入候選清單進行進一步分析且亦以類似方式針對其餘兩個物種之4-1BB進行測試。其在0.5公升培養體積中經細菌表現,進行親和力純化且使用BioRad之ProteOn XPR36生物感測器藉由SPR分析進行進一步表徵。
純系12B3、25G7、11D5及9B11經上文所述之程序識別為人類4-1BB特異性結合子。純系20G2經上文所述之程序識別為鼠類4-1BB特異性結合子。其可變區之cDNA序列展示於下
表 44中,相應胺基酸序列可見於表C中。
表 44 :源自噬菌體之抗-4-1BB抗體之可變區鹼基對序列。帶下劃線的是互補決定區(CDR)。
6.3 抗 - 4 - 1BB IgG1 P329G LALA 抗體之製備、純化及表徵在具有人類IgG1之恆定重鏈或恆定輕鏈之框架中次選殖所選擇之抗-4-1BB結合子之重鏈及輕鏈DNA序列之可變區。根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入杵及臼重鏈之恆定區以消除與Fcγ受體之結合。
抗-4-1BB純系之核苷酸及胺基酸序列展示於
表 45中。全部抗-4-1BB-Fc-融合編碼序列選殖至驅動來自MPSV啟動子之插入物的表現及含有位於CDS之3'端處的合成聚A信號序列的質體載體。此外,載體含有用於質體之游離型維護之EBV OriP序列。
表 45 : P329GLALA 人類 IgG1 型式中之抗 - 4 - 1BB 純系之序列
藉由使用聚伸乙基亞胺共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生所選擇之抗-4-BB抗體。細胞用相應表現載體以1:1比率(「載體重鏈」:「載體輕鏈」)轉染。
對於在500 mL搖瓶中製造,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。轉染細胞在210×g下離心5分鐘,且清液層置換為預溫熱之CD CHO培養基。在20 mL CD CHO培養基中混合表現載體(200 μg完全DNA)。在添加540 μL PEI之後,溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。在轉染之後一天,添加1 mM丙戊酸及具有補充劑之7% Feed。培養7天之後,藉由在210×g下離心15分鐘來收集清液層。溶液無菌過濾(0.22 μm過濾器),用疊氮化鈉補充至最終濃度0.01%(w/v),且保持於4℃下。
使用用於抗原Fc融合物之純化的如上文所描述之蛋白質A,藉由親和層析進行抗體分子自細胞培養物清液層之純化。
濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl溶液(pH 6.0)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
使用基於胺基酸序列計算之莫耳消光係數,藉由在280 nm型量測OD來測定經純化之抗體之蛋白質濃度。使用LabChipGXII (Caliper)在存在及不存在還原劑(Invitrogen, USA)之情況下藉由CE-SDS分析抗體之純度及分子量。在25℃下,使用在25 mM K
2HPO
4、125 mM NaCl、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析抗體樣品之聚集物含量。
表 46概述抗-4-BB P329G LALA IgG1抗體之產量及最終含量。
表 46 :抗-4-BB P329G LALA IgG1純系之生物化學分析
實例 7 抗 - 4 -BB 抗體之表徵 7.1 人類 4 - 1BB 上之結合 7.1.1 表面電漿子共振 ( 親合力 + 親和力 )藉由表面電漿子共振(SPR)評定源自噬菌體之4-1BB特異性抗體與重組4-1BB Fc(kih)之結合。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。
在同一實驗中,測定源自噬菌體呈現之抗-4-1BB純系12B3、25G7、11D5、9B11及20G2 (全部人類IgG1 P329GLALA)以及重組4-1BB (人類、獼猴及鼠類)之間的的物種選擇性及相互作用親合力。生物素化人類、獼猴及鼠類4-1BB Fc (kih)直接偶合至抗生蛋白鏈菌素(SA)感測器晶片之不同流槽。使用至多100個共振單位(RU)之固定程度。源自噬菌體呈現之抗-4-1BB人類IgG1 P329GLALA抗體在4至450 nM範圍內之濃度(3倍稀釋)下以30 μL/min之流速,經120秒通過流槽。監測複合物解離220秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。
使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導動力學常數,以藉由數值積分擬合1:1朗格繆爾結合之速率等式且用於定性評估親合力(
表 47)。
在同一實驗中,測定源自噬菌體呈現之抗體(人類IgG1 P329GLALA)與重組4-1BB(人類、獼猴及鼠類)之間的相互作用之親和力。抗人類Fab抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約7500 RU。在25 nM範圍內之濃度下捕捉針對4-1BB的源自噬菌體呈現之抗體持續60秒。重組人類4-1BB Fc (kih)以4.1至1000 nM範圍內之濃度,以30 μL/min之流速,經120秒通過流動槽。監測解離120秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fab抗體但已注射HBS-EP而非抗體之表面上流動。使用Biacore T200評估軟體(vAA,Biacore AB, Uppsala/Sweden)獲得動力學常數,以藉由數值積分針對1:1朗格繆爾結合來擬合比率等式。
純系25G7及9B11以低於純系12B3及11D5之親和力結合人類4-1BB Fc(kih)。純系20G2不結合於人類4-1BB。藉由與1:1朗格繆爾結合擬合來測定抗-4-1BB P329GLALA IgG1與人類4-1BB Fc (kih)之間的相互作用之親和力常數。
表 47 :抗-4-1BB抗體與重組人類4-1BB之結合
7.1.2 結合於人類 4 - 1BB 表現細胞 : 休眠及經活化之人類周邊單核血液白細胞 ( PBMC )休眠及原生人類T細胞上不存在人類4-1BB之表現(Kienzle G.及von Kempis J (2000), Int. Immunol. 12(1):, 73-82, Wen T.等人. (2002), J. Immunol. 168, 4897-4906)。用經固定之抗人類CD3促效抗體活化後,CD4
+及CD8
+T細胞上的4-1BB上調。亦報導經活化之人類NK細胞上之4-1BB表現(Baessler T.等人. (2010) Blood 115(15), 3058-3069)、經活化之人類NKT細胞上之4-1BB表現(Cole S.L.等人. (2014) J. Immunol. 192(8), 3898-3907)、經活化之人類B細胞上之4-1BB表現(Zhang等人. (2010) J. Immunol. 184(2), 787-795)、經活化之人類嗜酸性細胞上之4-1BB表現(Heinisch等人. 2001)、人類嗜中性白細胞上之組成性4-1BB表現(Heinisch I.V. (2000) J Allergy Clin Immunol. 108(1), 21-28)、經活化之人類單核細胞上之4-1BB表現(Langstein J.等人. (1998) J Immunol. 160(5), 2488-2494, Kwajah M.及Schwarz H. (2010) Eur J Immunol. 40(7), 1938-1949)、人類調節T細胞上之組成性4-1BB表現(Bacher P.等人. (2014) Mucosal Immunol. 7(4), 916-928)、人類濾泡樹突狀細胞上之4-1BB表現(Pauly S. 等人. (2002) J Leukoc Biol. 72(1), 35-42)、經活化之人類樹突狀細胞上之4-1BB表現(Zhang L.等人. (2004) Cell Mol Immunol. 1(1), 71-76)以及惡性人類腫瘤之血管上之4-1BB表現(Broll K.等人. (2001) Am J Clin Pathol. 115(4), 543-549)。
為了測試吾人抗-4-1BB純系對天然細胞表現之人類4-1BB的結合,使用新鮮分離之休眠周邊血液單核細胞(PBMC)或PHA-L/Proleukin預活化及CD3/CD28再活化PBMC。來自獲自Zurich血液供給中心之白血球層的PBMC使用Histopaque 1077 (SIGMA Life Science, 目錄號10771, 聚蔗糖及泛影酸鈉,調整至密度1.077 g/mL)藉由ficoll密度離心分離,且再懸浮於由具有10%胎牛血清(FBS, Gibco by Life Technology, 目錄號16000-044, 批號941273, γ照射,無支原體且在56℃下熱不活化35分鐘)、1% (v/v) GlutaMAX-I (GIBCO by Life Technologies, 目錄號35050 038)、1 mM丙酮酸鈉(SIGMA, 目錄號S8636)、1 % (v/v) MEM非必需胺基酸(SIGMA, 目錄號M7145)以及50 µM b-巰基乙醇(SIGMA, M3148)之RPMI 1640培養基(Gibco by Life Technology, 目錄號42401-042)組成的T細胞培養基中。PBMC在分離之後直接使用(休眠細胞)或經刺激以藉由在37℃及5% CO
2下,在6孔組織培養盤中在補充有200 U/mL Proleukin(Novartis Pharma Schweiz AG,CHCLB-P-476-700-10340)及2 μg/mL PHA-L (SIGMA,目錄號L2769)之T細胞中培養3至5天且接著在塗有含10 μg/mL抗人類CD3 (純系OKT3,BioLegend,目錄號317315)及2 μg/mL抗人類CD28 (純系CD28.2,BioLegend,目錄號:302928)之6孔組織培養盤中在T細胞培養基中培養2天來誘導4-1BB表現。
為了測定與人類PBMC表現之人類4-1BB之結合,向圓底懸浮細胞96孔培養盤(Greiner bio-one, cellstar, 目錄號650185)之各孔中添加0.1-0.2 × 10
6新鮮分離或活化之PBMC。培養盤用400×g且在4℃下離心4分鐘,且棄去清液層。細胞用200微升/孔DPBS洗滌,接著在4℃下與100 μL/mL含有1:5000倍稀釋之可固定活力染料eFluor 450 (eBioscience, 目錄號64-0863-18)或可固定的活力染料eFluor 660 (eBioscience, 目錄號65-0864-18)之DPBS一起培育30分鐘。隨後,細胞用200微升/孔冷FACS緩衝液(具有2% (v/v) FBS、5 mM EDTA pH8 (Amresco, 目錄號E177)及7.5 mM疊氮化鈉(Sigma-Aldrich S2002)之DPBS)洗滌一次。隨後,添加50 µL/孔含有經滴定之抗人類4-1BB結合子的4℃冷FACS緩衝液且在4℃下培育120分鐘。細胞用200 µL/孔4℃ FACS緩衝液洗滌四次,移除未經結合之分子。隨後,細胞與50 µL/孔含有2.5 μg/mL PE結合之AffiniPure抗人類IgG Fcγ-片段-特異性山羊F(ab`)2片段(Jackson ImmunoResearch, 目錄號109-116-098)或30 µg/mL FITC結合之AffiniPure抗人類IgG Fcγ-片段-特異性山羊F(ab`)2片段(Jackson ImmunoResearch, 目錄號109 096 098)、抗人類CD45 AF488 (純系HI30, BioLegend, 目錄號304019)、0.67 μg/mL APC/Cy7結合之抗人類CD3 moIgG1κ (純系UCH1, BioLegend, 目錄號300426)或0.125 μg/mL PE結合之抗人類CD3小鼠IgG1κ (純系SK7, BioLegend目錄號344806)或0.67 μg/mL PerCP/Cy5.5結合之抗人類CD3小鼠IgG1 κ (純系UCHT1, BioLegend, 目錄號300430)、0.125 μg/mL BV421結合之抗人類CD4 moIgG1κ (純系RPA-T4, BioLegend, 目錄號300532)或0.23 μg/mL BV421結合之抗人類CD4小鼠IgG2b κ (純系OKT4, BioLegend, 目錄號317434)或0.08 μg/mL PE/Cy7結合之抗人類CD4小鼠IgG1κ (純系SK3, BioLegend目錄號344612)、0.17 μg/mL APC/Cy7結合之抗人類CD8 (小鼠IgG1κ, 純系RPA-T8, BioLegend目錄號301016)或0.125 μg/mL PE/Cy7結合之抗人類CD8a (moIgG1κ, 純系RPA-T8, BioLegend,目錄號301012)或0.33 μg/mL抗人類CD8 BV510 (moIgG1κ, 純系SK1, BioLegend, 目錄號344732)及0.25 μg/mL APC結合之抗人類CD56 (小鼠IgG1κ, 純系HCD56, BioLegend, 目錄號318310)或1 μL AF488結合之抗人類CD56 (moIgG1κ, 純系B159, BD Pharmingen, 目錄號557699)及0.67 μg/mL抗人類CD19-PE/Cy7 (moIgG1κ, 純系HIB19, BioLegend, 目錄號302216)之4℃冷FACS緩衝液一起進一步培育且在4℃下培育30分鐘。
細胞用200 μL FACS緩衝液/孔洗滌兩次且藉由再懸浮於50 µL/孔含有1%甲醛(Sigma, HT501320-9.5L)之DPBS中固定。同一天或第二天使用3-雷射Canto II (BD Bioscience with DIVA software)或5-雷射Fortessa (BD Bioscience with DIVA software)或3-雷射MACSQuant分析器10 (Miltenyi Biotech)獲取細胞。閘設於CD8
+及CD4
+T細胞上且使用中位螢光強度(MFI)或二次偵測抗體之螢光強度的幾何平均值分析一次抗體之結合。使用Graph Pad Prism (Graph Pad Software Inc.),藉由減去空白值(未添加一次抗體)將資料基線化且使用非線性回歸曲線擬合(穩固擬合)計算EC50值。
人類T細胞在休眠狀態缺乏4-1BB表現,但活化之後4-1BB上調。人類CD8
+T細胞顯示比CD4
+T強的上調。。如
圖 23A - 23D 中所示,產生之抗人類4-1BB特異性抗體可結合於經活化之人類T細胞表現的人類4-1BB。所示之抗人類4-1BB純系可分類成強結合(純系12B3及11D5)及低結合子(純系25G7及9B11)。不僅EC
50值而且MFI都發現差異。與經活化之CD8 T細胞結合的EC50值顯示於
表 48中。抗小鼠4-1BB特異性純系20G2不結合於人類4-1BB且因此不具有人類交叉反應性。
表 48 :與經活化之人類 CD8 T 細胞結合的 EC
50 值
7.2 鼠類 4 - 1BB 上之結合 7.2.1 表面電漿子共振 ( 親合力 + 親和力 )藉由如上文針對人類4-1BB Fc (kih)所述的表面電漿子共振(參看實例7.1.1)評定源自噬菌體之4-1BB特異性抗體20G2與重組鼠類4-1BB Fc (kih)之結合。使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導動力學常數,以藉由數值積分擬合1:1朗格繆爾結合之速率等式且用於定性評估親合力(
表 44)。
對於親和力測定,由於Fc融合蛋白與參考流槽之非特異性相互作用,因此使用以AcTEV蛋白酶裂解之鼠類4-1BB Fc(kih)且藉由層析法移除Fc部分。抗人類Fc抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約7500 RU。在25 nM範圍內之濃度下捕捉針對4-1BB的源自噬菌體呈現之抗體持續60秒。重組鼠類4-1BB AcTEV以4.1至1000 nM範圍內之濃度,以30 μL/min之流速,經120秒通過流槽。監測解離120秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fc抗體但已注射HBS-EP而非抗體之表面上流動。
使用Biacore T200評估軟體(vAA,Biacore AB, Uppsala/Sweden)獲得動力學常數,以藉由數值積分針對1:1朗格繆爾結合來擬合比率等式。顯示純系20G2結合鼠類4-1BB (
表 49)。
抗-4-1BB P329GLALA IgG1分子與鼠類4-1BB之間的相互作用之親和力常數使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導,藉由數值積分擬合1:1朗格繆爾結合之速率等式。
表 49 :抗-4-1BB抗體20G2與鼠類4-1BB之結合
7.2.2 與小鼠 4 - 1BB 表現細胞之結合 : 休眠及經活化之小鼠脾細胞 ( 所選純系 )類似於人類,新鮮分離之休眠小鼠T細胞不表現4-1BB但表現可藉由TCR活化經肽脈衝之APC (Cannons J.等人. (2001) J. Immunol. 167(3): 1313-1324)或經固定之抗小鼠CD3或抗小鼠CD3與抗小鼠CD28抗體之組合(Pollok K.等人. (1995), European J. Immunol. 25(2), 488-494)誘導。經表面固定之抗CD3抗體活化之後,報導CD8
+T細胞上的4-1BB表現較高(Shuford W等人, (1997) J. of Experimental Med. 186(1), 47-55),但此可取決於活化方案。進一步4-1BB表現亦報導於經活化之小鼠NK細胞(Melero等人. (1998) Cell Immunol. 190(2), 167-172)、經活化之小鼠NKT細胞(Vinay D等人,. (2004) J Immunol. 173(6), 4218-4229)、經活化之小鼠B細胞(Vinay, Kwon (2011) Cell Mol Immunol. 8(4), 281-284)、小鼠嗜中性白細胞上之組成性表現(Lee S.等人(2005) Infect Immun. 73(8), 5144-5151)以及小鼠調節T細胞(Gavin等人. (2002) Nat Immunol. 3(1), 33-41)、小鼠IgE刺激肥大細胞(Nishimoto等人. (2005) Blood 106(13): 4241-4248)、小鼠骨髓系統細胞(Lee等人. (2008) Nat Immunol. 9(8), 917-926)、小鼠濾泡樹突狀細胞(Middendorp等人. (2009) Blood 114(11), 2280-2289)以及經活化之小鼠樹突狀細胞(Wilcox, Chapoval等人. (2002) J Immunol. 168(9),4262-4267)。
自健康雌性C57BL/6小鼠分離脾細胞之後以及用表面固定之促效抗小鼠CD3及抗小鼠CD28抗體活體外活化72小時之後,直接測試抗-4-1BB特異性抗體結合。雌性C57BL/6小鼠(年齡7-9週)購自Charles River, France。動物在到達之後維持一週以使其適應新環境且進行觀測。根據提交的指導原則(GV-Solas; Felasa; TierschG),小鼠保持在任意獲取食物及水且12小時光/12小時黑暗的日常循環的無特異性病原體之條件下。在常規基礎上進行連續健康監測。藉由頸椎脫位術處死小鼠。解剖脾臟且在補充有10% (v/v)加熱不活化FBS及1% (v/v) GlutaMAX-I之RPMI 1640中儲存於冰。為了獲得單細胞溶液,脾臟經70 µm細胞過濾器(BD Falcon; Germany)均質化且在37℃下在ACK溶解緩衝液(ddH
2O中0.15M NH4CL、10 mM KHCO3、0.1mM EDTA,pH 7.2)中溶血10分鐘。用無菌DPBS進行兩個洗滌步驟之後,將脾細胞在T細胞培養基中復原。任一細胞均新鮮使用(休眠)或10
6個細胞/mL脾細胞在由補充有10% FBS、1 % (v/v) GlutaMAX-I、1 mM丙酮酸鈉、1 % (v/v) MEM非必需胺基酸及50 µM β-巰基乙醇培養基組成之T細胞培養基中在塗佈有1 μg/mL抗小鼠CD3抗體(大鼠IgG2b,純系17A2, BioLegend,目錄號100223)及2 μg/mL抗小鼠CD28抗體(敍利亞倉鼠(syrian hamster), 純系37.51, BioLegend目錄號102112)之6孔細胞培養盤的RPMI 1640進一步刺激72小時。
為了測試與小鼠4-1BB之結合,新鮮分離或活化之小鼠脾細胞再懸浮於DPBS中且將0.1 × 10
6/孔脾細胞轉移至圓底96懸浮細胞培養盤(Greiner bio-one, cellstar,目錄號650185)中。細胞在4℃及400×g下離心4分鐘且移除清液層。再懸浮於100 μ/孔含有1:5000倍稀釋之可固定活力染料eFluor 450 (eBioscience, 目錄號65-0863-18)或可固定活力染料eFluor 660 (eBioscience, 目錄號65-0864-18)之DPBS中之後,細胞在4℃下培育30分鐘。細胞用FACS緩衝液及50微升/孔含有滴定濃度之抗人類4-1BB huIgG1 P329G LALA抗體-純系12B3、25G7、11D5、9B11及抗小鼠4-1BB-特異性純系20G2作為huIgG1 P329G LALA或小鼠IgG1或小鼠IgG1 DAPG的FACS緩衝液洗滌。在4℃下培育1小時之後,洗滌細胞四次移除過量抗體。若測試小鼠IgG1及小鼠IgG1 DAPG型式之抗小鼠20G2的結合,則細胞在4℃下在50 μL FACS緩衝液/孔含有30 μg/mL FITC結合之抗小鼠IgG Fcγ-片段-特異性AffiniPure山羊F(ab')γ片段中培育30分鐘且用FACS緩衝液洗滌兩次。若測試含有人類IgG1 P329G LALA Fc片段之抗-4-1BB結合子的結合,則跳過此步驟。隨後,細胞在含有0.67 μg/mL PE結合之抗小鼠CD3 (大鼠IgG2bκ, 純系17A2, BD Pharmingen, 目錄號555275)或APC-Cy7結合之抗小鼠CD3 (大鼠IgG2aκ, 純系53-6.7, BioLegend, 目錄號100708)、0.67 μg/mL PE/Cy7結合之抗小鼠CD4 (大鼠IgG2bκ, 純系GK1.5, BioLegend, 目錄號100422)、0.67 μg/mL APC/Cy7結合之抗小鼠CD8 (大鼠IgG2aκ, 純系53-6.7, BioLegend, 目錄號1007141)或PE結合之抗小鼠CD8 (大鼠IgG2aκ, 純系53-6.7, BioLegend, 目錄號100708)、2 μg/mL APC結合之抗小鼠NK1.1 (小鼠IgG2a, κ, 純系PK136, BioLegend, 目錄號108710)或PerCP/Cy5.5結合之抗小鼠NK1.1 (小鼠IgG2a, κ, 純系PK136, BioLegend, 目錄號108728)及10 μg/mL抗小鼠CD16/CD32 (小鼠Fc-Block, 大鼠IgG2b κ, 純系2.4G2, BD Bioscience, 目錄號553142)之50 μL FACS緩衝液/孔中培育。若測試含有人類IgG1 P329G LALA Fc片段之抗-4-1BB結合子之結合,則亦添加30 µg/mL FITC結合之AffiniPure抗人類IgG Fcγ-片段特異性山羊F(ab`)2片段(Jackson ImmunoResearch, 目錄號109 096 098)。在4℃下培育細胞30分鐘,用200微升/孔FACS緩衝液洗滌兩次且用50微升/孔含有1% (v/v)甲醛之DPBS再懸浮以供固定。第二天,細胞再懸浮於200微升/孔FACS緩衝液中且使用2-雷射CantoII (BD Bioscience with DIVA software)或5-雷射Fortessa (BD Bioscience with DIVA software)獲取。閘設於CD8
+及CD4
+T細胞上且使用二次檢測抗體之中位螢光強度(MFI)分析一次抗體之結合。使用Graph Pad Prism (Graph Pad Software Inc.),藉由減去空白值(未添加一次抗體)將資料基線化且使用非線性回歸曲線擬合(穩固擬合)計算EC
50值。
如
圖 24B 及 24D中所示,僅抗小鼠4-1BB結合純系20G2結合於經活化之小鼠CD8
+及CD4
+T細胞,而結合純系9B11、11D5、12B3及25G7之抗人類4-1BB不結合於小鼠4-1BB且因此不具有小鼠交叉反應性。僅來自測試純系之純系20G2可用作小鼠替代物。正如所料,抗-4-1BB結合純系均不顯示結合於新鮮分離之休眠小鼠T細胞(
圖 24A 及 24C)。類似於經活化之人類CD4
+T細胞,經活化之小鼠CD4
+T細胞表現4-1BB亦比經活化之小鼠CD8
+T細胞少。然而,差異不像人類T細胞一樣強,此亦可涉及不同活化方案。EC
50值展示於
表 50中。
表 50 : 與經活化之鼠類 CD8 及 CD4 T 細胞之結合的 EC
50 值
如
圖 25B 及 25D中所示,結合純系20G2之抗小鼠-4-1BB以類似方式結合於moIgG1野生型(wt)或moIgG1 DAPG型式的經活化之小鼠CD8
+及CD4
+T細胞。結合與
圖 24B 及 24D中所示之結合類似;因此,型式之改變不影響結合特性。吾人將純系20G2轉移至moIgG以防止在免疫勝任小鼠中觸發抗藥物抗體(ADA)。在人類IgG1構築體中,DAPG突變等效於P329G LALA突變,例如其防止經FcR
+免疫細胞交聯。EC
50值展示於
表 51中。
表 51 : 與經活化之鼠類 CD8 及 CD4 T 細胞之結合的 EC
50 值
7.3 獼猴 4 - 1BB 上之結合 7.3.1 表面電漿子共振 ( 親合力 + 親和力 )如上文針對人類4-1BB Fc(kih)(參看實例7.1.1)所述藉由表面電漿子共振(SPR)評定源自噬菌體之4-1BB特異性抗體12B3、25G7、11D5及9B11與重組獼猴4-1BB Fc(kih)之結合。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。使用Biacore T200評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導動力學常數,以藉由數值積分擬合1:1朗格繆爾結合之速率等式且用於定性評估親合力(
表 52)。
在同一實驗中,測定源自噬菌體呈現之抗體(人類IgG1 P329GLALA)與重組獼猴4-1BB Fc(kih)之間的相互作用之親和力。抗人類Fab抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約9000 RU。使用濃度25至100 nM捕捉源自噬菌體呈現之4-1BB的抗體。如針對人類4-1BB Fc(kih(實例7.1.1)所述進行實驗。
純系12B3、25G7、11D5及9B11以類似親和力結合獼猴4-1BB Fc(kih)(
表 52),但12B3及11D5以較高親合力結合於表現獼猴4-1BB之細胞。抗-4-1BB P329GLALA IgG1與獼猴4-1BB Fc(kih)之間的相互作用之親和力常數使用Biacore T100評估軟體(vAA, Biacore AB, Uppsala/Sweden)推導,藉由數值積分針對1:1朗格繆爾結合擬合速率等式。
表 52 :抗-4-1BB抗體與重組獼猴4-1BB Fc(kih)之結合
7.3.2 獼猴 4 - 1BB 表現細胞 : 經活化之獼猴周邊單核血液白細胞 ( PBMC ) 上的結合為了測試抗人類4-1BB結合純系與獼猴細胞之交叉反應性,如針對人類PBMC(
7 . 1 . 2)所述以微小差異使用密度梯度離心自肝素化血液分離健康食蟹猴(cynomolgus fascicularis)之PBMC。經分離之PBMC在1.5×10
6個細胞/mL細胞密度下在由具有10 % FBS、1 % (v/v) GlutaMAX-I、1 mM丙酮酸鈉、1 % (v/v) MEM非必需胺基酸及50 µM β-巰基乙醇之RPMI 1640培養基組成之T細胞培養基中在用10 μg/mL 抗-cyno-交叉反應性CD3 (mo IgG3 λ, 抗人類CD3, 純系SP34, BD Pharmingen, 目錄號556610)及2 μg/mL 抗-cyno-交叉反應性D28 (moIgG1κ, 抗人類CD28, 純系CD28.2, BioLegend, 目錄號140786)抗體塗佈的6孔細胞培養盤(Greiner Bio-One, Germany)上培養72小時。72小時後,採集刺激細胞且以0.1×10
6個細胞/孔之濃度接種至圓底懸浮細胞96孔培養盤(Greiner bio-one, cellstar, 目錄號650185)中。在4℃下,在50微升/孔含有不同濃度之一次抗人類4-1BB特異性huIgG P32G LALA抗體之FACS緩衝液中培育細胞2小時。隨後,細胞用200 μL/孔FACS緩衝液洗滌四次且在4℃下與50 μL/孔含有2 μL PE-結合之抗cyno-交叉反應性CD4 (moIgG2aκ, 抗人類CD4, 純系M-T477, BD Pharmingen, 目錄號556616)、1 μg/mL PerCP/Cy5.5-結合之抗-cyno-交叉反應性CD8 (moIgG1κ, 抗人類CD8, 純系RPA-T8, BioLegend, 目錄號301032)及30 µg/mL FITC結合之AffiniPure抗人類IgG Fcγ-片段-特異性山羊F(ab`)2片段(Jackson ImmunoResearch, 目錄號109 096 098)之FACS緩衝液一起進一步培育30分鐘。細胞用FACS緩衝液洗滌兩次且再懸浮於100微升/孔具有0.2 μg/mL之FACS緩衝液中來區別死細胞與活細胞。使用5-雷射Fortessa (BD Bioscience with DIVA software)立即獲取細胞。閘設於CD8
+及CD4
+T細胞上且使用二次檢測抗體之中位螢光強度(MFI)分析一次抗體之結合。使用Graph Pad Prism (Graph Pad Software Inc.),藉由減去空白值(未添加一次抗體)將資料基線化且使用非線性回歸曲線擬合(穩固擬合)計算EC
50值。
如
圖 26A 及 26B中所示,至少三個抗人類4-1BB純系(即12B3、11D5及25G7)對活化CD4
+及CD8
+T細胞上表現之獼猴4-1BB亦具有交叉反應性。有趣的是,結合曲線看起來與人類4-1BB類似,例如全部測試構築體中12B3及11D5顯示最高MFI及最低EC
50值,而25G7為較低MFI及較高EC
50值的較弱結合子(
表 53)。純系9B11僅在最高濃度100 nM下結合。此與人類4-1BB之結合相反,其中純系9B11優於純系25G7。經活化之獼猴CD4
+及CD8
+T細胞上4-1BB表現量之其他差異類似於經活化之人類T細胞(例如CD4
+T細胞)表現比CD8
+T細胞少得多的4-1BB。
表 53 : 與經活化之獼猴 CD8 及 CD4 T 細胞的結合的 EC
50 值
7.4 配位體阻斷特性使用人類4-1BB配位體(R&D systems)測定4-1BB特異性人類IgG1 P329GLALA抗體分子干擾4-1BB/4-1BB-配位體相互作用的能力。類似地,使用鼠類4-1BB配位體(R&D systems)評定抗鼠類4-1BB特異性IgG1 P329GLALA抗體20G2之配位體阻斷特性。
人類或鼠類4-1BB配位體(R&D systems)使用標準胺偶合試劑盒(Biacore, Freiburg/Germany)在約1500 RU,pH 5.0下直接偶合至CM5晶片的兩個流槽。重組人類或鼠類4-1BB Fc(kih)以濃度500 nM以30 μL/min之流速經90秒穿過第二流槽。忽略解離且源自噬菌體的抗-4-1BB人類IgG1 P329GLALA以200 nM濃度以30 μL/min流速經90秒通過兩個流槽。監測解離60秒。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗體流經具有固定之人類或鼠類4-1BB配位體但上面已注射HBS-EP代替重組人類4-1BB Fc(kih)的表面。
源自噬菌體的純系25G7結合於人類4-1BB與其4-1BB配位體的複合物(
表 54 ,圖 27A)。因此,此抗體不與結合於人類4-1BB之配位體競爭且因此稱為「非配位體阻斷」。相反地,純系12B3、11D5及9B11不結合於與其配位體關聯的人類4-1BB且因此稱為「配位體阻斷」。鼠類替代物20G2不結合於與其配位體關聯的鼠類4-1BB且亦稱為「配位體阻斷」。
表 54 :表面電漿子共振測定之抗-4-1BB純系之配位體結合特性
7.5 抗原決定基表徵藉由表面電漿子共振表徵源自噬菌體之抗-4-1BB抗體識別的抗原決定基。首先,藉由表面電漿子共振評定抗體競爭結合於人類4-1BB之能力。其次,如上文所述藉由SPR評估抗-4-1BB抗體與人類及鼠類4-1BB之兩個不同結構域的結合。為此,已設計雜交4-1BB Fc(kih)融合蛋白。其含有人類或鼠類結構域的4-1BB胞外域。雜交4-1BB變異體融合至Fc(kih)之杵鏈,類似於用於噬菌體呈現選擇及上文所述之抗原Fc融合。此等實驗之目標為界定「抗原決定基框組」。競爭類似或重疊抗原決定基之抗體不能同時結合於4-1BB且屬於「抗原決定基框組」。可同時結合於4-1BB之抗-4-1BB抗體不共用抗原決定基或其部分體,且因此分至不同抗原決定基框組。
7.5.1 競爭結合 ( SPR )為了分析針對抗-4-1BB人類IgG1 P329GLALA之人類或鼠類受體的競技結合(
圖 28A - 28F),源自噬菌體之抗-4-1BB純系9B11及11D5分別以1400及2200 RU (pH 5.5)使用標準胺偶合試劑盒(Biacore, Freiburg/Germany)直接偶合至CM5晶片。重組人類4-1BB Fc(kih)以濃度200 nM以30 μL/min流速經180秒注射。忽略解離且第二抗-4-1BB人類IgG1 P329GLALA抗體以濃度100 nM以流速30 μL/min經90秒通過。藉由減去參考流槽上所得之反應來校正整體折射率差異。
SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。競爭結合實驗顯示源自噬菌體之抗-4-1BB純系12B3、11D5及9B11與25G7共用不同空間抗原決定基,因為兩個抗體可同時結合於人類4-1BB Fc(kih) (
表 55)。
表 55 :競爭結合實驗之概述
O = 不結合 ; 1 , 結合 ; ; X = 未測定,因為 第二注射含有與固定於晶片上之抗體相同的抗體 7.5.2 與受體變異體 ( SPR ) 之結合源自噬菌體之抗-4-1BB純系12B3、25G7、11D5、9B11及20G2識別之抗原決定基已使用含有人類及鼠類結構域之組合或單個結構域的4-1BB Fc(kih)融合分子之雜交變異體表徵(
表 56)。如實例6.1中針對4-1BB Fc(kih)抗原所述,雜交4-1BB稠合至Fc(kih)。
表 52含有用於製備雜交4-1BB變異體之4-1BB結構域的序列。
構築體1由融合至人類4-1BB之C端部分(稱為結構域2)的鼠類4-1BB之N端部分(稱為結構域1)構成(
表 56)。構築體2由融合至鼠類4-1BB之C端部分(稱為結構域2)的人類4-1BB之N端部分(稱為結構域1)構成(
表 57)。因為結構域2之表現不會產生功能性分子,所以製備此等雜交分子。人類4-1BB之N端部分(結構域1)亦表現為Fc(kih)融合分子(構築體3)。如實例6.1中所述進行雜交4-1BB Fc(kih)變異體之表現及純化。
表 56 :雜交4-1BB Fc(kih)分子之胺基酸序列
表 57 :用於製備雜交構築體之人類及鼠類4-1BB結構域之胺基酸序列
SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。抗人類Fab抗體(Biacore, Freiburg/Germany)使用標準胺偶合套組(Biacore, Freiburg/Germany)在pH 5.0下在CM5晶片上直接偶合。固定含量為約8,000 RU。在100 nM下捕捉源自噬菌體呈現之抗-4-1BB抗體60秒。重組雜交人類/小鼠4-1BB Fc(kih)變異體在200 nM下以流速30 μL/min經120秒穿過流槽。監測解離120秒(
圖 29A - 29D)。
人類4-1BB-D1 (構築體3)以約1000 RU直接偶合至CM5晶片。200 nM下之抗-4-1BB抗體以30 μL/min持續180秒通過且監測解離60秒(
圖 30A - 30D)。藉由減去參考流槽上所得之反應來校正整體折射率差異。此處,抗原在具有固定之抗人類Fab抗體但已注射HBS-EP而非抗體之表面上流動。
源自噬菌體之純系12B3、11D5及9B11結合兩個含有人類結構域1之4-1BB Fc(kih)分子。源自噬菌體之純系25G7結合含有人類結構域2之雜交4-1BB Fc(kih)分子。源自噬菌體之純系20G2結合含有鼠類結構域1之雜交4-1BB Fc(kih)分子。概述可存在於
表 58中。
表 58 :雜交4-1BB Fc(kih)變異體與抗-4-1BB抗體之結合
1 = 結合; 0 = 未結合。
實例 8 抗 4 - 1BB 結合純系之功能特性 8.1 抗人類 4 - 1BB 結合純系之功能特性4-1BB用作共刺激受體且以TCR依賴性方式改良T細胞之擴展、細胞激素產生及功能特性。經表面固定之抗CD3抗體或肽脈衝之抗原呈現細胞的TCR活化體誘發T細胞上之4-1BB上調 (Pollok等人. (1993) J. Immunol. 150(3): 771-781)且藉由強化擴展及細胞激素釋放在嚙合後支持免疫反應(Hurtado等人. (1995) J Immunology 155(7), 3360-3367)。為了測試所產生之抗人類4-1BB抗體之強化能力,圓底懸浮96孔培養盤(Greiner bio-one, cellstar, 目錄號650185)用含有2 μg/mL AffiniPure F(ab')2片段山羊抗人類IgG, Fcγ-片段特異性(Jackson Immunoresearch, 目錄號109-006-008)及2 μg/mL AffiniPure山羊抗小鼠IgG, Fcγ-片段特異性(Jackson Immunoresearch, 目錄號115-005-008)之DPBS塗佈隔夜。培養盤用DPBS洗滌以移除過量分子且在37℃下用含有1% (w/v) BSA之DPBS (SIGMA-Aldrich, 目錄號A3059-100G)阻斷90分鐘。移除清液層且培養盤與具有1% (w/v) BSA且具有或不具有10 ng/mL抗人類CD3抗體之DPBS (BioLegend, 目錄號317315, 純系OKT3)一起在37℃下培育90分鐘。培養盤用DPBS洗滌且與具有1% (w/v)BSA及不同濃度之經滴定抗人類4-1BB IgG1 P329 G LALA抗體之DPBS一起培育。培養盤用DPBS洗滌且抽吸清液層。
人類PBMC如上文所述(
7 . 1 . 2)分離且在含有10 % (v/v) FBS、1% (v/v) GlutaMAX-I、2 μg/mL PHA-L及200 U/mL Proleukin之RPMI 1640中活化5天。細胞以1-2×10
6個細胞/毫升之密度在含有10 % (v/v) FBS、1% (v/v) GlutaMAX-I及200 U/mL Proleukin之RPMI 1640中進一步培養21天。採集、洗滌長期培養之PBMC,且根據製造商方案使用人類CD8
+T細胞分離試劑盒(Miltenyi Biotec, 目錄號130-096-495)分離CD8 T細胞。預活化及分選之CD8
+T細胞在200 μL/孔由具有10 % FBS、1 % (v/v) GlutaMAX-I、1 mM丙酮酸鈉、1 % (v/v) MEM非必需胺基酸及50 µM β-巰基乙醇之RPMI 1640培養基組成之T細胞培養基中接種達到7×10
4個細胞/孔。細胞培育72小時,最後4小時有Golgi-Stop (BD Bioscience, 目錄號554724)存在。細胞用DPBS洗滌且在4℃下在100 μL/孔含有1:5000倍稀釋之LIVE/DEAD可固定綠色死細胞染色劑之DPBS(Molecular Probes, Life Technologies, 目錄號L-23101)中培育30分鐘。隨後,洗滌細胞且在4℃下在50微升/孔含有0.5 μg/mL PerCP/Cy5.5結合之抗人類CD8 (小鼠IgG1 κ, 純系RPA-T8, BioLegend, 目錄號301032)、0.5 μg/mL PE/Cy7-結合之抗人類CD25 (小鼠IgG1 κ, 純系BC96, BioLegend, 目錄號302612)及1 μg/mL APC/Cy7-結合之抗人類PD-1 (小鼠IgG1 κ, 純系EH12.2H7, BioLegend, 目錄號329922)之FACS緩衝液中培育30分鐘。細胞用FACS緩衝液洗滌且以50微升/孔再懸浮於新鮮製備之固定/滲透溶液(eBioscience, 目錄號00-5523-00)中。在4℃下培育30分鐘之後,細胞用新鮮製備之滲透緩衝液(eBioscience, 目錄號00-5523-00)洗滌且在4℃下與50 μL/孔含有2 μg/mL APC標記之抗人類IFNγ (mo IgG1 κ, 純系B27, BD Pharmingen, 目錄號554702)及2 μg/mL PE結合之抗人類TNFα (mo IgG1 κ, 純系MAb11, BD Pharmingen, 目錄號554513)之Perm-緩衝液一起培育1小時。細胞用含有1%甲醛之DPBS洗滌及固定。第二天使用2-雷射Canto II (BD,DIVA軟件)獲取細胞。閘設於CD8
+T細胞上且測定TNFα及IFNγ分泌CD8
+T細胞之頻率。使用Graph Pad Prism (Graph Pad Software Inc.),對資料進行墨點分析且使用非線性消退曲線擬合(穩固擬合)計算曲線。
如上文所述,在無TCR或CD3刺激存在下,4-1BB嚙合對CD8
+T細胞功能(Pollok 1995)無作用,而在次佳CD3活化存在下,4-1BB之共刺激提高細胞激素分泌(
圖 31A 或 31C)。經CD3抗體次佳活化誘發總CD8
+T細胞群體中30% CD8
+T細胞中之IFNγ分泌。添加4-1BB共刺激以濃度依賴性方式將IFNγ
+CD8 T細胞群體增加至高達55%(
圖 31B)。
表 59展示相應EC
50值。總CD8
+T細胞群體之TNFα分泌可自23%增加至39%(
圖 31D)。類似於其結合特性,純系12B3及11D5對INFγ表現之提高的頻率及EC
50優於25G7及9B11。因此,吾人產生之純系為功能性的且可提高TCR介導之T細胞活化及功能。若抗-4-1BB特異性抗體未經表面固定,則其不提高CD8
+T細胞活化(未示出)。
表 59 :經活化之CD8
+T細胞中IFNγ分泌的提高的EC
50值
8.2 抗小鼠 4 - 1BB 結合純系 20G2 之活體外功能特性為了預防促效抗-4-1BB抗體經Fc受體交聯,向抗體之Fc部分中引入突變。類似於人類IgG1抗體中的P329G LALA突變,向小鼠IgG1分子之Fc部分中引入DAPG突變。為了測試此是否防止FcR交聯且因此活化能力,小鼠脾細胞用次佳濃度之抗小鼠CD3抗體及不同濃度之小鼠IgG1及小鼠IgG1 DAPG型式的純系20G2活化。
自健康雌性C57BL/6小鼠(年齡7-9週,Charles River, France)獲得脾臟且在3 mL MACS緩衝液中使用C試管(Miltenyi Biotec,目錄號. 130-096-334)及溫和MACS Octo解離劑(Miltenyi Biotec, 目錄號130-095-937)均質化。離心(400xg, 8min, RT)後,將細胞糰粒再懸浮於7 ml ACK溶解緩衝液中且在37℃下溶血10分鐘。藉由添加由具有10 % FBS、1 % (v/v) GlutaMAX-I、1 mM丙酮酸鈉、1 % (v/v) MEM非必需胺基酸及50 µM β-巰基乙醇之RPMI 1640培養基組成之T細胞培養基停止溶血作用。脾細胞用DPBS洗滌且經70 μm細胞過濾器(Corning目錄號431751)過濾。脾細胞在37℃ DPBS中再懸浮至2×10
7個細胞/毫升且添加細胞增殖染料eFluor 670 (eBioscience, 目錄號65-0840-90)達到最終濃度2.5 mM。細胞在37℃下培育10分鐘,藉由添加FBS停止標記法且洗滌細胞。200 μL/孔含有0.5 μg/mL抗小鼠CD3特異性倉鼠IgG (純系145-2C11, BD Bioscience, 目錄號553057)、0.01-50 nM經滴定之抗小鼠4-1BB特異性純系20G2小鼠IgG1或小鼠IgG1 DAPG或同型對照MOPC-21小鼠IgG1 (BD Bioscience, 目錄號554121)或同型對照DP47 moIgG1 DAPG之T細胞培養基及1×10
5脾細胞轉移至96孔圓底組織培養盤(TPP, 目錄號92097)的各孔中。培育培養盤48小時。洗滌細胞,再懸浮於25 μL/孔含有1 μg/mL BV711結合之抗小鼠CD8大鼠IgG2a (純系53-6.7, BioLegend, 目錄號100748)及2 μg/mL BV421結合之抗小鼠CD4大鼠IgG2a (純系RM4-5, BioLegend, 目錄號100544)之FACS緩衝液中且在4℃下培育20分鐘。細胞洗滌且再懸浮於100微升/孔新鮮製備之固定/滲透溶液(eBioscience, 目錄號00-5523-00)中。在4℃下培育30分鐘後,細胞用DPBS洗滌且再懸浮於100微升/孔含有10 μg/mL Alexa Fluor 488結合之抗小鼠Eomes大鼠IgG2a (純系DAN11MAG, eBioscience, 目錄號53-4875-82)及2 μg/mL PE結合之抗小鼠顆粒酶B大鼠IgG2a (純系NGZB, eBioscience, 目錄號12-8898-82)的新鮮製備之滲透緩衝液(eBioscience, 目錄號00-5523-00)中。細胞在室溫下在暗處培育1小時,用滲透緩衝液洗滌,再懸浮於FACS緩衝液中且使用5-雷射Fortessa (BD Bioscience, DIVA software)獲取。閘設於CD8
+T細胞上且測定Eomes及顆粒酶B表現CD8
+T細胞之頻率。使用Graph Pad Prism (Graph Pad Software Inc.),對資料進行墨點分析且使用非線性消退曲線擬合(穩固擬合)計算曲線。
CD8
+T細胞之最佳活化誘發脫中胚蛋白(Eomes)表現、調節CD8
+T細胞之細胞溶解效應機制的T-box轉錄因子(Glimcher等人. 2004)以及細胞溶解酶,如顆粒酶B。如
圖 32A 及 32B中所示,所用0.5 μg/mL抗小鼠CD3抗體濃度為次佳的且顆粒酶B表現CD8
+T細胞之頻率不超過30%且脫中胚蛋白(Eomes)表現CD8+ T細胞不超過18%。僅當添加足夠量之抗小鼠4-1BB純系20G2小鼠IgG1時,更多CD8
+T細胞開始表現顆粒酶B (
圖 32A)及脫中胚蛋白(
圖 32B)。此證實純系20G2為促效結合子且能夠藉由嚙合小鼠4-1BB活化T細胞。若純系20G2之交聯由於DAPG突變而減弱,則未發現改良之活化作用,此顯示亦需要抗-4-1BB抗體之小鼠細胞交聯來誘發4-1BB下游信號傳導。因此,DAPG突變防止充分交聯且經Fc結合受體呈現抗體。非小鼠-4-1BB特異性同型對照對T細胞活化不具作用。
8.3 抗小鼠 4 - 1BB 結合純系 20G2 之活體內功能特性用促效抗小鼠4-1BB大鼠IgG2a處理小鼠導致無腫瘤(Dubrot等人. (2010) Cancer Immunol Immunother. 59(8), 1223-33;Niu等人. (2007) J Immunol. 178(7):4194-213)及荷瘤小鼠(Wang等人. (2010) J Immunol. 185(12):7654-62; Kocak等人. (2006) Cancer Res. 66(14):7276-84)的肝臟中經活化T細胞之積聚且誘發肝炎。類似地,IV期黑素瘤患者中使用抗-4-1BB抗體Urelumab(huIgG4)之臨床II期試驗因為4級肝炎的罕見高發生率而必須終止(Ascierto P.等人. (2010) Semin Oncol. 37(5), 508-16)。為了測試抗小鼠4-1BB小鼠IgG1是否可誘發與所報導之促效抗小鼠4-1BB大鼠IgG2a抗體類似的肝臟中T細胞積聚,以及另外DAPG突變是否可預防肝臟中T細胞積聚,原生健康雌性C57BL/6小鼠(年齡7-9週,Charles River, France)用抗小鼠4-1BB IgG1及IgG DAPG純系20G2處理。小鼠如上文所述飼養及圈養。每週將150 μg抗體靜脈內注射至尾靜脈,持續3週。第二次注射後一天以及第三次注射之後一天及八天藉由頸椎脫位術處死小鼠。解剖脾臟及肝臟且在冰上儲存於補充有10% (v/v)熱不活化FBS及1% (v/v)GlutaMAX-I之RPMI 1640中。如上文(
8 . 2)所述分離脾細胞。如下自肝臟分離白血球:將各肝臟轉移至含有具有0.3 mg/mL分散酶II (Roche Applied Science, 目錄號:04942078001)、1 μg/mL Dnase I (Roche Applied Science, 目錄號:11284932001)及2 mg/mL膠原蛋白酶D (Roche Applied Science, 目錄號:11 088 882001)C管(Miltenyi Biotec, 目錄號130-096-334)之5 mL RPMI 1640,使用溫和MACS Octo解離劑使用程式「m_liver_01」 (Miltenyi Biotec, 目錄號130-095-937)破壞組織且肝臟在37℃下培育30分鐘。經消化之肝臟使用MACS Octo解離劑使用程式「m_liver_02」破壞第二次。離心(400xg, 8min, RT)後,將細胞糰粒再懸浮於7 ml ACK溶解緩衝液中且在37℃下溶血10分鐘。藉由添加具有10 % FBS及1 % (v/v) GlutaMAX-I之RPMI 1640培養基停止溶血作用。將10
6個細胞/孔脾細胞或來自肝臟之白血球轉移至圓底96孔懸浮細胞培養盤(Greiner bio-one, cellstar,目錄號650185)中且用DPBS洗滌一次。細胞再懸浮於100 μL/孔含有1:5000倍稀釋之LIVE/DEAD可固定綠色死細胞染色劑試劑盒(Live Technologies目錄號L-23101)且在4℃下培育30分鐘。隨後,洗滌細胞且再懸浮於50 μL/孔含有6 μg/mL PE結合之抗小鼠CD137敍利亞倉鼠IgG (純系17B5, BioLegend, 目錄號106106)或敍利亞倉鼠同型對照(純系SHG-1, BioLegend, 目錄號402008)、2 μg/mL PerCP/Cy5.5結合之抗小鼠TCR-β亞美尼亞倉鼠IgG (純系H57-597, BioLegend, 目錄號109228)、5 μg/mL PE/Cy7結合之抗小鼠CD25大鼠IgG1 (純系PC61, BD Bioscience, 目錄號552880)或大鼠IgG1 λ同型對照(純系A110-1, BD Bioscience, 目錄號552869), 2 μg/mL Alexa Fluor 700標記之抗小鼠CD8a ratIgG2a κ (純系53-6.7, BD Bioscience, 目錄號557959)、0.25 μg/mL BV421結合之抗小鼠CD4大鼠igG2b κ (純系GK1.5, BioLegend, 目錄號100438)、4 μg/mL BV605結合之抗小鼠CD11c亞美尼亞倉鼠IgG (純系N418, BioLegend, 目錄號117333)或亞美尼亞倉鼠同型對照(純系HTK888, BioLegend, 目錄號400943)之FACS緩衝液中且在4℃下培育30分鐘。細胞洗滌且再懸浮於100微升/孔新鮮製備之固定/滲透溶液(eBioscience, 目錄號00-5523-00)中。在4℃下培育30分鐘後,細胞用新鮮製備之滲透緩衝液(eBioscience, 目錄號00-5523-00)洗滌且再懸浮於50 μL/孔含有0.2 μg/mL eF660結合之抗小鼠Ki67大鼠IgG2a κ (純系SolA15, eBioscience, 目錄號50-5698-82)之滲透緩衝液中。細胞在4℃下培育30分鐘,用滲透緩衝液洗滌兩次且用50 μL/孔含有1%甲醛之DPBS固定。第二天,使用5-雷射Fortessa (BD Bioscience, DIVA software)獲取細胞。使用FlowJo (FlowJo, LLC)分析資料,閘設於CD8
+或CD4
+T細胞上且測定CD137
+、CD25
+、CD11c
+及Ki67
+T細胞之頻率。使用Graph Pad Prism(Graph Pad Software Inc.)對資料進行墨點分析。
如
圖 33中所示,僅用抗小鼠4-1BB純系20G2小鼠IgG1處理小鼠在肝臟中的CD8
+T細胞上誘發積聚、增殖及增加的4-1BB (CD137)表現,而防止經FcR結合與DAPG突變交聯繞過CD8
+T細胞之活化。防止與DAPG突變FcR可防止肝臟中的CD8
+T細胞活化(
圖 33),但兩種分子皆顯示與T細胞上表現之小鼠CD137的類似結合特性(
圖 25B 及 25D),且因此以相同可能活化特性為特徵。
實例 9 靶向 4 - 1BB 及腫瘤相關抗原 ( TAA ) 之雙特異性二價抗體的製備、純化及表徵 9.1 產生靶向 4 - 1BB 及纖維母細胞活化蛋白 ( FAP ) 之雙特異性二價抗體 ( 2 + 2 型式 )製備對4-1BB及FAP具有二價結合之雙特異性促效4-1BB抗體。如國際專利申請案第2010/145792 A1號中所述,應用互換單抗技術來減少錯誤配對輕鏈之形成。
FAP結合子之產生及製備描述於WO 2012/020006 A2中,其以引用之方式併入本文中。
在此實例中,FAP黏合劑28H1之交叉Fab單元(VHCL)使用(G4S)
4連接體序列C端融合至抗-4-1BB huIgG1之重鏈。此重鏈融合與抗-4-1BB之輕鏈及相應FAP交叉輕鏈(VLCH1)共表現。根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入重鏈之恆定區以消除與Fcγ受體之結合。所得雙特異性二價構築體類似於
圖 34A中描繪者。
表60分別展示成熟雙特異性二價抗-4-1BB/抗FAP人類IgG1 P329GLALA抗體之核苷酸及胺基酸序列。
表 60 :雙特異性二價抗-
4 - 1BB/抗FAP人類IgG1 P329GLALA抗原結合分子之序列
全部基因在嵌合MPSV啟動子控制下短暫表現,該啟動子由MPSV核啟動子與CMV啟動子強化子片段之組合組成。表現載體亦含有針對含有EBNA(埃-巴二氏病毒核抗原)之宿主細胞中游離型複製之oriP區。
藉由使用聚乙烯亞胺共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生雙特異性抗-4-1BB/抗FAP構築體。細胞用相應表現載體以1:1:1比率(「載體重鏈」:「載體輕鏈1」:「載體輕鏈2」)轉染。
關於在500 ml搖瓶中製造,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。關於轉染,使細胞在210×g下離心5分鐘,且用預溫熱之CD CHO培養基置換清液層。表現載體於20 ml CD CHO培養基中混合至200 μg DNA之最終量。在添加540 μL PEI之後,溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。轉染一天後,添加1 mM丙戊酸及7%饋料。在培養7天之後,藉由在210×g下離心15分鐘來收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
使用如上文針對抗原Fc融合物及抗體之純化所述的蛋白質A,藉由親和層析進行雙特異性構築體自細胞培養物清液層之純化。
濃縮且過濾蛋白質,隨後裝載於用20 mM組胺酸、140 mM NaCl溶液(pH 6.0)平衡之HiLoad Superdex 200管柱(GE Healthcare)上。
藉由使用基於胺基酸序列計算之莫耳消光係數量測280 nm下的OD來測定經純化之雙特異性構築體的蛋白質濃度。使用LabChipGXII (Caliper)在有及無還原劑(Invitrogen, USA)存在下藉由CE-SDS分析雙特異性構築體之純度及分子量。在25℃下,使用在25 mM K
2HPO
4、125 mM NaCl、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析性尺寸排阻管柱(Tosoh)分析雙特異性構築體之聚集物含量(表61)。
表 61 :雙特異性二價抗-4-1BB/抗FAP IgG1 P329G LALA抗原結合分子之生物化學分析
9.2 靶向 4 - 1BB 及 FAP 之雙特異性二價抗體的結合 9.2.1 表面電漿子共振 ( 同時結合 )藉由表面電漿子共振(SPR)評定同時結合人類4-1BB Fc (kih)及人類FAP之能力。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。
生物素化人類4-1BB Fc (kih)與抗生蛋白鏈菌素(SA)感測器晶片之流動細胞直接偶合。使用至多1000個共振單位(RU)之固定程度。靶向4-1BB及FAP之雙特異性抗體在250 nM範圍內之濃度下以30 μL/min之流速經90秒穿過流槽且解離設為零秒。以250 nM之濃度,以30微升/分鐘之流速,經流槽經90秒注射人類FAP作為第二分析物(
圖 34B)。監測解離120秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。全部雙特異性構築體均可同時結合於人類4-1BB及人類FAP (
圖 35A - 35D)。
9.2.2 細胞上之結合 9.2.2.1 結合於人類 4 - 1BB 表現細胞 : 休眠及經活化之人類周邊單核血液白細胞 ( PBMC )分離人類PBMC,活化以誘發4-1BB且如
實例 7 . 1 . 2中所述用於測試抗體與人類4-1BB之結合特性。
為了顯示Fc-融合之FAP靶向不會影響抗-4-1BB特異性純系之結合或經PE結合之人類IgG Fc特異性山羊IgG F(ab`)的偵測,吾人測試FAP靶向或DP47非靶向2+2構築體與親本huIgG P329G LALA型式相比結合休眠人類CD4
+及CD8
+T細胞(
圖 38A - 38F)及經活化之人類CD4
+及CD8
+T細胞(
圖 39A - 39F)之能力。類似於
圖 23,FAP靶向或DP47靶向之2+2人類4-1BB特異性分子亦不結合於休眠T細胞(
圖 38A - 38F),而是主要結合於經活化之CD8
+T細胞(
圖 39 D 、 E 及 F)。轉化成FAP靶向或DP47非靶向2+2格式不會顯著改變與4-1BB表現T細胞的結合行為。可藉由使用Fc特異性多株二次抗體偵測來解釋差異(例如Fc融合可遮蔽抗原決定基及減少偵測)。供體之間的EC
50值視活化之後表現的4-1BB的量而變化。此解釋
表 62中所列的EC
50值相較於
表 48中所示之親本抗體的EC
50值的差異。結合於經活化之CD8
+T細胞的曲線下面積展示於
圖 40中。
表 62 : 與經活化之人類 CD8 T 細胞結合的 EC
50 值
9.2.2.2 與表現人類 FAP 之腫瘤細胞之結合對於細胞表面表現之纖維母細胞活化蛋白(FAP)的結合分析,使用NIH/3T3-huFAP純系19細胞株或人類黑素瘤細胞株WM-266-4 (ATCC CRL-1676)。藉由在CMV啟動子下用編碼人類FAP之表現pETR4921質體轉染小鼠胚胎纖維母細胞NIH/3T3細胞(ATCC CRL-1658)來產生NIH/3T3-huFAP純系19。在1.5 μg/mL嘌呤黴素(InvivoGen,目錄號:ant-pr-5)存在下維持細胞。2 × 10
5個FAP表現腫瘤細胞添加至圓底懸浮細胞96孔培養盤(greiner bio-one, cellstar, 目錄號650185)的各孔中。細胞用200 µL DPBS洗滌一次,再懸浮於100 µL/孔含有1:5000倍稀釋之可固定活力染料eFluor 450 (eBioscience, 目錄號65 0863 18)或可固定活力染料eFluor 660 (eBioscience, 目錄號65-0864-18)之4℃冷DPBS緩衝液中且在4℃下培育30分鐘。細胞用200 µL 4℃冷DPBS緩衝液洗滌一次,再懸浮於50 µL/孔含有不同滴定濃度之4-1BB特異性FAP靶向及非靶向抗體之4℃冷FACS緩衝液中,隨後在4℃下培育1小時。在以200微升/孔洗滌四次之後,細胞用50微升/孔含有2.5 µg/mL PE結合之AffiniPure抗人類IgG Fcγ片段特異性山羊F(ab')2片段(Jackson ImmunoResearch,目錄號109-116-098)或30 µg/mL FITC結合之AffiniPure抗人類IgG Fgγ片段特異性山羊F(ab')2片段(Jackson ImmunoResearch,目錄號109 096 098)之4℃冷FACS緩衝液在4℃下染色30分鐘。細胞用200 µL 4℃ FACS緩衝液洗滌兩次且細胞再懸浮與50 µL/孔含有1%甲醛之DPBS中以供固定。同一天或第二天,將細胞再懸浮於100 µL FACS緩衝液中且使用5-雷射LSR-Fortessa (BD Bioscience with DIVA software)或MACSQuant分析器10 (Miltenyi Biotec)獲取。
如
圖 41A - 41F中所示,全部測試之2+2 FAP靶向分子而非含有4-1BB結合純系25G7、11D5及12B3純系之DP47靶向或親本huIgG1 P293G LALA型式有效結合且結合於人類FAP表現細胞。因此,測試之FAP (28H1)靶向2+2構築體可特異性結合於FAP表現細胞。
圖 42中顯示NIH/3T3-huFAP結合曲線之曲線下面積(AUC)的概述。測試的型式及FAP結合純系在下文中表示為圖形符號的曲線。顯示全部測試之FAP靶向2+2格式展示類似AUC。
表 63 : 結合於 FAP 表現細胞株 NIH / 3T3 - huFAP 純系 19 及 WM - 266 - 4 之 EC
50 值 .
9.2.3 NF κ B 活化 9.2.3.1 產生表現人類 4 - 1BB 及 NF - κB 螢光素酶之 HeLa 細胞子宮頸癌瘤細胞株HeLa (ATCC CCL-2)在CMV啟動子及嘌呤黴素抗性基因之控制下用基於表現載體pETR10829之質體轉導,該質體含有人類4-1BB (Uniprot寄存號Q07011)之序列。細胞在補充有10% (v/v)FBS、1% (v/v)GlutaMAX-I及3 μg/mL嘌呤黴素之DMEM培養基中培養。
藉由流動式細胞測量術測試經4-1BB轉導之HeLa細胞之4-1BB表現:0.2×10
6個活細胞再懸浮於含有0.1 µg PerCP/Cy5.5結合之抗人類4-1BB小鼠IgG1κ純系4B4-1 (BioLegend目錄號309814)或其同型對照物(PerCP/Cy5.5結合之小鼠IgG1κ同型對照抗體純系MOPC-21,BioLegend目錄號400150)之100 μL FACS緩衝液中且在4℃下培育30分鐘。細胞用FACS緩衝液洗滌兩次,再懸浮於含有0.06 μg DAPI (Santa Cruz Biotec,目錄號Sc-3598)之300 μL FACS緩衝液中且使用5-雷射LSR-Fortessa (BD Bioscience, DIVA software)獲取。如所描述進行有限稀釋以產生單一純系:經人類4-1BB轉導之HeLa細胞再懸浮於培養基中達到10、5及2.5個細胞/毫升之密度且將200 μl細胞懸浮液轉移至經組織培養物處理之圓底96孔培養盤(6個培養盤/細胞濃度,TPP目錄號92697)中。收集單一純系,擴增且如上文所描述測試4-1BB表現。選擇具有最高4-1BB 表現之純系(純系5)用於隨後用NF-κB螢光素酶表現載體5495p Tranlucent HygB進行之轉染。載體賦予經轉染細胞對濕黴素B之耐藥性及在NF-kB反應元素(Panomics, 目錄號LR0051)之控制下表現螢光素酶之能力。為了轉染,將人類-4-1BB HeLa純系5細胞培養至70%彙聚。將50 μg (40 μL)線性化(限制酶AseI及SalI) 5495p Tranlucent HygB表現載體添加至無菌0.4 cm Gene Pulser/MicroPulser Cuvette (Biorad, 目錄號165-2081)。添加含2.5×10
6個人類4-1BB HeLa純系5細胞之400 μl不含DMEM培養基之補充劑且與質體溶液小心混合。使用Gene Pulser Xcell完全系統(Biorad,目錄號165 2660)在以下設置下進行細胞之轉染:指數脈衝,電容500 μF,電壓160 V,電阻∞。在脈衝之後立即將經轉染之細胞轉移至具有15 mL 37℃溫熱的具有10% (v/v)FBS及1% (v/v)GlutaMAX-I之DMEM培養基的75 cm
2組織培養燒瓶(TPP,目錄號90075)中。第二天,添加含有3 μg/mL嘌呤黴素及200 μg/mL潮黴素B (Roche,目錄號10843555001)之培養基。擴增活細胞且如上文所描述進行有限稀釋以產生單一純系。
如上文所描述測試純系之4-1BB表現且如下測試NF-κB螢光素酶活性:在選擇培養基中收集純系且使用Cell Counter Vi-細胞xr 2.03 (Beckman Coulter,目錄號731050)計數。細胞設為細胞密度0.33×10
6個細胞/mL且將150 μL此細胞懸浮液轉移至具有蓋子的無菌白色96孔平底組織培養盤(greiner bio one, 目錄號655083)之各孔中。細胞在37℃及5% CO
2下在細胞培育箱(Hera Cell)中培育隔夜。第二天,50 μL含有不同濃度之重組型人類腫瘤壞死因子α (rhTNFα,PeproTech,目錄號300 01A)之培養基添加至96孔培養盤之各孔中達到100、50、25、12.5、6.25及0 ng/孔之rhTNFα之最終濃度。細胞在37℃及5% CO
2下培育6小時且接著用200微升/孔DPBS洗滌三次。報導體溶解緩衝液(Promega,目錄號:E3971)添加至各孔(40 μl)中且培養盤在-20℃下儲存隔夜。第二天,冷凍細胞培養盤及偵測緩衝液(Luciferase 1000分析系統,Promega,目錄號E4550)解凍至室溫。100 μL偵測緩衝液添加至各孔中且使用SpectraMax M5/M5e微板讀取器及SoftMax Pro軟體(Molecular Devices)儘可能快地量測盤。所量測之釋放光超過對照物(未添加rhTNFα)500毫秒/孔(URL)之單元視為螢光素酶活性。選擇呈現最高螢光素酶活性及大量4-1BB表現之HeLa-hu4-1BB-NF-κB-luc純系26用於進一步使用。
9.2.3.2 與表現 FAP 之 腫瘤細胞共培養之表現人類 4 - 1BB 報導體細胞 之 HeLa 細胞 中之 NFκB 活化採集HeLa-hu4-1BB-NF-κB-luc純系26細胞且再懸浮於具有10% (v/v) FBS及1% (v/v) GlutaMAX-I之DMEM培養基中達到0.2×10
6個細胞/毫升之濃度。此細胞懸浮液之100 μl (2×10
4個細胞)轉移至具有蓋子之無菌白色96孔平底組織培養盤(greiner bio-one,目錄號655083)之各孔中且培養盤在37℃及5% CO
2下培育隔夜。第二天,添加50 μL含有經滴定FAP靶向抗人類4-1BB構築體或其親本huIgG1 P329G LALA抗體之培養基。將FAP表現NIH/3T3-huFAP純系19或WM-266-4再懸浮於具有10 % (v/v) FBS及1 % (v/v) GlutaMAX-I之DMEM培養基中達到濃度2 × 10
6細胞/ml。
向各孔添加FAP表現腫瘤細胞之懸浮液(50 µl)或作為陰性對照之培養基(無FAP
+腫瘤細胞)且培養盤在37℃及5% CO
2下在細胞培育箱中培育6小時。細胞用200微升/孔DPBS洗滌兩次。40 µl新近製備之報導體溶解緩衝液(Promega,目錄號:E3971)添加至各孔中且盤在-20℃下儲存隔夜。第二天,在室溫下解凍冷凍細胞培養盤及偵測緩衝液(Luciferase 1000分析系統,Promega,目錄號E4550)。向各孔添加100 μL偵測緩衝液且儘快使用SpectraMax M5/M5e微量培養盤讀取器及SoftMax Pro軟體(Molecular Devices)量測螢光素酶活性。
FAP (28H1)靶向純系25G7、11D5或12B3 2+2構築體在NIH/3T3-huFAP細胞存在下(
圖 43 C 、 F 及 I)以濃度依賴性方式觸發報導體細胞株中的NFκB信號傳導路徑之活化。相比之下,未靶向之對照分子(4-1BB (25G7) × DP47 2+2)或親本huIgG1 P329G LALA抗體在任何測試濃度下均未能觸發此類作用。所測試之FAP靶向抗人類4-1BB 2+2抗體之此活性嚴格取決於所添加之交聯細胞的細胞表面處之FAP高表現,因為在無FAP表現腫瘤細胞存在下(
圖 43 A 、 D 及 G)或幾乎不表現FAP之腫瘤細胞株(WM-266-4)存在下(
圖 43 B 、 E 及 H)偵測不到NF-κB活化。因此,活化取決於FAP表現強度且不取決於抗4-1BB結合子之親和力強度,因為25G7 × FAP 2+2、11D5 × FAP 2+2及12B3 × FAP 2+2之間的活化效能(
圖 44A 及 44B 以及表 64)與其4-1BB結合親和力不相關(
圖 42)。
表 64 :在FAP表現NIH/3T3-huFAP純系19細胞存在下4-1BB表現報導體細胞株中NFκB控制之螢光素酶的活化的EC
50值
實例 10 靶向 4 - 1BB 及腫瘤相關抗原 ( TAA ) 之雙特異性單價抗體的製備、純化及表徵 10.1 產生單價型式 ( 1 + 1 型式 ) 的靶向 4 - 1BB 及纖維母細胞活化蛋白 ( FAP ) 的雙特異性抗體製備對4-1BB及FAP具有單價結合之雙特異性促效4-1BB抗體。如國際專利申請案第2010/145792 A1號中所述,應用互換單抗技術來減少錯誤配對輕鏈之形成。
FAP結合子之產生及製備描述於WO 2012/020006 A2中,其以引用之方式併入本文中。
雙特異性構築體單價結合於4-1BB及FAP (
圖 34C)。其含有與抗-4-1BB huIgG1之杵重鏈(含有S354C/T366W突變)融合的FAP結合子之交叉Fab單元(VHCL)。Fc臼重鏈 (含有Y349C/T366S/L368A/Y407V突變)融合至針對抗-4-1BB之Fab。靶向之抗FAP-Fc杵與抗4-1BB-Fc臼鏈之組合允許產生雜二聚體,其包括特異性結合於FAP之Fab及特異性結合於4-1BB之Fab。
根據國際專利申請公開案第WO 2012/130831 A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入杵及臼重鏈之恆定區以消除與Fcγ受體之結合。
使用聚乙烯亞胺藉由共轉染HEK293-EBNA細胞與哺乳動物表現載體來製造雙特異性單價抗-4-1BB及抗FAP huIgG1 P329GLALA。細胞用相應表現載體以1:1:1:1比率(「載體杵重鏈」:「載體輕鏈1」:「載體臼重鏈」:「載體輕鏈2」)轉染。
如針對雙特異性二價抗-4-1BB及抗FAP huIgG1 P329GLALA所述(參看實例9.1)製造及純化所得雙特異性單價構築體。核苷酸及胺基酸序列可見於
表 65中。
表 65 :成熟雙特異性單價抗-4-1BB/抗FAP huIgG1 P329GLALA kih抗體之cDNA及胺基酸序列
全部基因在嵌合MPSV啟動子控制下短暫表現,該啟動子由MPSV核啟動子與CMV啟動子強化子片段之組合組成。表現載體亦含有針對含有EBNA(埃-巴二氏病毒核抗原)之宿主細胞中游離型複製之oriP區。
藉由使用聚乙烯亞胺共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生雙特異性抗-4-1BB/抗FAP構築體。細胞用相應表現載體以1:1:1:1比率(「載體杵重鏈」:「載體臼重鏈」:「載體輕鏈1」:「載體輕鏈2」)轉染。
對於在500 mL搖瓶中製造,在轉染之前24小時接種400,000,000個HEK293 EBNA細胞。轉染細胞在210×g下離心5分鐘,且清液層置換為預溫熱之CD CHO培養基。表現載體於20 ml CD CHO培養基中混合至200 μg DNA之最終量。在添加540 μL PEI之後,溶液渦旋15秒且在室溫下培育10分鐘。隨後,將細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在具有5% CO
2氛圍的培育箱中在37℃下培育3小時。在培育之後,添加160 mL F17培養基且培養細胞24小時。轉染一天後,添加1 mM丙戊酸及7%饋料。在培養7天之後,藉由在210×g下離心15分鐘來收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
使用如上文針對抗原/Fc融合分子及抗體之純化所述的蛋白質A,藉由親和層析進行雙特異性構築體自細胞培養物清液層之純化。藉由使用基於胺基酸序列計算之莫耳消光係數量測280 nm下的OD來測定經純化之雙特異性構築體的蛋白質濃度。使用LabChipGXII (Caliper)在有及無還原劑(Invitrogen, USA)存在下藉由CE-SDS分析雙特異性構築體之純度及分子量。在25℃下,使用在25 mM K
2HPO
4、125 mM NaCl、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析雙特異性構築體之聚集物含量。
表 66 :例示性雙特異性單價抗-4-1BB/抗FAP IgG1 P329G LALA抗原結合分子之生物化學分析
10.2 作為篩選工具的 FAP 抗原之製備、純化及表徵為了測試與FAP之結合,編碼與C端HisTag融合之人類、小鼠或獼猴FAP之胞外域的DNA序列選殖至含有由MPSV核啟動子以及CMV啟動子強化片段組成之嵌合MPSV啟動子的表現載體中。表現載體亦含有針對含有EBNA(埃-巴二氏病毒核抗原)之宿主細胞中游離型複製之oriP區。His標記之人類FAP ECD之胺基酸及核苷酸序列分別展示於SEQ ID NO:85及86中。SEQ ID NO:88及89分別展示His標記之小鼠FAP ECD之胺基酸及核苷酸序列。SEQ ID NO:90及91分別展示His標記之獼猴FAP ECD之胺基酸及核苷酸序列。
藉由使用聚乙烯亞胺(PEI; Polysciences Inc.)共轉染HEK293-EBNA細胞與哺乳動物表現載體來產生FAP抗原。
對於500 mL搖瓶中的200 mL產量,300,000,000個HEK293 EBNA細胞接種24小時,隨後在100% F17 + 6mM麩醯胺酸中轉染。關於轉染,使400,000,000個細胞在210×g下離心5分鐘,且用20 mL預溫熱之CD CHO培養基(Gibco)置換清液層。表現載體於20 mL CD-CHO培養基中混合至200 μg DNA之最終量。在添加540 μL PEI (1 mg/mL) (Polysciences Inc.)之後,將溶液渦旋15秒且在室溫下培育10分鐘。隨後,再懸浮之細胞與DNA/PEI溶液混合,轉移至500 mL搖瓶中且在37℃下,在5% CO
2氛圍下且在165 rpm下振盪下,在培育箱中培育3小時。培育後,添加160 mL F17培養基及補充劑(1mM丙戊酸、5g/l Pepsoy及6 mM L-麩醯胺酸)且培養細胞24小時。轉染24小時後,細胞接著用12%最終體積(24 mL)的胺基酸及葡萄糖饋料補充。在培養7天之後,藉由在2000-3000×g下離心45分鐘來收集細胞清液層。將溶液無菌過濾(0.22 μm過濾器),補充疊氮化鈉至最終濃度為0.01% (w/v),且保持在4℃下。
以兩步驟純化自細胞培養物清液層純化抗原,首先使用5 ml IMAC管柱(Roche)或5 ml NiNTA管柱(Qiagen)之親和力層析步驟,隨後使用分別用20mM組胺酸、140 mM NaCl,pH6.0或2 mM MOPS 150 mM NaCl 0.02% NaN3 pH 7.3平衡之HiLoad Superdex 200管柱(GE Healthcare)的尺寸排阻層析。
對於使用IMAC管柱(Roche)之親和力層析法,用25 mM Tris-HCl、500 mM氯化鈉、20 mM咪唑,pH8.0平衡8 CV。裝載清液層且藉由用10 CV 25 mM Tris-HCl、500 mM氯化鈉、20 mM咪唑,pH8.0洗滌非結合之蛋白質。經結合之蛋白質使用線性梯度之20 CV (0-100% ) 25 mM Tris-HCl、500 mM氯化鈉、500 mM咪唑,pH8.0,隨後100%持續8 CV之步驟下溶離。
對於使用NiNTA管柱(Qiagen)之親和力層析,用50 mM磷酸鈉、300 mM氯化鈉,pH8.0平衡8 CV。裝載清液層且未結合之蛋白質藉由用10管柱體積之50 mM磷酸鈉、300 mM氯化鈉,pH8.0洗滌移除。結合之蛋白質使用線性梯度之20 CV (0至100%) 50 mM磷酸鈉、300 mM氯化鈉、500 mM咪唑,pH7.4,隨後5CV 50 mM磷酸鈉、300 mM氯化鈉、500 mM咪唑,pH7.4之步驟溶離。管柱接著用8 CV 50 mM磷酸鈉、300 mM氯化鈉,pH8.0重新平衡。
收集之溶離份接著用1/10 (v/v) 0.5 M EDTA,pH8.0補充。蛋白質在Vivaspin管柱(30 kD截止值,Sartorius)中濃縮,且在裝載於用2 mM MOPS 150 mM NaCl 0.02% NaN3 pH 7.3或20mM組胺酸, 140mM NaCl, pH6.0平衡的HiLoad Superdex 200管柱(GE Healthcare)上之前過濾。
藉由使用基於胺基酸序列計算之莫耳消光係數量測280 nm下的OD來測定經純化之抗原的蛋白質濃度。使用LabChipGXII (Caliper)在有及無還原劑(Invitrogen)存在下藉由CE-SDS分析抗原之純度及分子量。在25℃下,使用在25 mM磷酸鉀、125 mM氯化鈉、200 mM L-精胺酸單鹽酸鹽、0.02% (w/v)NaN
3、pH 6.7操作緩衝液中平衡之TSKgel G3000 SW XL分析型尺寸排阻管柱(Tosoh)分析抗原之聚集物含量。
10.3 靶向 4 - 1BB 及 FAP 之雙特異性單價抗體的結合 10.3.1 表面電漿子共振 ( 同時結合 )藉由表面電漿子共振(SPR)評定同時結合人類4-1BB Fc (kih)及人類FAP之能力。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20, Biacore, Freiburg/Germany)來進行。
生物素化人類4-1BB Fc (kih)與抗生蛋白鏈菌素(SA)感測器晶片之流動細胞直接偶合。使用至多1000個共振單位(RU)之固定程度。靶向4-1BB及FAP之雙特異性抗體在250 nM範圍內之濃度下以30 μL/min之流速經90秒穿過流槽且解離設為零秒。以250 nM之濃度,以30微升/分鐘之流速,經流槽經90秒注射人類FAP作為第二分析物。監測解離120秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。全部雙特異性構築體可同時結合人類4-1BB及人類FAP。
10.3.2 細胞上之結合 10.3.2.1 結合於人類 4 - 1BB 表現細胞 : 休眠及經活化之人類周邊單核血液白細胞 ( PBMC )分離人類PBMC,新鮮使用(例如休眠T細胞)或經活化以誘導4-1BB且用於如
實例 7 . 1 . 2中所述測試抗體與人類4-1BB之結合特性。
為了比較4-1BB FAP靶向1+1構築體之單價4-1BB結合與親本抗-4-1BB特異性huIgG1 P329G LALA純系之二價-4-1BB結合,構築體與休眠或經活化之人類4-1BB表現PBMC一起培育且如上文所述用PE結合之人類IgG Fc特異性山羊IgG F(ab`)偵測。如
圖 39D 及 39F中所示,轉化成單價4-1BB FAP靶向1+1型式相較於其親本huIgG1 P329G LALA構築體增加4-1BB結合之EC
50值(
表 67)且減小MFI值(
圖 39B / E 及 39C / F)。此EC
50增加且MFI見效導致如
圖 40中所示之結合曲線的曲線下面積(AUC)或多或少對等分。
表 67 :與經活化之人類 CD8 + T 細胞結合的 EC
50 值
10.3.2.2 與人類 FAP 表現細胞之結合與FAP表現腫瘤細胞之結合已描述於上文之
9 . 2 . 2 . 2中。
如
圖 41中所示,純系25G7及12B3之FAP靶向之1+1構築體而非DP47靶向之1+1或親本huIgG1 P293G LALA抗體有效結合至人類FAP表現細胞(
圖 41 B / D 及 41E / F)。因此,但由於以略高於FAP靶向2+2型式之EC
50單價FAP靶向,1+1構築體亦可特異性結合於FAP表現細胞(
表 68 及圖 41 B / D 及 41E / F)。然而,此對結合曲線之AUC僅具有最小作用(
圖 42)。
68 : 結合於 FAP 表現細胞株 NIH / 3T3 - huFAP 純系 19 及 WM - 266 - 4 之 EC
50 值
10.3.3. NF κ B 活化使用人類4-1BB表現報導體細胞株HeLa-hu4-1BB-NFκB-luc分析功能能力的方案已描述於
實例 9 . 2 . 3 . 2中。
FAP (28H1)-靶向純系25G7或12B3 1+1構築體在NIH/3T3-huFAP細胞存在下觸發報導體細胞株中NFκB信號傳導路徑之活化(
圖 43 F 及 I)。相比之下,未靶向之對照分子(4-1BB (25G7) × DP47 1+1)或親本huIgG1 P329G LALA抗體在任何測試濃度下均未能觸發此類作用。所測試之FAP靶向抗人類4-1BB 1+1抗體之此活性嚴格取決於所添加之FAP+細胞的細胞表面處之FAP高表現,因為在無FAP表現腫瘤細胞存在下(
圖 43 D 及 G)或幾乎不表現FAP之腫瘤細胞株(WM-266-4)存在下(
圖 43 E 及 H)偵測不到NF-κB活化。比較FAP靶向1+1與FAP靶向2+2分子純系12B3及25G7表現得不同。12B3 FAP靶向之1+1構築體相較於12B3 FAP靶向2+2構築體具有較高EC
50,而且活化平穩階段亦較高(
圖 43F),且因此兩種構築體具有類似AUC (
圖 44A),而25G7 FAP靶向1+1構築體相較於25G7 FAP靶向2+2構築體具有較高EC
50值,且活化平穩階段相同(
圖 44I),且因此較小AUC (
圖 44A),例如25G7 FAP靶向1+1明顯表現得比25G7 FAP靶向2+2差。原因可能是強4-1BB結合子(例如12B3)及低4-1BB結合子(例如25G7)之間的差異,此對於單價4-1BB結合可變的更明顯。
表 69 :在FAP表現腫瘤細胞存在下NFκB信號傳導路徑之活化的EC
50值
實例 11 二價結合於 4 - 1BB 且單價結合於腫瘤相關抗原 ( TAA ) 之雙特異性抗體的製備、純化及表徵 11.1. 二價結合於 4 - 1BB 且單價結合於腫瘤相關抗原 ( TAA ) 之雙特異性抗體 ( 2 + 1 型式 ) 的產生對4-1BB二價結合且對FAP單價結合之雙特異性促效4-1BB抗體(亦稱為2+1)已如
圖 36中所描繪製備。
在此實例中,構築體之第一重鏈HC1包含以下組分:抗4-1BB結合子之VHCH1,隨後Fc杵,在該C端融合有抗FAP結合子之VL或VH。第二重鏈HC2包含抗-4-1BB之VHCH1,隨後Fc臼,在該C端分別融合有抗FAP結合子(純系4B9)之VH或VL。FAP結合子4B9之產生及製備描述於WO 2012/020006 A2中,其以引用之方式併入本文中。如實例6中所述產生針對4-1BB(12B3、9B11、11D5及25G7)之結合子。靶向抗FAP-Fc杵與抗-4-1BB-Fc臼鏈之組合允許產生雜二聚體,其包括FAP結合部分及兩個4-1BB結合Fab (
圖 36A 及 36B)。
根據國際專利申請公開案第WO2012/130831A1號中描述之方法,將Pro329Gly、Leu234Ala及Leu235Ala突變引入杵及臼重鏈之恆定區以消除與Fcγ受體之結合。
使用聚乙烯亞胺藉由共轉染HEK293-EBNA細胞與哺乳動物表現載體來製造雙特異性2+1抗-4-1BB抗FAP huIgG1 P329GLALA抗體。細胞用相應表現載體以1:1:1比率(「載體杵重鏈」:「載體輕鏈」:「載體臼重鏈」)轉染。構築體如針對雙特異性二價抗-4-1BB及抗-FAP huIgG1 P329GLALA抗體所述產生及純化(參看實例9.1)。
2+1抗-4-1BB、抗FAP構築體之鹼基對及胺基酸序列可分別在
表 70中發現,其中構築體之a-FAP VH融合至杵且VL融合至臼鏈。
表 70 : 成熟雙特異性 2 + 1 抗 - 4 - 1BB 、抗 FAP 人類 IgG1 P329GLALA 之 cDNA 及胺基酸序列。 ( 構築體之 a - FAP V 融合至 臼且 VH 融合至 杵鏈 ,在下表 72 中稱為臼 - VL )
2+1抗-4-1BB、抗FAP構築體之鹼基對及胺基酸序列可分別在表71中發現,其中該等構築體之a-FAP VL融合至杵且VH融合至臼鏈。
表 71 : 成熟雙特異性 2 + 1 抗 - 4 - 1BB 、抗 FAP 人類 IgG1 P329GLALA 之 cDNA 及胺基酸序列。 ( 構築體之 a - FAP VH 融合至 臼且 VL 稠合至杵鏈 , 下文稱為臼 - VH )
表 72 : 二價結合於 4-1BB 且單價結合於 FAP (2+1 4-1BB/FAP 人類 IgG1 P329GLALA) 之雙特異性構築體的生物化學分析
11.2. 靶向 4 - 1BB 及 FAP 之雙特異性單價抗體的結合 11.2.1. 表面電漿子共振 ( 同時結合 )藉由表面電漿子共振(SPR)評定同時結合人類4-1BB Fc (kih)及人類FAP之能力。所有SPR實驗均在Biacore T200上在25℃下以HBS-EP作為操作緩衝液(0.01 M HEPES pH 7.4、0.15 M NaCl、3 mM EDTA、0.005%界面活性劑P20,Biacore, Freiburg/Germany)來進行。
生物素化人類4-1BB Fc (kih)與抗生蛋白鏈菌素(SA)感測器晶片之流動細胞直接偶合。使用至多400個共振單位(RU)之固定程度。靶向4-1BB及FAP之雙特異性抗體在200 nM範圍內之濃度下以30 μL/min之流速經90秒穿過流槽且解離設為零秒。以500 nM之濃度,以30微升/分鐘之流速,經流槽經90秒注射人類FAP作為第二分析物。監測解離120秒。藉由減去在參考流動細胞(其中未固定蛋白質)中獲得之反應來校正整體折射率差異。
如
圖 37B及
37C中所示,全部雙特異性構築體均可同時結合於人類4-1BB及人類FAP。
11.2.2 NF κ B 活化NFκB-報導體細胞株HeLa-huCD137-NFκB-luc純系26之產生以及活化分析法之設置已在9.2.3下描述。
含有純系11D5 (圖44A-C)或純系25G7 (圖G-I)之所測試之FAP (4B9)靶向2+1構築體在FAP表現腫瘤細胞存在下觸發報導體細胞株中NFκB信號傳導路徑之活化。此活性嚴格取決於腫瘤細胞之細胞表面處FAP的表現,因為在無FAP表現腫瘤細胞存在的情況下偵測不到NF-κB活化(
圖 43D 及 G)。與其他測試型式(例如2+2、1+1)不同,FAP (4B9)-靶向2+1亦可在較低FAP表現WM-266-4細胞存在下誘發活化(圖44E及H)。在NIH/3T3-huFAP純系19結合子存在下,2+1型式亦優於FAP (28H1)靶向2+2或1+1構築體(
圖 43F 及 I ,圖 44B)。此可使用較強FAP結合子(4B9>28H1)以及FAP結合側與4-1BB結合側之間的不同比率(1:2對比1:1)解釋。
表 73 :在FAP表現腫瘤細胞存在下NFκB信號傳導路徑之活化的EC
50值
***
CDR | Kabat | Chothia | AbM 2 |
V HCDR1 | 31-35 | 26-32 | 26-35 |
V HCDR2 | 50-65 | 52-58 | 50-58 |
V HCDR3 | 95-102 | 95-102 | 95-102 |
V LCDR1 | 24-34 | 26-32 | 24-34 |
V LCDR2 | 50-56 | 50-52 | 50-56 |
V LCDR3 | 89-97 | 91-96 | 89-97 |
初始殘基 | 例示性取代 | 較佳取代 |
Ala (A) | Val;Leu;Ile | Val |
Arg (R) | Lys;Gln;Asn | Lys |
Asn (N) | Gln;His;Asp, Lys;Arg | Gln |
Asp (D) | Glu;Asn | Glu |
Cys (C) | Ser;Ala | Ser |
Gln (Q) | Asn;Glu | Asn |
Glu (E) | Asp;Gln | Asp |
Gly (G) | Ala | Ala |
His (H) | Asn;Gln;Lys;Arg | Arg |
Ile (I) | Leu;Val;Met;Ala;Phe;正白胺酸 | Leu |
Leu (L) | 正白胺酸;Ile;Val;Met;Ala;Phe | Ile |
Lys (K) | Arg;Gln;Asn | Arg |
Met (M) | Leu;Phe;Ile | Leu |
Phe (F) | Trp;Leu;Val;Ile;Ala;Tyr | Tyr |
Pro (P) | Ala | Ala |
Ser (S) | Thr | Thr |
Thr (T) | Val;Ser | Ser |
Trp (W) | Tyr;Phe | Tyr |
Tyr (Y) | Trp;Phe;Thr;Ser | Phe |
Val (V) | Ile;Leu;Met;Phe;Ala;正白胺酸 | Leu |
SEQ ID NO: | 名稱 | 序列 |
1 | 人類OX40 ECD | Uniprot編號P43489, aa 29-214 |
2 | OX40(8H9,49B4,1G4, 20B7) CDR-H1 | SYAIS |
3 | OX40(CLC-563, CLC-564, 17A9) CDR-H1 | SYAMS |
4 | OX40(8H9,49B4,1G4, 20B7) CDR-H2 | GIIPIFGTANYAQKFQG |
5 | OX40(CLC-563, CLC-564, 17A9) CDR-H2 | AISGSGGSTYYADSVKG |
6 | OX40(8H9) CDR-H3 | EYGWMDY |
7 | OX40(49B4) CDR-H3 | EYYRGPYDY |
8 | OX40(1G4) CDR-H3 | EYGSMDY |
9 | OX40(20B7) CDR-H3 | VNYPYSYWGDFDY |
10 | OX40(CLC-563) CDR-H3 | DVGAFDY |
11 | OX40(CLC-564) CDR-H3 | DVGPFDY |
12 | OX40(17A9)-CDR-H3 | VFYRGGVSMDY |
13 | OX40(8H9,49B4,1G4, 20B7) CDR-L1 | RASQSISSWLA |
14 | OX40(CLC-563, CLC564) CDR-L1 | RASQSVSSSYLA |
15 | OX40(17A9) CDR-L1 | QGDSLRSYYAS |
16 | OX40(8H9,49B4,1G4, 20B7) CDR-L2 | DASSLES |
17 | OX40(CLC-563, CLC564) CDR-L2 | GASSRAT |
18 | OX40(17A9) CDR-L2 | GKNNRPS |
19 | OX40(8H9) CDR-L3 | QQYLTYSRFT |
20 | OX40(49B4) CDR-L3 | QQYSSQPYT |
21 | OX40(1G4) CDR-L3 | QQYISYSMLT |
22 | OX40(20B7) CDR-L3 | QQYQAFSLT |
23 | OX40(CLC-563, CLC-164) CDR-L3 | QQYGSSPLT |
24 | OX40(17A9) CDR-L3 | NSRVMPHNRV |
25 | OX40(8H9) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR EYGWMDYWGQGTTVTVSS |
26 | OX40(8H9) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYLTYSRFTFGQGTKVEIK |
27 | OX40(49B4) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR EYYRGPYDYWGQGTTVTVSS |
28 | OX40(49B4) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYSSQPYTFGQGTKVEIK |
29 | OX40(1G4) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR EYGSMDYWGQGTTVTVSS |
30 | OX40(1G4) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYISYSMLTFGQGTKVEIK |
31 | OX40(20B7) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR VNYPYSYWGDFDYWGQGTTVTVSS |
32 | OX40(20B7) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYQAFSLTFGQGTKVEIK |
33 | OX40(CLC-563) VH | EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRQAPGKGLEWVS AISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAL DVGAFDYWGQGALVTVSS |
34 | OX40(CLC-563) VL | EIVLTQSPGTLSLSPGERATLSC RASQSVSSSYLAWYQQKPGQAPRLLIY GASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQYGSSPLTFGQGTKVEIK |
35 | OX40(CLC-564) VH | EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRQAPGKGLEWVS AISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAF DVGPFDYWGQGTLVTVSS |
36 | OX40(CLC-564) VL | EIVLTQSPGTLSLSPGERATLSC RASQSVSSSYLAWYQQKPGQAPRLLIY GASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQYGSSPLTFGQGTKVEIK |
37 | OX40(17A9) VH | EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRQAPGKGLEWVS AISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR VFYRGGVSMDYWGQGTLVTVSS |
38 | OX40(17A9) VL | SSELTQDPAVSVALGQTVRITC QGDSLRSYYASWYQQKPGQAPVLVIY GKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSRVMPHNRVFGGGTKLTV |
39 | 人類4-1BB ECD | Uniprot編號Q07011, aa 24-186 |
40 | 4-1BB(12B3,11D5, 9B11, 20G2) CDR-H1 | SYAIS |
41 | 4-1BB(25G7) CDR-H1 | SYAMS |
42 | 4-1BB(12B3,11D5, 9B11, 20G2) CDR-H2 | GIIPIFGTANYAQKFQG |
43 | 4-1BB(25G7) CDR-H2 | AISGSGGSTYYADSVKG |
44 | 4-1BB(12B3) CDR-H3 | SEFRFYADFDY |
45 | 4-1BB(25G7) CDR-H3 | DDPWPPFDY |
46 | 4-1BB(11D5) CDR-H3 | STLIYGYFDY |
47 | 4-1BB(9B11) CDR-H3 | SSGAYPGYFDY |
48 | 4-1BB(20G2) CDR-H3 | SYYWESYPFDY |
49 | 4-1BB(12B3,11D5, 9B11, 20G2) CDR-L1 | RASQSISSWLA |
50 | 4-1BB(25G7) CDR-L1 | QGDSLRSYYAS |
51 | 4-1BB(12B3,11D5, 9B11, 20G2) CDR-L2 | DASSLES |
52 | 4-1BB(25G7) CDR-L2 | GKNNRPS |
53 | 4-1BB(12B3) CDR-L3 | QQYHSYPQT |
54 | 4-1BB(25G7) CDR-L3 | NSLDRRGMWV |
55 | 4-1BB(11D5) CDR-L3 | QQLNSYPQT |
56 | 4-1BB(9B11) CDR-L3 | QQVNSYPQT |
57 | 4-1BB(20G2) CDR-L3 | QQQHSYYT |
58 | 4-1BB(12B3) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR SEFRFYADFDYWGQGTTVTVSS |
59 | 4-1BB(12B3) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYHSYPQTFGQGTKVEIK |
60 | 4-1BB(25G7) VH | EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRQAPGKGLEWVS AISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAR DDPWPPFDYWGQGTLVTVSS |
61 | 4-1BB(25G7) VL | SSELTQDPAVSVALGQTVRITC QGDSLRSYYASWYQQKPGQAPVLVIY GKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYC NSLDRRGMWVFGGGTKLTV |
62 | 4-1BB(11D5) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR STLIYGYFDYWGQGTTVTVSS |
63 | 4-1BB(11D5) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQLNSYPQTFGQGTKVEIK |
64 | 4-1BB(9B11) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR SSGAYPGYFDYWGQGTTVTVSS |
65 | 4-1BB(9B11) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQVNSYPQTFGQGTKVEIK |
66 | 4-1BB(20G2) VH | QVQLVQSGAEVKKPGSSVKVSCKASGGTFS SYAISWVRQAPGQGLEWMG GIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAR SYYWESYPFDYWGQGTTVTVSS |
67 | 4-1BB(20G2) VL | DIQMTQSPSTLSASVGDRVTITC RASQSISSWLAWYQQKPGKAPKLLIY DASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQQHSYYTFGQGTKVEIK |
68 | FAP(28H1) CDR-H1 | SHAMS |
69 | FAP(4B9) CDR-H1 | SYAMS |
70 | FAP(28H1) CDR-H2 | AIWASGEQYYADSVKG |
71 | FAP(4B9) CDR-H2 | AIIGSGASTYYADSVKG |
72 | FAP(28H1) CDR-H3 | GWLGNFDY |
73 | FAP(4B9) CDR-H3 | GWFGGFNY |
74 | FAP(28H1) CDR-L1 | RASQSVSRSYLA |
75 | FAP(4B9) CDR-L1 | RASQSVTSSYLA |
76 | FAP(28H1) CDR-L2 | GASTRAT |
77 | FAP(4B9) CDR-L2 | VGSRRAT |
78 | FAP(28H1) CDR-L3 | QQGQVIPPT |
79 | FAP(4B9) CDR-L3 | QQGIMLPPT |
80 | FAP(28H1) VH | EVQLLESGGGLVQPGGSLRLSCAASGFTFS SHAMSWVRQAPGKGLEWVS AIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GWLGNFDYWGQGTLVTVSS |
81 | FAP(28H1) VL | EIVLTQSPGTLSLSPGERATLSC RASQSVSRSYLAWYQQKPGQAPRLLII GASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQGQVIPPTFGQGTKVEIK |
82 | FAP(4B9) VH | EVQLLESGGGLVQPGGSLRLSCAASGFTFS SYAMSWVRQAPGKGLEWVS AIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAK GWFGGFNYWGQGTLVTVSS |
83 | FAP(4B9) VL | EIVLTQSPGTLSLSPGERATLSC RASQSVTSSYLAWYQQKPGQAPRLLIN VGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC QQGIMLPPTFGQGTKVEIK |
84 | 人類(hu) FAP | UniProt編號Q12884 |
85 | hu FAP胞外域+聚-lys-標籤+his 6-標籤 | RPSRVHNSEENTMRALTLKDILNGTFSYKTFFPNWISGQEYLHQSADNNIVLYNIETGQSYTILSNRTMKSVNASNYGLSPDRQFVYLESDYSKLWRYSYTATYYIYDLSNGEFVRGNELPRPIQYLCWSPVGSKLAYVYQNNIYLKQRPGDPPFQITFNGRENKIFNGIPDWVYEEEMLATKYALWWSPNGKFLAYAEFNDTDIPVIAYSYYGDEQYPRTINIPYPKAGAKNPVVRIFIIDTTYPAYVGPQEVPVPAMIASSDYYFSWLTWVTDERVCLQWLKRVQNVSVLSICDFREDWQTWDCPKTQEHIEESRTGWAGGFFVSTPVFSYDAISYYKIFSDKDGYKHIHYIKDTVENAIQITSGKWEAINIFRVTQDSLFYSSNEFEEYPGRRNIYRISIGSYPPSKKCVTCHLRKERCQYYTASFSDYAKYYALVCYGPGIPISTLHDGRTDQEIKILEENKELENALKNIQLPKEEIKKLEVDEITLWYKMILPPQFDRSKKYPLLIQVYGGPCSQSVRSVFAVNWISYLASKEGMVIALVDGRGTAFQGDKLLYAVYRKLGVYEVEDQITAVRKFIEMGFIDEKRIAIWGWSYGGYVSSLALASGTGLFKCGIAVAPVSSWEYYASVYTERFMGLPTKDDNLEHYKNSTVMARAEYFRNVDYLLIHGTADDNVHFQNSAQIAKALVNAQVDFQAMWYSDQNHGLSGLSTNHLYTHMTHFLKQCFSLSDGKKKKKKGHHHHHH |
86 | 核苷酸序列 hu FAP胞外域+聚-lys-標籤+his 6-標籤 | CGCCCTTCAAGAGTTCATAACTCTGAAGAAAATACAATGAGAGCACTCACACTGAAGGATATTTTAAATG GAACATTTTCTTATAAAACATTTTTTCCAAACTGGATTTCAGGACAAGAATATCTTCATCAATCTGCAGATAACAATATAGTACTTTATAATATTGAAACAGGACAATCATATACCATTTTGAGTAATAGAACCATGAAAAGTGTGAATGCTTCAAATTACGGCTTATCACCTGATCGGCAATTTGTATATCTAGAAAGTGATTATTCAAAGCTTTGGAGATACTCTTACACAGCAACATATTACATCTATGACCTTAGCAATGGAGAATTTGTAAGAGGAAATGAGCTTCCTCGTCCAATTCAGTATTTATGCTGGTCGCCTGTTGGGAGTAAATTAGCATATGTCTATCAAAACAATATCTATTTGAAACAAAGACCAGGAGATCCACCTTTTCAAATAACATTTAATGGAAGAGAAAATAAAATATTTAATGGAATCCCAGACTGGGTTTATGAAGAGGAAATGCTTGCTACAAAATATGCTCTCTGGTGGTCTCCTAATGGAAAATTTTTGGCATATGCGGAATTTAATGATACGGATATACCAGTTATTGCCTATTCCTATTATGGCGATGAACAATATCCTAGAACAATAAATATTCCATACCCAAAGGCTGGAGCTAAGAATCCCGTTGTTCGGATATTTATTATCGATACCACTTACCCTGCGTATGTAGGTCCCCAGGAAGTGCCTGTTCCAGCAATGATAGCCTCAAGTGATTATTATTTCAGTTGGCTCACGTGGGTTACTGATGAACGAGTATGTTTGCAGTGGCTAAAAAGAGTCCAGAATGTTTCGGTCCTGTCTATATGTGACTTCAGGGAAGACTGGCAGACATGGGATTGTCCAAAGACCCAGGAGCATATAGAAGAAAGCAGAACTGGATGGGCTGGTGGATTCTTTGTTTCAACACCAGTTTTCAGCTATGATGCCATTTCGTACTACAAAATATTTAGTGACAAGGATGGCTACAAACATATTCACTATATCAAAGACACTGTGGAAAATGCTATTCAAATTACAAGTGGCAAGTGGGAGGCCATAAATATATTCAGAGTAACACAGGATTCACTGTTTTATTCTAGCAATGAATTTGAAGAATACCCTGGAAGAAGAAACATCTACAGAATTAGCATTGGAAGCTATCCTCCAAGCAAGAAGTGTGTTACTTGCCATCTAAGGAAAGAAAGGTGCCAATATTACACAGCAAGTTTCAGCGACTACGCCAAGTACTATGCACTTGTCTGCTACGGCCCAGGCATCCCCATTTCCACCCTTCATGATGGACGCACTGATCAAGAAATTAAAATCCTGGAAGAAAACAAGGAATTGGAAAATGCTTTGAAAAATATCCAGCTGCCTAAAGAGGAAATTAAGAAACTTGAAGTAGATGAAATTACTTTATGGTACAAGATGATTCTTCCTCCTCAATTTGACAGATCAAAGAAGTATCCCTTGCTAATTCAAGTGTATGGTGGTCCCTGCAGTCAGAGTGTAAGGTCTGTATTTGCTGTTAATTGGATATCTTATCTTGCAAGTAAGGAAGGGATGGTCATTGCCTTGGTGGATGGTCGAGGAACAGCTTTCCAAGGTGACAAACTCCTCTATGCAGTGTATCGAAAGCTGGGTGTTTATGAAGTTGAAGACCAGATTACAGCTGTCAGAAAATTCATAGAAATGGGTTTCATTGATGAAAAAAGAATAGCCATATGGGGCTGGTCCTATGGAGGATACGTTTCATCACTGGCCCTTGCATCTGGAACTGGTCTTTTCAAATGTGGTATAGCAGTGGCTCCAGTCTCCAGCTGGGAATATTACGCGTCTGTCTACACAGAGAGATTCATGGGTCTCCCAACAAAGGATGATAATCTTGAGCACTATAAGAATTCAACTGTGATGGCAAGAGCAGAATATTTCAGAAATGTAGACTATCTTCTCATCCACGGAACAGCAGATGATAATGTGCACTTTCAAAACTCAGCACAGATTGCTAAAGCTCTGGTTAATGCACAAGTGGATTTCCAGGCAATGTGGTACTCTGACCAGAACCACGGCTTATCCGGCCTGTCCACGAACCACTTATACACCCACATGACCCACTTCCTAAAGCAGTGTTTCTCTTTGTCAGACGGCAAAAAGAAAAAGAAAAAGGGCCACCACCATCACCATCAC |
87 | 小鼠FAP | UniProt編號P97321 |
88 | 鼠類FAP胞外域+聚-lys-標籤+his 6-標籤 | RPSRVYKPEGNTKRALTLKDILNGTFSYKTYFPNWISEQEYLHQSEDDNIVFYNIETRESYIILSNSTMKSVNATDYGLSPDRQFVYLESDYSKLWRYSYTATYYIYDLQNGEFVRGYELPRPIQYLCWSPVGSKLAYVYQNNIYLKQRPGDPPFQITYTGRENRIFNGIPDWVYEEEMLATKYALWWSPDGKFLAYVEFNDSDIPIIAYSYYGDGQYPRTINIPYPKAGAKNPVVRVFIVDTTYPHHVGPMEVPVPEMIASSDYYFSWLTWVSSERVCLQWLKRVQNVSVLSICDFREDWHAWECPKNQEHVEESRTGWAGGFFVSTPAFSQDATSYYKIFSDKDGYKHIHYIKDTVENAIQITSGKWEAIYIFRVTQDSLFYSSNEFEGYPGRRNIYRISIGNSPPSKKCVTCHLRKERCQYYTASFSYKAKYYALVCYGPGLPISTLHDGRTDQEIQVLEENKELENSLRNIQLPKVEIKKLKDGGLTFWYKMILPPQFDRSKKYPLLIQVYGGPCSQSVKSVFAVNWITYLASKEGIVIALVDGRGTAFQGDKFLHAVYRKLGVYEVEDQLTAVRKFIEMGFIDEERIAIWGWSYGGYVSSLALASGTGLFKCGIAVAPVSSWEYYASIYSERFMGLPTKDDNLEHYKNSTVMARAEYFRNVDYLLIHGTADDNVHFQNSAQIAKALVNAQVDFQAMWYSDQNHGILSGRSQNHLYTHMTHFLKQCFSLSDGKKKKKKGHHHHHH |
89 | 核苷酸序列 鼠類FAP胞外域+聚-lys-標籤+his 6-標籤 | CGTCCCTCAAGAGTTTACAAACCTGAAGGAAACACAAAGAGAGCTCTTACCTTGAAGGATATTTTAAATG GAACATTCTCATATAAAACATATTTTCCCAACTGGATTTCAGAACAAGAATATCTTCATCAATCTGAGGATGATAACATAGTATTTTATAATATTGAAACAAGAGAATCATATATCATTTTGAGTAATAGCACCATGAAAAGTGTGAATGCTACAGATTATGGTTTGTCACCTGATCGGCAATTTGTGTATCTAGAAAGTGATTATTCAAAGCTCTGGCGATATTCATACACAGCGACATACTACATCTACGACCTTCAGAATGGGGAATTTGTAAGAGG ATACGAGCTCCCTCGTCCAATTCAGTATCTATGCTGGTCGCCTGTTGGGAGTAAATTAGCATATGTATATCAAAACAATATTTATTTGAAACAAAGACCAGGAGATCCACCTTTTCAAATAACTTATACTGGAAGAGAAAATAGAATATTTAATGGAATACCAGACTGGGTTTATGAAGAGGAAATGCTTGCCACAAAATATGCTCTTTGGTGGTCTCCAGATGGAAAATTTTTGGCATATGTAGAATTTAATGATTCAGATATACCAATTATTGCCTATTCTTATTATGGTGATGGACAGTATCCTAGAACTATAAATATTCCATATCCAAAGGCTGGGGCTAAGAATCCGGTTGTTCGTGTTTTTATTGTTGACACCACCTACCCTCACCACGTGGGCCCAATGGAAGTGCCAGTTCCAGAAATGATAGCCTCAAGTGACTATTATTTCAGCTGGCTCACATGGGTGTCCAGTGAACGAGTATGCTTGCAGTGGCTAAAAAGAGTGCAGAATGTCTCAGTCCTGTCTATATGTGATTTCAGGGAAGACTGGCATGCATGGGAATGTCCAAAGAACCAGGAGCATGTAGAAGAAAGCAGAACAGGATGGGCTGGTGGATTCTTTGTTTCGACACCAGCTTTTAGCCAGGATGCCACTTCTTACTACAAAATATTTAGCGACAAGGATGGTTACAAACATATTCACTACATCAAAGACACTGTGGAAAATGCTATTCAAATTACAAGTGGCAAGTGGGAGGCCATATATATATTCCGCGTAACACAGGATTCACTGTTTTATTCTAGCAATGAATTTGAAGGTTACCCTGGAAGAAGAAACATCTACAGAATTAGCATTGGAAACTCTCCTCCGAGCAAGAAGTGTGTTACTTGCCATCTAAGGAAAGAAAGGTGCCAATATTACACAGCAAGTTTCAGCTACAAAGCCAAGTACTATGCACTCGTCTGCTATGGCCCTGGCCTCCCCATTTCCACCCTCCATGATGGCCGCACAGACCAAGAAATACAAGTATTAGAAGAAAACAAAGAACTGGAAAATTCTCTGAGAAATATCCAGCTGCCTAAAGTGGAGATTAAGAAGCTCAAAGACGGGGGACTGACTTTCTGGTACAAGATGATTCTGCCTCCTCAGTTTGACAGATCAAAGAAGTACCCTTTGCTAATTCAAGTGTATGGTGGTCCTTGTAGCCAGAGTGTTAAGTCTGTGTTTGCTGTTAATTGGATAACTTATCTCGCAAGTAAGGAGGGGATAGTCATTGCCCTGGTAGATGGTCGGGGCACTGCTTTCCAAGGTGACAAATTCCTGCATGCCGTGTATCGAAAACTGGGTGTATATGAAGTTGAGGACCAGCTCACAGCTGTCAGAAAATTCATAGAAATGGGTTTCATTGATGAAGAAAGAATAGCCATATGGGGCTGGTCCTACGGAGGTTATGTTTCATCCCTGGCCCTTGCATCTGGAACTGGTCTTTTCAAATGTGGCATAGCAGTGGCTCCAGTCTCCAGCTGGGAATATTACGCATCTATCTACTCAGAGAGATTCATGGGCCTCCCAACAAAGGACGACAATCTCGAACACTATAAAAATTCAACTGTGATGGCAAGAGCAGAATATTTCAGAAATGTAGACTATCTTCTCATCCACGGAACAGCAGATGATAATGTGCACTTTCAGAACTCAGCACAGATTGCTAAAGCTTTGGTTAATGCACAAGTGGATTTCCAGGCGATGTGGTACTCTGACCAGAACCATGGTATATTATCTGGGCGCTCCCAGAATCATTTATATACCCACATGACGCACTTCCTCAAGCAATGCTTTTCTTTATCAGACGGCAAAAAGAAAAAGAAAAAGGGCCACCACCATCACCATCAC |
90 | 獼猴FAP胞外域+聚-lys-標籤+his 6-標籤 | RPPRVHNSEENTMRALTLKDILNGTFSYKTFFPNWISGQEYLHQSADNNIVLYNIETGQSYTILSNRTMKSVNASNYGLSPDRQFVYLESDYSKLWRYSYTATYYIYDLSNGEFVRGNELPRPIQYLCWSPVGSKLAYVYQNNIYLKQRPGDPPFQITFNGRENKIFNGIPDWVYEEEMLATKYALWWSPNGKFLAYAEFNDTDIPVIAYSYYGDEQYPRTINIPYPKAGAKNPFVRIFIIDTTYPAYVGPQEVPVPAMIASSDYYFSWLTWVTDERVCLQWLKRVQNVSVLSICDFREDWQTWDCPKTQEHIEESRTGWAGGFFVSTPVFSYDAISYYKIFSDKDGYKHIHYIKDTVENAIQITSGKWEAINIFRVTQDSLFYSSNEFEDYPGRRNIYRISIGSYPPSKKCVTCHLRKERCQYYTASFSDYAKYYALVCYGPGIPISTLHDGRTDQEIKILEENKELENALKNIQLPKEEIKKLEVDEITLWYKMILPPQFDRSKKYPLLIQVYGGPCSQSVRSVFAVNWISYLASKEGMVIALVDGRGTAFQGDKLLYAVYRKLGVYEVEDQITAVRKFIEMGFIDEKRIAIWGWSYGGYVSSLALASGTGLFKCGIAVAPVSSWEYYASVYTERFMGLPTKDDNLEHYKNSTVMARAEYFRNVDYLLIHGTADDNVHFQNSAQIAKALVNAQVDFQAMWYSDQNHGLSGLSTNHLYTHMTHFLKQCFSLSDGKKKKKKGHHHHHH |
91 | 核苷酸序列 獼猴FAP胞外域+聚-lys-標籤+his 6-標籤 | CGCCCTCCAAGAGTTCATAACTCTGAAGAAAATACAATGAGAGCACTCACACTGAAGGATATTTTAAATG GGACATTTTCTTATAAAACATTTTTTCCAAACTGGATTTCAGGACAAGAATATCTTCATCAATCTGCAGATAACAATATAGTACTTTATAATATTGAAACAGGACAATCATATACCATTTTGAGTAACAGAACCATGAAAAGTGTGAATGCTTCAAATTATGGCTTATCACCTGATCGGCAATTTGTATATCTAGAAAGTGATTATTCAAAGCTTTGGAGATACTCTTACACAGCAACATATTACATCTATGACCTTAGCAATGGAGAATTTGTAAGAGGAAATGAGCTTCCTCGTCCAATTCAGTATTTATGCTGGTCGCCTGTTGGGAGTAAATTAGCATATGTCTATCAAAACAATATCTATTTGAAACAAAGACCAGGAGATCCACCTTTTCAAATAACATTTAATGGAAGAGAAAATAAAATATTTAATGGAATCCCAGACTGGGTTTATGAAGAGGAAATGCTTGCTACAAAATATGCTCTCTGGTGGTCTCCTAATGGAAAATTTTTGGCATATGCGGAATTTAATGATACAGATATACCAGTTATTGCCTATTCCTATTATGGCGATGAACAATATCCCAGAACAATAAATATTCCATACCCAAAGGCCGGAGCTAAGAATCCTTTTGTTCGGATATTTATTATCGATACCACTTACCCTGCGTATGTAGGTCCCCAGGAAGTGCCTGTTCCAGCAATGATAGCCTCAAGTGATTATTATTTCAGTTGGCTCACGTGGGTTACTGATGAACGAGTATGTTTGCAGTGGCTAAAAAGAGTCCAGAATGTTTCGGTCTTGTCTATATGTGATTTCAGGGAAGACTGGCAGACATGGGATTGTCCAAAGACCCAGGAGCATATAGAAGAAAGCAGAACTGGATGGGCTGGTGGATTCTTTGTTTCAACACCAGTTTTCAGCTATGATGCCATTTCATACTACAAAATATTTAGTGACAAGGATGGCTACAAACATATTCACTATATCAAAGACACTGTGGAAAATGCTATTCAAATTACAAGTGGCAAGTGGGAGGCCATAAATATATTCAGAGTAACACAGGATTCACTGTTTTATTCTAGCAATGAATTTGAAGATTACCCTGGAAGAAGAAACATCTACAGAATTAGCATTGGAAGCTATCCTCCAAGCAAGAAGTGTGTTACTTGCCATCTAAGGAAAGAAAGGTGCCAATATTACACAGCAAGTTTCAGCGACTACGCCAAGTACTATGCACTTGTCTGCTATGGCCCAGGCATCCCCATTTCCACCCTTCATGACGGACGCACTGATCAAGAAATTAAAATCCTGGAAGAAAACAAGGAATTGGAAAATGCTTTGAAAAATATCCAGCTGCCTAAAGAGGAAATTAAGAAACTTGAAGTAGATGAAATTACTTTATGGTACAAGATGATTCTTCCTCCTCAATTTGACAGATCAAAGAAGTATCCCTTGCTAATTCAAGTGTATGGTGGTCCCTGCAGTCAGAGTGTAAGGTCTGTATTTGCTGTTAATTGGATATCTTATCTTGCAAGTAAGGAAGGGATGGTCATTGCCTTGGTGGATGGTCGGGGAACAGCTTTCCAAGGTGACAAACTCCTGTATGCAGTGTATCGAAAGCTGGGTGTTTATGAAGTTGAAGACCAGATTACAGCTGTCAGAAAATTCATAGAAATGGGTTTCATTGATGAAAAAAGAATAGCCATATGGGGCTGGTCCTATGGAGGATATGTTTCATCACTGGCCCTTGCATCTGGAACTGGTCTTTTCAAATGTGGGATAGCAGTGGCTCCAGTCTCCAGCTGGGAATATTACGCGTCTGTCTACACAGAGAGATTCATGGGTCTCCCAACAAAGGATGATAATCTTGAGCACTATAAGAATTCAACTGTGATGGCAAGAGCAGAATATTTCAGAAATGTAGACTATCTTCTCATCCACGGAACAGCAGATGATAATGTGCACTTTCAAAACTCAGCACAGATTGCTAAAGCTCTGGTTAATGCACAAGTGGATTTCCAGGCAATGTGGTACTCTGACCAGAACCACGGCTTATCCGGCCTGTCCACGAACCACTTATACACCCACATGACCCACTTCCTAAAGCAGTGTTTCTCTTTGTCAGACGGCAAAAAGAAAAAGAAAAAGGGCCACCACCATCACCATCAC |
92 | 人類CEA | UniProt編號P06731 |
93 | 人類MCSP | UniProt編號Q6UVK1 |
94 | 人類EGFR | UniProt編號P00533 |
95 | 人類CD19 | UniProt編號P15391 |
96 | 人類CD20 | UniProt編號P11836 |
97 | 人類CD33 | UniProt編號P20138 |
98 | 人類OX40 | UniProt編號P43489 |
99 | 人類4-1BB | UniProt編號Q07011 |
100 | 人類CD27 | UniProt編號P26842 |
101 | 人類HVEM | UniProt編號Q92956 |
102 | 人類CD30 | UniProt編號P28908 |
103 | 人類GITR | UniProt編號Q9Y5U5 |
104 | 鼠類OX40 | UniProt編號P47741 |
105 | 鼠類4-1BB | UniProt編號P20334 |
106 | 獼猴4-1BB | UniProt編號F6W5G6 |
107 | 肽連接子(G4S) | GGGGS |
108 | 肽連接子(G4S) 2 | GGGGSGGGGS |
109 | 肽連接子(SG4) 2 | SGGGGSGGGG |
110 | 肽連接子G4(SG4) 2 | GGGGSGGGGSGGGG |
111 | 肽連接子 | GSPGSSSSGS |
112 | 肽連接子(G4S) 3 | GGGGSGGGGSGGGGS |
113 | 肽連接子(G4S) 4 | GGGGSGGGGSGGGGSGGGGS |
114 | 肽連接子 | GSGSGSGS |
115 | 肽連接子 | GSGSGNGS |
116 | 肽連接子 | GGSGSGSG |
117 | 肽連接子 | GGSGSG |
118 | 肽連接子 | GGSG |
119 | 肽連接子 | GGSGNGSG |
120 | 肽連接子 | GGNGSGSG |
121 | 肽連接子 | GGNGSG |
122 | 獼猴Ox40 ECD | aa 29-214 |
123 | 鼠類OX40 ECD | aa 10-211 |
124 | 核苷酸序列 Fc臼鏈 | 參看表2 |
125 | 核苷酸序列 人類OX40抗原Fc杵鏈 | 參看表2 |
126 | 核苷酸序列 獼猴OX40抗原Fc杵鏈 | 參看表2 |
127 | 核苷酸序列 鼠類OX40抗原Fc杵鏈 | 參看表2 |
128 | Fc臼鏈 | 參看表2 |
129 | 人類OX40抗原Fc杵鏈 | 參看表2 |
130 | 獼猴OX40抗原Fc杵鏈 | 參看表2 |
131 | 鼠類OX40抗原Fc杵鏈 | 參看表2 |
132 | 文庫DP88-4之核苷酸序列 | 參看表3 |
133 | Fab輕鏈Vk1_5之核苷酸序列 | 參看表4 |
134 | Fab輕鏈Vk1_5 | 參看表4 |
135 | Fab 重鏈VH1_69之核苷酸序列 | 參看表4 |
136 | Fab重鏈VH1_69 | 參看表4 |
137 | LMB3 | 參看表5 |
138 | Vk1_5_L3r_S | 參看表5 |
139 | Vk1_5_L3r_SY | 參看表5 |
140 | Vk1_5_L3r_SPY | 參看表5 |
141 | RJH31 | 參看表5 |
142 | RJH32 | 參看表5 |
143 | DP88-v4-4 | 參看表5 |
144 | DP88-v4-6 | 參看表5 |
145 | DP88-v4-8 | 參看表5 |
146 | fdseqlong | 參看表5 |
147 | (Vk3_20/VH3_23)模板 | 參看表6 |
148 | Fab輕鏈Vk3_20之核苷酸序列 | 參看表7 |
149 | Fab輕鏈Vk3_20 | 參看表7 |
150 | Fab重鏈VH3_23之核苷酸序列 | 參看表7 |
151 | Fab重鏈VH3_23 (DP47) | 參看表7 |
152 | MS64 | 參看表8 |
153 | DP47CDR3_ba (mod.) | 參看表8 |
154 | DP47-v4-4 | 參看表8 |
155 | DP47-v4-6 | 參看表8 |
156 | DP47-v4-8 | 參看表8 |
157 | fdseqlong | 參看表8 |
158 | Vl3_19/VH3_23文庫模板 | 參看表9 |
159 | Fab輕鏈Vl3_19之核苷酸序列 | 參看表10 |
160 | Fab輕鏈Vl3_19 | 參看表10 |
161 | LMB3 | 參看表11 |
162 | Vl_3_19_L3r_V | 參看表11 |
163 | Vl_3_19_L3r_HV | 參看表11 |
164 | Vl_3_19_L3r_HLV | 參看表11 |
165 | RJH80 | 參看表11 |
166 | 核苷酸序列 OX40(8H9) VL | 參看表12 |
167 | 核苷酸序列 OX40(8H9) VH | 參看表12 |
168 | 核苷酸序列 OX40(49B4) VL | 參看表12 |
169 | 核苷酸序列 OX40(49B4) VH | 參看表12 |
170 | 核苷酸序列 OX40(1G4) VL | 參看表12 |
171 | 核苷酸序列 OX40(1G4) VH | 參看表12 |
172 | 核苷酸序列 OX40(20B7) VL | 參看表12 |
173 | 核苷酸序列 OX40(20B7) VH | 參看表12 |
174 | 核苷酸序列 OX40(CLC-563) VL | 參看表12 |
175 | 核苷酸序列 OX40(CLC-563) VH | 參看表12 |
176 | 核苷酸序列 OX40(CLC-564) VL | 參看表12 |
177 | 核苷酸序列 OX40(CLC-564) VH | 參看表12 |
178 | 核苷酸序列 OX40(17A9) VL | 參看表12 |
179 | 核苷酸序列 OX40(17A9) VH | 參看表12 |
180 | 8H9 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
181 | 8H9 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
182 | 8H9 P329GLALA IgG1 (輕鏈) | 參看表13 |
183 | 8H9 P329GLALA IgG1 (重鏈) | 參看表13 |
184 | 49B4 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
185 | 49B4 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
186 | 49B4 P329GLALA IgG1 (輕鏈) | 參看表13 |
187 | 49B4 P329GLALA IgG1 (重鏈) | 參看表13 |
188 | 1G4 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
189 | 1G4 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
190 | 1G4 P329GLALA IgG1 (輕鏈) | 參看表13 |
191 | 1G4 P329GLALA IgG1 (重鏈) | 參看表13 |
192 | 20B 7P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
193 | 20B7 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
194 | 20B7 P329GLALA IgG1 (輕鏈) | 參看表13 |
195 | 20B7 P329GLALA IgG1 (重鏈) | 參看表13 |
196 | CLC-563 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
197 | CLC-563 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
198 | CLC-563 P329GLALA IgG1 (輕鏈) | 參看表13 |
199 | CLC-563 P329GLALA IgG1 (重鏈) | 參看表13 |
200 | CLC-564 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
201 | CLC-564 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
202 | CLC-564 P329GLALA IgG1 (輕鏈) | 參看表13 |
203 | CLC-564 P329GLALA IgG1 (重鏈) | 參看表13 |
204 | 17A9 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表13 |
205 | 17A9 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表13 |
206 | 17A9 P329GLALA IgG1 (輕鏈) | 參看表13 |
207 | 17A9 P329GLALA IgG1 (重鏈) | 參看表13 |
208 | 人類OX40 His | 核苷酸序列 |
209 | 人類OX40 His | 參看表15 |
210 | 鼠類OX40 His | 核苷酸序列 |
211 | 鼠類OX40 His | 參看表15 |
212 | 嵌合人類OX40抗原Fc之核苷酸序列 | 參看表21 |
213 | 嵌合人類OX40抗原Fc | 參看表21 |
214 | (8B9) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表25 |
215 | VLCH1-輕鏈 2 (28H1) | 核苷酸序列,參看表25 |
216 | (8B9) VHCH1-重鏈-(28H1) VHCL | 參看表25 |
217 | VLCH1-輕鏈2 (28H1) | 參看表25 |
218 | (49B4) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表25 |
219 | (49B4) VHCH1-重鏈-(28H1) VHCL | 參看表25 |
220 | (1G4) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表25 |
221 | (1G4) VHCH1-重鏈-(28H1) VHCL | 參看表25 |
222 | (20B7) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表25 |
223 | (20B7) VHCH1-重鏈-(28H1) VHCL | 參看表25 |
224 | (CLC-563) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表25 |
225 | (CLC-563) VHCH1-重鏈-(28H1) VHCL | 參看表25 |
226 | (CLC-564) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表25 |
227 | (CLC-564) VHCH1-重鏈-(28H1) VHCL | 參看表25 |
228 | (28H1) VHCL-重鏈臼 | 核苷酸序列,參看表27 |
229 | (28H1) VHCL-重鏈臼 | 參看表27 |
230 | (49B4) VHCH1-重鏈杵 | 核苷酸序列,參看表27 |
231 | (49B4) VHCH1-重鏈杵 | 參看表27 |
232 | (1G4) VHCH1-重鏈杵 | 核苷酸序列,參看表27 |
233 | (1G4) VHCH1-重鏈杵 | 參看表27 |
234 | (20B7) VHCH1-重鏈杵 | 核苷酸序列,參看表27 |
235 | (20B7) VHCH1-重鏈杵 | 參看表27 |
236 | (CLC-563) VHCH1-重鏈杵 | 核苷酸序列,參看表27 |
237 | (CLC-563) VHCH1-重鏈杵 | 參看表27 |
238 | (CLC-564) VHCH1-重鏈杵 | 核苷酸序列,參看表27 |
239 | (CLC-564) VHCH1-重鏈杵 | 參看表27 |
240 | 獼猴4-1BB ECD | aa 24-186 |
241 | 鼠類4-1BB ECD | P20334, aa 24-187 |
242 | 人類4-1BB抗原Fc杵鏈 | 核苷酸序列,參看表37 |
243 | 獼猴4-1BB抗原Fc杵鏈 | 核苷酸序列,參看表37 |
244 | 鼠類4-1BB抗原Fc杵鏈 | 核苷酸序列,參看表37 |
245 | 人類4-1BB抗原Fc杵鏈 | 參看表37 |
246 | 獼猴4-1BB抗原 Fc杵鏈 | 參看表37 |
247 | 鼠類4-1BB抗原Fc杵鏈 | 參看表37 |
248 | 引子MS63 | 參看表43 |
249 | 核苷酸序列 4-1BB(12B3) VL | 參看表44 |
250 | 核苷酸序列 4-1BB(12B3) VH | 參看表44 |
251 | 核苷酸序列 4-1BB(25G7) VL | 參看表44 |
252 | 核苷酸序列 4-1BB(25G7) VH | 參看表44 |
253 | 核苷酸序列 4-1BB(11D5) VL | 參看表44 |
254 | 核苷酸序列 4-1BB(11D5) VH | 參看表44 |
255 | 核苷酸序列 4-1BB(9B11) VL | 參看表44 |
256 | 核苷酸序列 4-1BB(9B11) VH | 參看表44 |
257 | 核苷酸序列 4-1BB(20G2) VL | 參看表44 |
258 | 核苷酸序列 4-1BB(20G2) VH | 參看表44 |
259 | 12B3 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表45 |
260 | 12B3 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表45 |
261 | 12B3 P329GLALA IgG1 (輕鏈) | 參看表45 |
262 | 12B3 P329GLALA IgG1 (重鏈) | 參看表45 |
263 | 25G7 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表45 |
264 | 25G7 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表45 |
265 | 25G7 P329GLALA IgG1 (輕鏈) | 參看表45 |
266 | 25G7 P329GLALA IgG1 (重鏈) | 參看表45 |
267 | 11D5 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表45 |
268 | 11D5 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表45 |
269 | 11D5 P329GLALA IgG1 (輕鏈) | 參看表45 |
270 | 11D5 P329GLALA IgG1 (重鏈) | 參看表45 |
271 | 9B11 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表45 |
272 | 9B11 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表45 |
273 | 9B11 P329GLALA IgG1 (輕鏈) | 參看表45 |
274 | 9B11 P329GLALA IgG1 (重鏈) | 參看表45 |
275 | 20G2 P329GLALA IgG1 (輕鏈) | 核苷酸序列,參看表45 |
276 | 20G2 P329GLALA IgG1 (重鏈) | 核苷酸序列,參看表45 |
277 | 20G2 P329GLALA IgG1 (輕鏈) | 參看表45 |
278 | 20G2 P329GLALA IgG1 (重鏈) | 參看表45 |
279 | mu4-1BB D1 / hu4-1BB D2 Fc杵 | 參看表56 |
280 | hu4-1BB D1 / mu4-1BB D2 Fc杵 | 參看表56 |
281 | hu4-1BB D1 Fc杵 | 參看表56 |
282 | 鼠類4-1BB結構域D1 (N端) | 參看表57 |
283 | 人類4-1BB結構域D2 (C端) | 參看表57 |
284 | 人類4-1BB結構域D1 (N端) | 參看表57 |
285 | 鼠類4-1BB結構域D2 (C端) | 參看表52 |
286 | (12B3) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表60 |
287 | (12B3) VHCH1-重鏈-(28H1) VHCL | 參看表60 |
288 | (25G7) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表60 |
289 | (25G7) VHCH1-重鏈-(28H1) VHCL | 參看表60 |
290 | (11D5) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表60 |
291 | (11D5) VHCH1-重鏈-(28H1) VHCL | 參看表60 |
292 | (9B11) VHCH1-重鏈-(28H1) VHCL | 核苷酸序列,參看表60 |
293 | (9B11) VHCH1-重鏈-(28H1) VHCL | 參看表60 |
294 | (12B3) VHCH1-重鏈杵 | 核苷酸序列,參看表65 |
295 | (12B3) VHCH1-重鏈杵 | 參看表65 |
296 | (25G7) VHCH1-重鏈杵 | 核苷酸序列,參看表65 |
297 | (25G7) VHCH1-重鏈杵 | 參看表65 |
298 | (11D5) VHCH1-重鏈杵 | 核苷酸序列,參看表65 |
299 | (11D5) VHCH1-重鏈杵 | 參看表65 |
300 | (9B11) VHCH1-重鏈杵 | 核苷酸序列,參看表65 |
301 | (9B11) VHCH1-重鏈杵 | 參看表65 |
302 | (8H9) VHCH1-重鏈杵 | 核苷酸序列,參看表27 |
303 | (8H9) VHCH1-重鏈杵 | 參看表27 |
304 | (49B4) VHCH1 Fc杵VH (4B9) (重鏈1之核苷酸序列) | 參看表30 |
305 | (49B4) VHCH1 Fc臼VL (4B9) (重鏈2之核苷酸序列) | 參看表30 |
306 | (49B4) VHCH1 Fc杵VH (4B9) (重鏈1) | 參看表30 |
307 | (49B4) VHCH1 Fc臼VL (4B9) (重鏈2) | 參看表30 |
308 | (49B4) VHCH1 Fc杵VH (28H1) (核苷酸序列,重鏈1) | 參看表30 |
309 | (49B4) VHCH1 Fc臼VL (28H1) (核苷酸序列,重鏈2) | 參看表30 |
310 | (49B4) VHCH1 Fc杵VH (28H1) (重鏈1) | 參看表30 |
311 | (49B4) VHCH1 Fc臼VL (28H1) (重鏈2) | 參看表30 |
312 | (49B4) VHCH1 Fc杵VH (DP47) (核苷酸序列,重鏈1) | 參看表30 |
313 | (49B4) VHCH1 Fc臼VL (DP47) (核苷酸序列,重鏈2) | 參看表30 |
314 | (49B4) VHCH1 Fc杵VH (DP47) (重鏈1) | 參看表30 |
315 | (49B4) VHCH1 Fc臼VL (DP47) (重鏈2) | 參看表30 |
316 | (12B3) VHCH1 Fc杵VH (4B9) (HC 1之核苷酸序列) | 參看表70 |
317 | (12B3) VHCH1 Fc臼VL (4B9) (HC2之核苷酸序列) | 參看表70 |
318 | (12B3) VHCH1 Fc杵VH (4B9) (重鏈1) | 參看表70 |
319 | (12B3) VHCH1 Fc臼VL (4B9) (重鏈2) | 參看表70 |
320 | (25G7) VHCH1 Fc杵VH (4B9) (核苷酸序列,重鏈1) | 參看表70 |
321 | (25G7) VHCH1 Fc臼VL (4B9) (核苷酸序列,重鏈2) | 參看表70 |
322 | (25G7) VHCH1 Fc杵VH (4B9) (重鏈1) | 參看表70 |
323 | (25G7) VHCH1 Fc臼VL (4B9) (重鏈2) | 參看表70 |
324 | (11D5) VHCH1 Fc杵VH (4B9) (核苷酸序列,重鏈1) | 參看表70 |
325 | (11D5) VHCH1 Fc臼VL (4B9) (核苷酸序列,重鏈2) | 參看表70 |
326 | (11D5) VHCH1 Fc杵VH (4B9) (重鏈1) | 參看表70 |
327 | (11D5) VHCH1 Fc臼VL (4B9) (重鏈2) | 參看表70 |
328 | (9B11) VHCH1 Fc杵VH (4B9) (核苷酸序列,重鏈1) | 參看表70 |
329 | (9B11) VHCH1 Fc臼VL (4B9) (核苷酸序列,重鏈2) | 參看表70 |
330 | (9B11) VHCH1 Fc杵VH (4B9) (重鏈1) | 參看表70 |
331 | (9B11) VHCH1 Fc臼VL (4B9) (重鏈2) | 參看表70 |
332 | (12B3) VHCH1 Fc杵VL (4B9) (HC 1之核苷酸序列) | 參看表71 |
333 | (12B3) VHCH1 Fc臼VH (4B9) (HC2之核苷酸序列) | 參看表71 |
334 | (12B3) VHCH1 Fc杵VL (4B9) (重鏈1) | 參看表71 |
335 | (12B3) VHCH1 Fc臼VH (4B9) (重鏈2) | 參看表71 |
336 | (25G7) VHCH1 Fc杵VL (4B9) (核苷酸序列,重鏈1) | 參看表71 |
337 | (25G7) VHCH1 Fc臼VH (4B9) (核苷酸序列,重鏈2) | 參看表71 |
338 | (25G7) VHCH1 Fc杵VL (4B9) (重鏈1) | 參看表71 |
339 | (25G7) VHCH1 Fc臼VH (4B9) (重鏈2) | 參看表71 |
340 | (11D5) VHCH1 Fc杵VL (4B9) (核苷酸序列,重鏈1) | 參看表71 |
341 | (11D5) VHCH1 Fc臼VH (4B9) (核苷酸序列,重鏈2) | 參看表71 |
342 | (11D5) VHCH1 Fc杵VL (4B9) (重鏈1) | 參看表71 |
343 | (11D5) VHCH1 Fc臼VH (4B9) (重鏈2) | 參看表71 |
344 | (9B11) VHCH1 Fc杵VL (4B9) (核苷酸序列,重鏈1) | 參看表71 |
345 | (9B11) VHCH1 Fc臼VH (4B9) (核苷酸序列,重鏈2) | 參看表71 |
346 | (9B11) VHCH1 Fc杵VL (4B9) (重鏈1) | 參看表71 |
347 | (9B11) VHCH1 Fc臼VH (4B9) (重鏈2) | 參看表71 |
SEQ ID NO: | 構築體 | 來源 | ECD |
1 | 人類OX40 ECD | 根據P43489合成 | aa 29-214 |
122 | 獼猴OX40 ECD | 自獼猴血液分離 | aa 29-214 |
123 | 鼠類OX40 ECD | 根據P47741合成 | aa 10-211 |
SEQ ID NO: | 抗原 | 序列 |
124 | 核苷酸序列 Fc臼鏈 | GACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
125 | 核苷酸序列 人類OX40抗原Fc杵鏈 | CTGCACTGCGTGGGCGACACCTACCCCAGCAACGACCGGTGCTGCCACGAGTGCAGACCCGGCAACGGCATGGTGTCCCGGTGCAGCCGGTCCCAGAACACCGTGTGCAGACCTTGCGGCCCTGGCTTCTACAACGACGTGGTGTCCAGCAAGCCCTGCAAGCCTTGTACCTGGTGCAACCTGCGGAGCGGCAGCGAGCGGAAGCAGCTGTGTACCGCCACCCAGGATACCGTGTGCCGGTGTAGAGCCGGCACCCAGCCCCTGGACAGCTACAAACCCGGCGTGGACTGCGCCCCTTGCCCTCCTGGCCACTTCAGCCCTGGCGACAACCAGGCCTGCAAGCCTTGGACCAACTGCACCCTGGCCGGCAAGCACACCCTGCAGCCCGCCAGCAATAGCAGCGACGCCATCTGCGAGGACCGGGATCCTCCTGCCACCCAGCCTCAGGAAACCCAGGGCCCTCCCGCCAGACCCATCACCGTGCAGCCTACAGAGGCCTGGCCCAGAACCAGCCAGGGGCCTAGCACCAGACCCGTGGAAGTGCCTGGCGGCAGAGCCGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGAGGCCTGAACGACATCTTCGAGGCCCAGAAGATTGAATGGCACGAG |
126 | 核苷酸序列 獼猴OX40抗原Fc杵鏈 | CTCCACTGTGTCGGGGACACCTACCCCAGCAACGACCGGTGCTGTCAGGAGTGCAGGCCAGGCAACGGGATGGTGAGCCGCTGCAACCGCTCCCAGAACACGGTGTGCCGTCCGTGCGGGCCCGGCTTCTACAACGACGTGGTCAGCGCCAAGCCCTGCAAGGCCTGCACATGGTGCAACCTCAGAAGTGGGAGTGAGCGGAAACAGCCGTGCACGGCCACACAGGACACAGTCTGCCGCTGCCGGGCGGGCACCCAGCCCCTGGACAGCTACAAGCCTGGAGTTGACTGTGCCCCCTGCCCTCCAGGGCACTTCTCCCCGGGCGACAACCAGGCCTGCAAGCCCTGGACCAACTGCACCTTGGCCGGGAAGCACACCCTGCAGCCAGCCAGCAATAGCTCGGACGCCATCTGTGAGGACAGGGACCCCCCACCCACACAGCCCCAGGAGACCCAGGGCCCCCCGGCCAGGCCCACCACTGTCCAGCCCACTGAAGCCTGGCCCAGAACCTCACAGAGACCCTCCACCCGGCCCGTGGAGGTCCCCAGGGGCCCTGCGGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGAGGCCTGAACGACATCTTCGAGGCCCAGAAGATTGAATGGCACGAG |
127 | 核苷酸序列 鼠類OX40抗原Fc杵鏈 | GTGACCGCCAGACGGCTGAACTGCGTGAAGCACACCTACCCCAGCGGCCACAAGTGCTGCAGAGAGTGCCAGCCCGGCCACGGCATGGTGTCCAGATGCGACCACACACGGGACACCCTGTGCCACCCTTGCGAGACAGGCTTCTACAACGAGGCCGTGAACTACGATACCTGCAAGCAGTGCACCCAGTGCAACCACAGAAGCGGCAGCGAGCTGAAGCAGAACTGCACCCCCACCCAGGATACCGTGTGCAGATGCAGACCCGGCACCCAGCCCAGACAGGACAGCGGCTACAAGCTGGGCGTGGACTGCGTGCCCTGCCCTCCTGGCCACTTCAGCCCCGGCAACAACCAGGCCTGCAAGCCCTGGACCAACTGCACCCTGAGCGGCAAGCAGACCAGACACCCCGCCAGCGACAGCCTGGATGCCGTGTGCGAGGACAGAAGCCTGCTGGCCACCCTGCTGTGGGAGACACAGCGGCCCACCTTCAGACCCACCACCGTGCAGAGCACCACCGTGTGGCCCAGAACCAGCGAGCTGCCCAGTCCTCCTACCCTCGTGACACCTGAGGGCCCCGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGAGGCCTGAACGACATCTTCGAGGCCCAGAAGATTGAATGGCACGAG |
128 | Fc臼鏈 | DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
129 | 人類OX40抗原Fc杵鏈 | LHCVGDTYPSNDRCCHECRPGNGMVSRCSRSQNTVCRPCGPGFYNDVVSSKPCKPCTWCNLRSGSERKQLCTATQDTVCRCRAGTQPLDSYKPGVDCAPCPPGHFSPGDNQACKPWTNCTLAGKHTLQPASNSSDAICEDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
130 | 獼猴OX40抗原Fc杵鏈 | LHCVGDTYPSNDRCCQECRPGNGMVSRCNRSQNTVCRPCGPGFYNDVVSAKPCKACTWCNLRSGSERKQPCTATQDTVCRCRAGTQPLDSYKPGVDCAPCPPGHFSPGDNQACKPWTNCTLAGKHTLQPASNSSDAICEDRDPPPTQPQETQGPPARPTTVQPTEAWPRTSQRPSTRPVEVPRGPAVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
131 | 鼠類OX40抗原Fc杵鏈 | VTARRLNCVKHTYPSGHKCCRECQPGHGMVSRCDHTRDTLCHPCETGFYNEAVNYDTCKQCTQCNHRSGSELKQNCTPTQDTVCRCRPGTQPRQDSGYKLGVDCVPCPPGHFSPGNNQACKPWTNCTLSGKQTRHPASDSLDAVCEDRSLLATLLWETQRPTFRPTTVQSTTVWPRTSELPSPPTLVTPEGPVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
SEQ ID NO: | 描述 | 序列 |
132 | pRJH33文庫模板DP88-4文庫之核苷酸序列;完全Fab編碼區,其輕鏈包含PelB前導序列 + Vk1_5 κ V結構域 + CL恆定域且包括標籤之重鏈包含PelB + VH1_69 V結構域 + CH1恆定域 | TGAAATACCTATTGCCTACGGCAGCCGCTGGATTGTTATTACTCGCGGCCCAGCCGGCCATGGCCGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTCTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCATAATAAGGCGCGCCAATTCTATTTCAAGGAGACAGTCATATGAAATACCTGCTGCCGACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCCAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGACTATCCCCAGGCGGTTACTATGTTATGGATGCCTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAAGCACTAGTGCCCATCACCATCACCATCACGCCGCGGCA |
SEQ ID NO: | 描述 | 序列 |
133 | Fab輕鏈Vk1_5之核苷酸序列 | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTCTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCA |
134 | Fab輕鏈Vk1_5 | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGAAEQKLISEEDLNGAADYKDDDDKGAA |
135 | Fab重鏈VH1_69之核苷酸序列 | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGACTATCCCCAGGCGGTTACTATGTTATGGATGCCTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAAGCACTAGTGCCCATCACCATCACCATCACGCCGCGGCA |
136 | Fab重鏈VH1_69 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARLSPGGYYVMDAWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDAAASTSAHHHHHHAAA |
SEQ ID NO: | 引子名稱 | 引子序列5 ' - 3 ' |
137 | LMB3 | CAGGAAACAGCTATGACCATGATTAC |
138 | Vk1_5_L3r_S | CTCGACTTTGGTGCCCTGGCCAAACGT S BA A TA C GA A TT A TA CTGTTGGCAGTAATAAGTTGCAAAATCAT 帶下劃線的:60%初始鹼基及40%隨機分組為M。 加粗及斜體:60%初始鹼基及40%隨機分組為N |
139 | Vk1_5_L3r_SY | CTCGACTTTGGTGCCCTGGCCAAACGT M HR S GR A TA C GA A TT A TA CTGTTGGCAGTAATAAGTTGCAAAATCAT 帶下劃線的:60%初始鹼基及40%隨機分組為M。 加粗及斜體:60%初始鹼基及40%隨機分組為N |
140 | Vk1_5_L3r_SPY | CTCGACTTTGGTGCCCTGGCCAAACGT M HH M SS S GR A TA C GA A TT A TA CTGTTGGCAGTAATAAGTTGCAAAATCAT 帶下劃線的:60%初始鹼基及40%隨機分組為M。 加粗及斜體:60%初始鹼基及40%隨機分組為N |
141 | RJH31 | ACGTTTGGCCAGGGCACCAAAGTCGAG |
142 | RJH32 | TCTCGCACAGTAATACACGGCGGTGTCC |
143 | DP88-v4-4 | GGACACCGCCGTGTATTACTGTGCGAGA-1-2-2-3-4-GAC-TAC-TGGGGCCAAGGGACCACCGTGACCGTCTCC 1: G/D = 20%, E/V/S = 10%, A/P/R/L/T/Y=5%; 2: G/Y/S=15%, A/D/T/R/P/L/V/N/W/F/I/E = 4,6%; 3: G/A/Y = 20%, P/W/S/D/T = 8%; 4: F = 46%, L/M = 15%, G/I/Y = 8%. |
144 | DP88-v4-6 | GGACACCGCCGTGTATTACTGTGCGAGA-1-2-2-2-2-3-4-GAC-TAC-TGGGGCCAAGGGACCACCGTGACCGTCTCC 1: G/D = 20%, E/V/S = 10%, A/P/R/L/T/Y=5%; 2: G/Y/S=15%, A/D/T/R/P/L/V/N/W/F/I/E = 4,6%; 3: G/A/Y = 20%, P/W/S/D/T = 8%; 4: F = 46%, L/M = 15%, G/I/Y = 8%. |
145 | DP88-v4-8 | GGACACCGCCGTGTATTACTGTGCGAGA-1-2-2-2-2-2-2-3-4-GAC-TAC-TGGGGCCAAGGGACCACCGTGACCGTCTCC 1: G/D = 20%, E/V/S = 10%, A/P/R/L/T/Y=5%; 2: G/Y/S=15%, A/D/T/R/P/L/V/N/W/F/I/E = 4,6%; 3: G/A/Y = 20%, P/W/S/D/T = 8%; 4: F = 46%, L/M = 15%, G/I/Y = 8%. |
146 | fdseqlong | GACGTTAGTAAATGAATTTTCTGTATGAGG |
SEQ ID NO: | 描述 | 序列 |
147 | 共用輕鏈文庫 Vk3 _20 /VH3 _23之pRJH110文庫模板;完全Fab編碼區,其輕鏈包含PelB前導序列+Vk3_20 κ V結構域+CL恆定域且包括標籤之重鏈包含PelB+VH3_23V結構域+CH1恆定域 | ATGAAATACCTATTGCCTACGGCAGCCGCTGGATTGTTATTACTCGCGGCCCAGCCGGCCATGGCCGAAATCGTGTTAACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCTTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGAGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGATCCGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCAGTATGGTAGCTCACCGCTGACGTTCGGCCAGGGGACCAAAGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTGGAGCCGCACATCACCATCACCATCACGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCATAATAAGGCGCGCCAATTCTATTTCAAGGAGACAGTCATATGAAATACCTGCTGCCGACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCGAGGTGCAATTGCTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAACCGTTTCCGTATTTTGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGCCGCGGCA |
SEQ ID NO: | 描述 | 序列 |
148 | Fab輕鏈Vk3_20之核苷酸序列 | GAAATCGTGTTAACGCAGTCTCCAGGCACCCTGTCTTTGTCTCCAGGGGAAAGAGCCACCCTCTCTTGCAGGGCCAGTCAGAGTGTTAGCAGCAGCTACTTAGCCTGGTACCAGCAGAAACCTGGCCAGGCTCCCAGGCTCCTCATCTATGGAGCATCCAGCAGGGCCACTGGCATCCCAGACAGGTTCAGTGGCAGTGGATCCGGGACAGACTTCACTCTCACCATCAGCAGACTGGAGCCTGAAGATTTTGCAGTGTATTACTGTCAGCAGTATGGTAGCTCACCGCTGACGTTCGGCCAGGGGACCAAAGTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTGGAGCCGCACATCACCATCACCATCACGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCA |
149 | Fab輕鏈Vk3_20 | EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGAAHHHHHHGAADYKDDDDKGAA |
150 | Fab重鏈VH3_23之核苷酸序列 | GAGGTGCAATTGCTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAACCGTTTCCGTATTTTGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGCCGCGGCA |
151 | Fab重鏈VH3_23 (DP47) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKPFPYFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDAAAEQKLISEEDLNAAA |
SEQ ID NO: | 引子名稱 | 引子序列5 ' - 3 ' |
152 | MS64 | ACGTTCGGCCAGGGGACCAAAGTGG |
153 | DP47CDR3_ba (mod.) | CGCACAGTAATATACGGCCGTGTCC |
154 | DP47-v4-4 | CGAGGACACGGCCGTATATTACTGTGCG-5-1-2-2-3-4-GAC-TAC-TGGGGCCAAGGAACCCTGGTCACCGTCTCG |
155 | DP47-v4-6 | CGAGGACACGGCCGTATATTACTGTGCG-5-1-2-2-2-2-3-4-GAC-TAC- TGGGGCCAAGGAACCCTGGTCACCGTCTCG |
156 | DP47-v4-8 | CGAGGACACGGCCGTATATTACTGTGCG-5-1-2-2-2-2-2-2-3-4-GAC-TAC- TGGGGCCAAGGAACCCTGGTCACCGTCTCG |
157 | fdseqlong | GACGTTAGTAAATGAATTTTCTGTATGAGG |
SEQ ID NO: | 描述 | 序列 |
158 | λ -DP47 文庫Vl3 _19 /VH3 _23之pRJH53文庫模板;完全Fab編碼區,其輕鏈包含PelB前導序列+Vl3_19 λ V結構域+CL恆定域且包括標籤之重鏈包含PelB + VH3_23 V結構域+CH1恆定域 | ATGAAATACCTATTGCCTACGGCAGCCGCTGGATTGTTATTACTCGCGGCCCAGCCGGCCATGGCCTCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCGTGATAGTAGCGGTAATCATGTGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGACAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAATTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGCGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCATAATAAGGCGCGCCAATTCTATTTCAAGGAGACAGTCATATGAAATACCTGCTGCCGACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCGAGGTGCAATTGCTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAACCGTTTCCGTATTTTGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAAGCACTAGTGCCCATCACCATCACCATCACGCCGCGGCA |
SEQ ID NO: | 描述 | 序列 |
159 | Fab輕鏈Vl3_19之核苷酸序列 | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCGTGATAGTAGCGGTAATCATGTGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGACAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAATTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGCGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCA |
160 | Fab輕鏈Vl3_19 | SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSRDSSGNHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSGAAEQKLISEEDLNGAADYKDDDDKGAA |
150 | Fab重鏈VH3_23之核苷酸序列 | 參看表7 |
151 | Fab重鏈VH3_23 (DP47) | 參看表7 |
SEQ ID NO: | 引子名稱 | 引子序列5 ' - 3 ' |
161 | LMB3 | CAGGAAACAGCTATGACCATGATTAC |
162 | Vl_3_19_L3r_V | GGACGGTCAGCTTGGTCCCTCCGCCGAATAC V HV A TT A CC G CT A CT A TC A CG GGAGTTACAGTAATAGTCAGCCTCATCTTCCGC 帶下劃線的:60%初始鹼基及40%隨機分組為M 加粗及斜體:60%初始鹼基及40%隨機分組為N |
163 | Vl_3_19_L3r_HV | GGACGGTCAGCTTGGTCCCTCCGCCGAATAC C MM A TG A TT A CC G CT A CT A TC A CG GGAGTTACAGTAATAGTCAGCCTCATCTTCCGC 帶下劃線的:60%初始鹼基及40%隨機分組為M 加粗及斜體:60%初始鹼基及40%隨機分組為N |
164 | Vl_3_19_L3r_HLV | GGACGGTCAGCTTGGTCCCTCCGCCGAATAC R HM V WG A TG A TT A CC G CT A CT A TC A CG GGAGTTACAGTAATAGTCAGCCTCATCTTC CGC 帶下劃線的:60%初始鹼基及40%隨機分組為M 加粗及斜體:60%初始鹼基及40%隨機分組為N |
165 | RJH80 | TTCGGCGGAGGGACCAAGCTGACCGTCC |
純系 | SEQ ID NO: | 序列 |
8H9 | 166 (VL) | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGC CAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTAT GGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGT AACTCCCGTGTTATGCCTCATAATCGCGTATTCGGCGGAGGGACCAAGCTGACCGTC |
167 (VH) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGC AGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCA GCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGT GTTTTCTACCGTGGTGGTGTTTCTATGGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGT | |
49B4 | 168 (VL) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGTATAGTTCGCAGCCGTATACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
169 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGA GAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA | |
1G4 | 170 (VL) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGTATATTTCGTATTCCATGTTGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
171 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGA GAATACGGTTCTATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA | |
20B7 | 172 (VL) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGTATCAGGCTTTTTCGCTTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
173 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGA GTTAACTACCCGTACTCTTACTGGGGTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA | |
CLC-563 | 174 (VL) | GAGATCGTGCTGACCCAGAGCCCCGGCACACTCTCCCTGTCTCCTGGGGAAAGGGCCACCCTTTCATGC AGAGCCAGCCAGTCCGTCTCTAGTAGCTACCTGGCATGGTATCAGCAGAAGCCAGGACAAGCCCCCCGCCTCCTGATTTAC GGCGCTTCCTCTCGGGCAACTGGTATCCCTGACAGGTTCTCAGGGAGCGGAAGCGGAACAGATTTTACCTTGACTATTTCTAGACTGGAGCCAGAGGACTTCGCCGTGTATTACTGT CAGCAGTACGGTAGTAGCCCCCTCACCTTTGGCCAGGGGACAAAAGTCGAAATCAAG |
175 (VH) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGC AGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCA GCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCTT GACGTTGGTGCTTTCGACTACTGGGGCCAAGGAGCCCTGGTCACCGTCTCGAGT | |
CLC-564 | 176 (VL) | GAGATCGTGCTGACCCAGAGCCCCGGCACACTCTCCCTGTCTCCTGGGGAAAGGGCCACCCTTTCATGC AGAGCCAGCCAGTCCGTCTCTAGTAGCTACCTGGCATGGTATCAGCAGAAGCCAGGACAAGCCCCCCGCCTCCTGATTTAC GGCGCTTCCTCTCGGGCAACTGGTATCCCTGACAGGTTCTCAGGGAGCGGAAGCGGAACAGATTTTACCTTGACTATTTCTAGACTGGAGCCAGAGGACTTCGCCGTGTATTACTGT CAGCAGTACGGTAGTAGCCCCCTCACCTTTGGCCAGGGGACAAAAGTCGAAATCAAG |
177 (VH) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGC AGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCA GCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGTTC GACGTTGGTCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGT | |
17A9 | 178 (VL) | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGC CAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTAT GGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGT AACTCCCGTGTTATGCCTCATAATCGCGTATTCGGCGGAGGGACCAAGCTGACCGTC |
179 (VH) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGC AGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCA GCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGT GTTTTCTACCGTGGTGGTGTTTCTATGGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGT |
純系 | SEQ ID No. | 序列 |
8B9 | 180 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATTTGACGTATTCGCGGTTTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
181 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACGGTTGGATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
182 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYLTYSRFTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
183 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYGWMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
49B4 | 184 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAGTTCGCAGCCGTATACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
185 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
186 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYSSQPYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
187 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
1G4 | 188 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATATTTCGTATTCCATGTTGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
189 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACGGTTCTATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
190 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYISYSMLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
191 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYGSMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
20B7 | 192 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATCAGGCTTTTTCGCTTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
193 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGTTAACTACCCGTACTCTTACTGGGGTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
194 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYQAFSLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
195 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARVNYPYSYWGDFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
CLC-563 | 196 (核苷酸序列輕鏈) | GAGATCGTGCTGACCCAGAGCCCCGGCACACTCTCCCTGTCTCCTGGGGAAAGGGCCACCCTTTCATGCAGAGCCAGCCAGTCCGTCTCTAGTAGCTACCTGGCATGGTATCAGCAGAAGCCAGGACAAGCCCCCCGCCTCCTGATTTACGGCGCTTCCTCTCGGGCAACTGGTATCCCTGACAGGTTCTCAGGGAGCGGAAGCGGAACAGATTTTACCTTGACTATTTCTAGACTGGAGCCAGAGGACTTCGCCGTGTATTACTGTCAGCAGTACGGTAGTAGCCCCCTCACCTTTGGCCAGGGGACAAAAGTCGAAATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
197 (核苷酸序列重鏈) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCTTGACGTTGGTGCTTTCGACTACTGGGGCCAAGGAGCCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
198 (輕鏈) | EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
199 (重鏈) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCALDVGAFDYWGQGALVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
CLC-564 | 200 (核苷酸序列輕鏈) | GAGATCGTGCTGACCCAGAGCCCCGGCACACTCTCCCTGTCTCCTGGGGAAAGGGCCACCCTTTCATGCAGAGCCAGCCAGTCCGTCTCTAGTAGCTACCTGGCATGGTATCAGCAGAAGCCAGGACAAGCCCCCCGCCTCCTGATTTACGGCGCTTCCTCTCGGGCAACTGGTATCCCTGACAGGTTCTCAGGGAGCGGAAGCGGAACAGATTTTACCTTGACTATTTCTAGACTGGAGCCAGAGGACTTCGCCGTGTATTACTGTCAGCAGTACGGTAGTAGCCCCCTCACCTTTGGCCAGGGGACAAAAGTCGAAATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
201 (核苷酸序列重鏈) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGTTCGACGTTGGTCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
202 (輕鏈) | EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
203 (重鏈) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAFDVGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
17A9 | 204 (核苷酸序列輕鏈) | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCGTGTTATGCCTCATAATCGCGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAACTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGC |
205 (核苷酸序列重鏈) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGTGTTTTCTACCGTGGTGGTGTTTCTATGGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
206 (輕鏈) | SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSRVMPHNRVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS | |
207 (重鏈) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARVFYRGGVSMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
純系 | 產量[mg/l] | 單體 [%] | CE-SDS (非紅色) | CE-SDS (紅色) |
8H9 P329GLALA IgG1 | 7 | 100 | 1.2% (176kDa) 96.1% (158kDa) 1.3% (142kDa) | 66.9% (54kDa) 28.9% (25kDa) |
49B4 P329GLALA IgG1 | 7.5 | 100 | 99% (163kDa) 1% (149kDa) | 81% (61.7kDa) 18% (28.9kDa) |
1G4 P329GLALA IgG1 | 1 | 100 | 98.9% (167.4kDa) 1.1% (151kDa) | 80% (63.4kDa) 19% (28.9kDa) |
20B7 P329GLALA IgG1 | 17 | 93 | 97.9% (174kDa) | 79.8% (65.4kDa) 19.9% (29.5kDa) |
CLC-563 P329GLALA IgG1 | 6.2 | 100 | 97.7% (160kDa) | 77.7% (60kDa) 19.8% (26.4kDa) |
CLC-564 P329GLALA IgG1 | 13.5 | 100 | 98.4% (155kDa) | 79.3% (60.1kDa) 19.8% (26.5kDa) |
17A9 P329GLALA IgG1 | 7.5 | 100 | 98.6% (175kDa) 1.4% (153kDa) | 74.1% (61kDa) 25.5% (38kDa) |
SEQ ID NO: | 抗原 | 序列 |
208 | 人類OX40 His | 核苷酸序列 |
209 | 人類OX40 His | LHCVGDTYPSNDRCCHECRPGNGMVSRCSRSQNTVCRPCGPGFYNDVVSSKPCKPCTWCNLRSGSERKQLCTATQDTVCRCRAGTQPLDSYKPGVDCAPCPPGHFSPGDNQACKPWTNCTLAGKHTLQPASNSSDAICEDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVDEQLYFQGGSGLNDIFEAQKIEWHEARAHHHHHH |
210 | 鼠類OX40 His | 核苷酸序列 |
211 | 鼠類OX40 His | TARRLNCVKHTYPSGHKCCRECQPGHGMVSRCDHTRDTLCHPCETGFYNEAVNYDTCKQCTQCNHRSGSELKQNCTPTQDTVCRCRPGTQPRQDSGYKLGVDCVPCPPGHFSPGNNQACKPWTNCTLSGKQTRHPASDSLDAVCEDRSLLATLLWETQRPTFRPTTVQSTTVWPRTSELPSPPTLVTPEGPVDEQLYFQGGSGLNDIFEAQKIEWHEARAHHHHHH |
純系 | 重組人類OX40 (親合力型式) | 重組人類OX40 Fc(kih) (親和力型式) | 重組人類OX40 His (親和力型式) | ||||
ka (1/Ms) | kd (1/s) | KD (M) | ka (1/Ms) | kd (1/s) | KD (M) | ||
8H9 | ++++ | 1.6E+05 | 4.5E-03 | 2.8E-08 | 6.5E+04 | 2.0E-03 | 3.1E-08 |
49B4 | ++ | 2.5E+05 | 1.3E-01 | 5.1E-07 | 1.4E+06 | 6.7E-01 | 4.6E-07 |
1G4 | ++ | 3.0E+05 | 8.4E-08 | 2.8E-07 | 2.3E+06 | 5.7E-01 | 2.5E-07 |
20B7 | +++ | 3.2E+04 | 1.3E-03 | 4.2E-08 | 1.2E+05 | 6.6E-04 | 5.6E-09 |
CLC-563 | ++ | 3.6E+04 | 3.2E-03 | 8.9E-08 | 4.0E+04 | 3.6E-03 | 8.9E-08 |
CLC-564 | ++++ | 3.2E+04 | 4.2E-03 | 1.3E-07 | 3.8E+05 | 5.3E-03 | 1.4E-08 |
純系 | EC 50[nM] |
8H9 | 0.59 |
CLC563 | 1.59 |
20B7 | 1.64 |
49B4 | 4.19 |
CLC-564 | 4.63 |
1G4 | n.a. |
純系 | 來源 | 重組鼠類OX40 (親合力型式) | 重組鼠類OX40 (親和力型式) | ||
ka (1/Ms) | kd (1/s) | KD (M) | |||
20B7 | 噬菌體呈現 | ++ | 4.9E+04 | 1.8E-02 | 3.6E-07 |
純系 | EC 50[nM] |
8H9 | n.d. |
CLC563 | 1.41 |
20B7 | 1.52 |
49B4 | n.d. |
CLC-564 | 3.50 |
1G4 | 48.20 |
純系 | 來源 | 重組獼猴OX40 (親合力型式) | 重組獼猴OX40 (親和力型式) | ||
ka (1/Ms) | kd (1/s) | KD (M) | |||
8H9 | 噬菌體呈現 | ++++ | 1.4E+05 | 9.6E-02 | 6.7E-07 |
20B7 | 噬菌體呈現 | +++ | 1.57E+04 | 1.66E-02 | 1.1E-06 |
49B4 | 噬菌體呈現 | ++ | 1.1E+05 | 3.8E-02 | 3.5E-07 |
1G4 | 噬菌體呈現 | + | 太低而無法偵測 | ||
CLC-563 | 噬菌體呈現 | +++ | 2.8E+04 | 6.9E-04 | 2.5E-08 |
CLC-564 | 噬菌體呈現 | +++ | 2.1E+04 | 7.2E-04 | 3.4E-08 |
SEQ ID NO: | 抗原 | 序列 |
212 | 核苷酸序列二聚人類OX40抗原Fc | CTGCACTGCGTGGGCGACACCTACCCCAGCAACGACCGGTGCTGCCACGAGTGCAGACCCGGCAACGGCATGGTGTCCCGGTGCAGCCGGTCCCAGAACACCGTGTGCAGACCTTGCGGCCCTGGCTTCTACAACGACGTGGTGTCCAGCAAGCCCTGCAAGCCTTGTACCTGGTGCAACCTGCGGAGCGGCAGCGAGCGGAAGCAGCTGTGTACCGCCACCCAGGATACCGTGTGCCGGTGTAGAGCCGGCACCCAGCCCCTGGACAGCTACAAACCCGGCGTGGACTGCGCCCCTTGCCCTCCTGGCCACTTCAGCCCTGGCGACAACCAGGCCTGCAAGCCTTGGACCAACTGCACCCTGGCCGGCAAGCACACCCTGCAGCCCGCCAGCAATAGCAGCGACGCCATCTGCGAGGACCGGGATCCTCCTGCCACCCAGCCTCAGGAAACCCAGGGCCCTCCCGCCAGACCCATCACCGTGCAGCCTACAGAGGCCTGGCCCAGAACCAGCCAGGGGCCTAGCACCAGACCCGTGGAAGTGCCTGGCGGCAGAGCCGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGCTACCCATACGATGTTCCAGATTACGCT |
213 | 二聚人類OX40抗原Fc | LHCVGDTYPSNDRCCHECRPGNGMVSRCSRSQNTVCRPCGPGFYNDVVSSKPCKPCTWCNLRSGSERKQLCTATQDTVCRCRAGTQPLDSYKPGVDCAPCPPGHFSPGDNQACKPWTNCTLAGKHTLQPASNSSDAICEDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGYPYDVPDYA |
純系 | 來源 | 第一次注射 | 第二次注射(抗Ox40純系) | 配位體阻斷 |
8H9 | 噬菌體呈現 | 人類OX40 Fc | 未結合 | 是 |
20B7 | 噬菌體呈現 | 人類OX40 Fc | 結合 | 否 |
1G4 | 噬菌體呈現 | 人類OX40 Fc | 未結合 | 是 |
49B4 | 噬菌體呈現 | 人類OX40 Fc | 未結合 | 是 |
CLC-564 | 噬菌體呈現 | 人類OX40 Fc | 未結合 | 是 |
CLC-564 | 噬菌體呈現 | 人類OX40 Fc | 未結合 | 是 |
純系 | EC 50[nM] |
8H9 | 0.66 |
CLC563 | 1.69 |
20B7 | 2.27 |
49B4 | 2.42 |
CLC-564 | 3.23 |
1G4 | 3.59 |
純系 | EC 50[nM] | SEM (+/-) |
8H9 | 0.003 | 0.001 |
20B7 | 0.090 | 0.015 |
CLC-563 | 0.114 | 0.018 |
CLC-564 | 0.202 | 0.053 |
49B4 | 0.591 | 0.237 |
1G4 | 0.697 | 0.278 |
SEQ ID NO: | 描述 | 序列 |
214 | (8B9) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACGGTTGGATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
180 | VLCL-輕鏈1 (8B9) (核苷酸序列) | 參看表13 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | GAGATCGTGCTGACCCAGTCTCCCGGCACCCTGAGCCTGAGCCCTGGCGAGAGAGCCACCCTGAGCTGCAGAGCCAGCCAGAGCGTGAGCCGGAGCTACCTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCATCGGCGCCAGCACCCGGGCCACCGGCATCCCCGATAGATTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACCATCAGCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCCAGGTGATCCCCCCCACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGAGCTCCGCTAGCACCAAGGGCCCCTCCGTGTTTCCTCTGGCCCCCAGCAGCAAGAGCACCTCTGGCGGAACAGCCGCCCTGGGCTGCCTGGTGAAAGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACTCTGGCGCCCTGACCAGCGGCGTGCACACCTTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGAC |
216 | (8B9) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYGWMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
182 | VLCL -輕鏈1 (8B9) | 參看表13 |
217 | VLCH1-輕鏈2 (28H1) | EIVLTQSPGTLSLSPGERATLSCRASQSVSRSYLAWYQQKPGQAPRLLIIGASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGQVIPPTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD |
218 | (49B4) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
184 | VLCL -輕鏈1 (49B4) (核苷酸序列) | 參看表13 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
219 | (49B4) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
186 | VLCL -輕鏈1 (49B4) | 參看表13 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
220 | (1G4) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACGGTTCTATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
188 | VLCL -輕鏈1 (1G4) (核苷酸序列) | 參看表13 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
221 | (1G4) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYGSMDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
190 | VLCL -輕鏈1 (1G4) | 參看表13 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
222 | (20B7) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGTTAACTACCCGTACTCTTACTGGGGTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
192 | VLCL -輕鏈1 (20B7) (核苷酸序列) | 參看表13 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
223 | (20B7) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARVNYPYSYWGDFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
194 | VLCL -輕鏈1 (20B7) | 參看表13 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
224 | (CLC-563) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCTTGACGTTGGTGCTTTCGACTACTGGGGCCAAGGAGCCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
196 | VLCL -輕鏈1 (CLC-563) (核苷酸序列) | 參看表13 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
225 | (CLC-563) VHCH1-重鏈-(28H1) VHCL | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCALDVGAFDYWGQGALVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
198 | VLCL -輕鏈1 (CLC-563) | 參看表13 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
226 | (CLC-564) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGTTCGACGTTGGTCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
200 | VLCL -輕鏈1 (CLC-564) (核苷酸序列) | 參看表13 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
227 | (CLC-564) VHCH1-重鏈-(28H1) VHCL | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAFDVGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
202 | VLCL -輕鏈1 (CLC-564) | 參看表13 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
純系 | 產量[mg/l] | 單體[%] | CE-SDS (非紅色) | CE-SDS (紅色) |
8H9/FAP P329GLALA IgG1 2+2 | 58 | 100 | 95.3% (254kDa) 3% (237kDa) | 3.2% (114kDa) 71.3% (90.7kDa) 13.3% (28.9kDa) 11.9% (26.2kDa) |
49B4/FAP P329GLALA IgG1 2+2 | 17 | 99 | 98.9% (253kDa) | 3.% (116kDa) 71.4% (92kDa) 12.9% (28.9kDa) 12.1% (25.7kDa) |
1G4/FAP P329GLALA IgG1 2+2 | 0.5 | 99.1 | 93.9% (234kDa) 3.2% (242kDa) 1.2% (244kDa) | 55.5% (90.6kDa) 20.7% (27kDa) 21.6% (25kDa) |
20B7/FAP P329GLALA IgG1 2+2 | 14 | 97.2 | 91.5% (244kDa) 2.3% (227kDa) 1.4% (218kDa) 1.5% (202kDa) | 54.1% (89kDa) 19% (27kDa) 25% (24kDa) |
SEQ ID NO: | 描述 | 序列 |
228 | (28H1) VHCL-重鏈臼 (核苷酸序列) | GAAGTGCAGCTGCTGGAATCCGGCGGAGGCCTGGTGCAGCCTGGCGGATCTCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCTCCCACGCCATGTCCTGGGTCCGACAGGCTCCTGGCAAAGGCCTGGAATGGGTGTCCGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGACTCTGTGAAGGGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAAGGGCTGGCTGGGCAACTTCGACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGCGCTAGCGTGGCCGCTCCCAGCGTGTTCATCTTCCCACCCAGCGACGAGCAGCTGAAGTCCGGCACAGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAATCCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
215 | (28H1) VLCH1-輕鏈2 (核苷酸序列) | 參看表25 |
229 | (28H1) VHCL-重鏈臼 | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
217 | (28H1) VLCH1-輕鏈2 | 參看表25 |
302 | (8H9) VHCH1-重鏈杵 (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACGGTTGGATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
180 | (8H9) VLCL-輕鏈1 (核苷酸序列) | 參看表13 |
303 | (8H9) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYGWMDYWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
182 | (8H9) VLCL-輕鏈1 | 參看表13 |
230 | (49B4) VHCH1-重鏈杵 (核苷酸序列) | AGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
184 | (49B4) VLCL-輕鏈1 (核苷酸序列) | 參看表13 |
231 | (49B4) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
186 | (49B4) VLCL-輕鏈1 | 參看表13 |
232 | (1G4) VHCH1-重鏈杵 (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACGGTTCTATGGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
188 | (1G4) VLCL-輕鏈1 (核苷酸序列) | 參看表13 |
233 | (1G4) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYGSMDYWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
190 | (1G4) VLCL-輕鏈1 | 參看表13 |
234 | (20B7) VHCH1-重鏈杵 (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGTTAACTACCCGTACTCTTACTGGGGTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
192 | (20B7) VLCL-輕鏈1 (核苷酸序列) | 參看表13 |
235 | (20B7) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARVNYPYSYWGDFDYWGQGTTVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
194 | (20B7) VLCL-輕鏈1 | 參看表13 |
236 | (CLC-563) VHCH1-重鏈杵 (核苷酸序列) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCTTGACGTTGGTGCTTTCGACTACTGGGGCCAAGGAGCCCTGGTCACCGTCTCGAGT GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
196 | (CLC-563) VLCL-輕鏈1 (核苷酸序列) | 參看表13 |
237 | (CLC-563) VHCH1-重鏈杵 | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCALDVGAFDYWGQGALVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
198 | (CLC-563) VLCL-輕鏈1 | 參看表13 |
238 | (CLC-564) VHCH1-重鏈杵 (核苷酸序列) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGTTCGACGTTGGTCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGT GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
200 | (CLC-564) VLCL-輕鏈1 (核苷酸序列) | 參看表13 |
239 | (CLC-564) VHCH1-重鏈杵 | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAFDVGPFDYWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
202 | (CLC-564) VLCL-輕鏈1 | 參看表13 |
純系 | 產量[mg/l] | 單體[%] | CE-SDS (非紅色) | CE-SDS (紅色) |
8H9/FAP P329GLALA IgG1 1+1 | 16.5 | 100 | 92.1% (164kDa) 1.9% (145kDa) 3.6% (120.1kDa) | 67.7% (63.6kDa) 13.3% (28.5kDa) 16.5% (25.7kDa) |
1G4/FAP P329GLALA IgG1 1+1 | 12.5 | 98.5 | 85.2% (157kDa) 7.4% (151kDa) 2.8% (139.5kDa) | 69.5% (64.2kDa) 13.1% (28.8kDa) 16.7% (26.2kDa) |
49B4/FAP P329GLALA IgG1 1+1 | 2.3 | 97.9 | 80% (153kDa) 11.9% (141kDa) 4.3% (120kDa) | 70.4% (63.5kDa) 14.7% (28kDa) 13.7% (25kDa) |
20B7/FAP P329GLALA IgG1 1+1 | 22 | 100 | 97.5% (166kDa) 1.3% (149kDa) | 82.7% (56.2kDa) 8.2% (27.2kDa) 8.1% (24.3kDa) |
純系 | 型式 | FAP +細胞 EC 50[nM] | OX40 +細胞 EC 50[nM] |
8H9 (WM266-4) | hu IgG1 | n.a. | 0.59 |
FAP 1+1 | 5.99 | 8.20 | |
FAP 2+2 | 2.88 | 0.93 | |
1G4 (NIH/3T3 huFAP純系39) | hu IgG1 | n.a. | n.a. |
FAP 1+1 | 3.55 | 49.07 | |
FAP 2+2 | 0.77 | 9.37 |
SEQ ID NO: | 描述 | 序列 |
184 | (49B4) VLCL-輕鏈 (核苷酸序列) | 參看表13 |
304 | (49B4) VHCH1 Fc杵VH (4B9) (重鏈1之核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGAGGATCCGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
305 | (49B4) VHCH1 Fc臼VL (4B9) (重鏈2之核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATAC TACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACG TACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGTGGATCTGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
186 | (49B4) VLCL-輕鏈 | 參看表13 |
306 | (49B4) VHCH1 Fc杵VH (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
307 | (49B4) VHCH1 Fc臼VL (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
184 | (49B4) VLCL-輕鏈 (核苷酸序列) | 參看表13 |
308 | (49B4) VHCH1 Fc杵VH (28H1) (核苷酸序列,重鏈1) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGAGGAGGGGGAAGTGGCGGCGGAGGATCTGAGGTGCAGCTGCTGGAATCCGGCGGAGGCCTGGTGCAGCCTGGCGGATCTCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCTCCCACGCCATGTCCTGGGTCCGACAGGCTCCTGGCAAAGGCCTGGAATGGGTGTCCGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGACTCTGTGAAGGGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAAGGGCTGGCTGGGCAACTTCGACTACTGGGGCCAGGGCACCCTGGTCACCGTGTCCAGC |
309 | (49B4) VHCH1 Fc杵VL (28H1) (核苷酸序列,重鏈2) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGAGCCTGAGCCCTGGCGAGAGAGCCACCCTGAGCTGCAGAGCCAGCCAGAGCGTGAGCCGGAGCTACCTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCATCGGCGCCAGCACCCGGGCCACCGGCATCCCCGATAGATTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACCATCAGCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCCAGGTGATCCCCCCCACCTTCGGCCAGGGCACCAAGGTGGAAATCAAG |
186 | (49B4) VLCL-輕鏈 | 參看表13 |
310 | (49B4) VHCH1 Fc杵VH (28H1) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSS |
311 | (49B4) VHCH1 Fc臼VL (28H1) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVSRSYLAWYQQKPGQAPRLLIIGASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGQVIPPTFGQGTKVEIK |
184 | (49B4) VLCL-輕鏈 (核苷酸序列) | 參看表13 |
312 | (49B4) VHCH1 Fc杵VH (DP47) (核苷酸序列,重鏈1) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGAGGAGGGGGAAGTGGCGGCGGAGGATCTGAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCAGCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAAGGCAGCGGATTTGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGC |
313 | (49B4) VHCH1 Fc臼VL (DP47) (核苷酸序列,重鏈2) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAGAATACTACCGTGGTCCGTACGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGAGGCGGCGGAAGCGGAGGGGGAGGCTCTGAAATTGTGCTGACCCAGAGCCCCGGCACCCTGTCACTGTCTCCAGGCGAAAGAGCCACCCTGAGCTGCAGAGCCAGCCAGAGCGTGTCCAGCTCTTACCTGGCCTGGTATCAGCAGAAGCCCGGACAGGCCCCCAGACTGCTGATCTACGGCGCCTCTTCTAGAGCCACCGGCATCCCCGATAGATTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACAATCAGCAGACTGGAACCCGAGGACTTTGCCGTGTATTACTGCCAGCAGTACGGCAGCAGCCCCCTGACCTTTGGCCAGGGCACCAAGGTGGAAATCAAA |
186 | (49B4) VLCL-輕鏈 | 參看表13 |
314 | (49B4) VHCH1 Fc杵VH (DP47) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGSGFDYWGQGTLVTVSS |
315 | (49B4) VHCH1 Fc臼VL (DP47) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCAREYYRGPYDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPLTFGQGTKVEIK |
純系 | 產量[mg/l] | 單體[%] | CE-SDS (非紅色) | CE-SDS (紅色) |
OX40(49B4) /FAP(28H1) P329GLALA IgG1 2+1 | 8.3 | 97.25 (0.75 HMW) | 100 | 73.7 % (81.6 kDa) 0.13% (30.9 kDa) 26.8% (29.5 kDa) |
OX40(49B4) /FAP(4B9) P329GLALA IgG1 2+1 | 7.85 | 99.25 (0.75 HMW) | 100 | 58.71 % (80.44 kDa) 41.29 % (28.91 kDa) |
OX40(49B4) / DP47 P329GLALA IgG1 2+1 | 5.76 | 97.63 (1.33 HMW) | 100 | 73.47 % (81.39 kDa) 0.11 % (30.67 kDa) 26.42 % (29.42 kDa) |
構築體 | hu FAP K D(M) | mu FAP K D(M) | cyno FAP K D(M) |
OX40(49B4) /FAP(28H1) P329GLALA IgG1 2+1 | 1.9E-08 | 3.3E-10 | 3.1E-08 |
OX40(49B4) /FAP(4B9) P329GLALA IgG1 2+1 | 1.0E-09 | 1.1E-07 | 8.5E-10 |
OX40(49B4) / DP47 P329GLALA IgG1 2+1 | n.d. | n.d. | n.d. |
構築體 | EC 50(nM) |
49B4 IgG1 | 0.87 (0.64-1.2) |
OX40(49B4) /FAP(28H1) P329GLALA IgG1 2+1 | 1.96 (1.22-3.14) |
OX40(49B4) /FAP(4B9) P329GLALA IgG1 2+1 | 0.93 (0.7-1.22) |
OX40(49B4) /FAP(DP47) P329GLALA IgG1 2+1 | 1.28 (0.95-1.72) |
純系 | 型式 | FAP +細胞 EC 50[nM] | OX40 +細胞 EC 50[nM] |
49B4, 28H1 | FAP 1+1 | 1.0 | 123.1 |
FAP 2+1 | 70.8 | 6.5 | |
FAP 2+2 | 1.2 | 6.1 | |
49B4, 4B9 | FAP 2+1 | 4.1 | 9.0 |
49B4, DP47 | FAP 2+1 | 5.99 | 9.0 |
純系 | 型式 | EC 50[nM] | +/- SEM |
1G4 | hu IgG1 | 0.37 | 0.03 |
FAP 1+1 | 2.23 | 0.05 | |
FAP 2+2 | 0.75 | 0.16 |
SEQ ID NO: | 構築體 | 來源 | ECD |
39 | 人類4-1BB ECD | 根據Q07011合成 | aa 24-186 |
240 | 獼猴4-1BB ECD | 自獼猴血液分離 | aa 24-186 |
241 | 鼠類4-1BB ECD | 根據P20334合成 | aa 24-187 |
SEQ ID NO: | 抗原 | 序列 |
124 | 核苷酸序列Fc臼鏈 | 參看表2 |
242 | 核苷酸序列人類4-1BB抗原Fc杵鏈 | CTGCAGGACCCCTGCAGCAACTGCCCTGCCGGCACCTTCTGCGACAACAACCGGAACCAGATCTGCAGCCCCTGCCCCCCCAACAGCTTCAGCTCTGCCGGCGGACAGCGGACCTGCGACATCTGCAGACAGTGCAAGGGCGTGTTCAGAACCCGGAAAGAGTGCAGCAGCACCAGCAACGCCGAGTGCGACTGCACCCCCGGCTTCCATTGTCTGGGAGCCGGCTGCAGCATGTGCGAGCAGGACTGCAAGCAGGGCCAGGAACTGACCAAGAAGGGCTGCAAGGACTGCTGCTTCGGCACCTTCAACGACCAGAAGCGGGGCATCTGCCGGCCCTGGACCAACTGTAGCCTGGACGGCAAGAGCGTGCTGGTCAACGGCACCAAAGAACGGGACGTCGTGTGCGGCCCCAGCCCTGCTGATCTGTCTCCTGGGGCCAGCAGCGTGACCCCTCCTGCCCCTGCCAGAGAGCCTGGCCACTCTCCTCAGGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGAGGCCTGAACGACATCTTCGAGGCCCAGAAGATTGAATGGCACGAG |
243 | 核苷酸序列獼猴4-1BB抗原Fc杵鏈 | TTGCAGGATCTGTGTAGTAACTGCCCAGCTGGTACATTCTGTGATAATAACAGGAGTCAGATTTGCAGTCCCTGTCCTCCAAATAGTTTCTCCAGCGCAGGTGGACAAAGGACCTGTGACATATGCAGGCAGTGTAAAGGTGTTTTCAAGACCAGGAAGGAGTGTTCCTCCACCAGCAATGCAGAGTGTGACTGCATTTCAGGGTATCACTGCCTGGGGGCAGAGTGCAGCATGTGTGAACAGGATTGTAAACAAGGTCAAGAATTGACAAAAAAAGGTTGTAAAGACTGTTGCTTTGGGACATTTAATGACCAGAAACGTGGCATCTGTCGCCCCTGGACAAACTGTTCTTTGGATGGAAAGTCTGTGCTTGTGAATGGGACGAAGGAGAGGGACGTGGTCTGCGGACCATCTCCAGCCGACCTCTCTCCAGGAGCATCCTCTGCGACCCCGCCTGCCCCTGCGAGAGAGCCAGGACACTCTCCGCAGGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGAGGCCTGAACGACATCTTCGAGGCCCAGAAGATTGAATGGCACGAG |
244 | 核苷酸序列鼠類4-1BB抗原Fc杵鏈 | GTGCAGAACAGCTGCGACAACTGCCAGCCCGGCACCTTCTGCCGGAAGTACAACCCCGTGTGCAAGAGCTGCCCCCCCAGCACCTTCAGCAGCATCGGCGGCCAGCCCAACTGCAACATCTGCAGAGTGTGCGCCGGCTACTTCCGGTTCAAGAAGTTCTGCAGCAGCACCCACAACGCCGAGTGCGAGTGCATCGAGGGCTTCCACTGCCTGGGCCCCCAGTGCACCAGATGCGAGAAGGACTGCAGACCCGGCCAGGAACTGACCAAGCAGGGCTGTAAGACCTGCAGCCTGGGCACCTTCAACGACCAGAACGGGACCGGCGTGTGCCGGCCTTGGACCAATTGCAGCCTGGACGGGAGAAGCGTGCTGAAAACCGGCACCACCGAGAAGGACGTCGTGTGCGGCCCTCCCGTGGTGTCCTTCAGCCCTAGCACCACCATCAGCGTGACCCCTGAAGGCGGCCCTGGCGGACACTCTCTGCAGGTCCTGGTCGACGAACAGTTATATTTTCAGGGCGGCTCACCCAAATCTGCAGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAATCCGGAGGCCTGAACGACATCTTCGAGGCCCAGAAGATTGAATGGCACGAG |
128 | Fc臼鏈 | 參看表2 |
245 | 人類4-1BB抗原Fc杵鏈 | LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
246 | 獼猴4-1BB抗原Fc杵鏈 | LQDLCSNCPAGTFCDNNRSQICSPCPPNSFSSAGGQRTCDICRQCKGVFKTRKECSSTSNAECDCISGYHCLGAECSMCEQDCKQGQELTKKGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSATPPAPAREPGHSPQVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
247 | 鼠類4-1BB抗原Fc杵鏈 | VQNSCDNCQPGTFCRKYNPVCKSCPPSTFSSIGGQPNCNICRVCAGYFRFKKFCSSTHNAECECIEGFHCLGPQCTRCEKDCRPGQELTKQGCKTCSLGTFNDQNGTGVCRPWTNCSLDGRSVLKTGTTEKDVVCGPPVVSFSPSTTISVTPEGGPGGHSLQVLVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
SEQ ID NO: | 描述 | 序列 |
132 | pRJH33庫模板DP88-4庫的核苷酸序列;完全Fab編碼區,其輕鏈包含PelB前導序列 + Vk1_5 κ V結構域 + CL恆定域且包括標籤之重鏈包含PelB + VH1_69 V-結構域 + CH1恆定域 | TGAAATACCTATTGCCTACGGCAGCCGCTGGATTGTTATTACTCGCGGCCCAGCCGGCCATGGCCGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTCTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCATAATAAGGCGCGCCAATTCTATTTCAAGGAGACAGTCATATGAAATACCTGCTGCCGACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCCAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGACTATCCCCAGGCGGTTACTATGTTATGGATGCCTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAAGCACTAGTGCCCATCACCATCACCATCACGCCGCGGCA |
SEQ ID NO: | 描述 | 序列 |
133 | Fab輕鏈Vk1_5之核苷酸序列 | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTCTACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGTGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCA |
134 | Fab輕鏈Vk1_5 | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGAAEQKLISEEDLNGAADYKDDDDKGAA |
135 | Fab重鏈VH1_69之核苷酸序列 | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGACTATCCCCAGGCGGTTACTATGTTATGGATGCCTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAAGCACTAGTGCCCATCACCATCACCATCACGCCGCGGCA |
136 | Fab重鏈VH1_69 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARLSPGGYYVMDAWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDAAASTSAHHHHHHAAA |
SEQ ID NO: | 引子名稱 | 引子序列5 ' - 3 ' |
137 | LMB3 | CAGGAAACAGCTATGACCATGATTAC |
138 | Vk1_5_L3r_S | CTCGACTTTGGTGCCCTGGCCAAACGT S BA A TA C GA A TT A TA CTGTTGGCAGTAATAAGTTGCAAAATCAT 帶下劃線的:60%初始鹼基及40%隨機分組為M。 加粗及斜體:60%初始鹼基及40%隨機分組為N |
139 | Vk1_5_L3r_SY | CTCGACTTTGGTGCCCTGGCCAAACGT M HR S GR A TA C GA A TT A TA CTGTTGGCAGTAATAAGTTGCAAAATCAT 帶下劃線的:60%初始鹼基及40%隨機分組為M。 加粗及斜體:60%初始鹼基及40%隨機分組為N |
140 | Vk1_5_L3r_SPY | CTCGACTTTGGTGCCCTGGCCAAACGT M HH M SS S GR A TA C GA A TT A TA CTGTTGGCAGTAATAAGTTGCAAAATCAT 帶下劃線的:60%初始鹼基及40%隨機分組為M。 加粗及斜體:60%初始鹼基及40%隨機分組為N |
141 | RJH31 | ACGTTTGGCCAGGGCACCAAAGTCGAG |
142 | RJH32 | TCTCGCACAGTAATACACGGCGGTGTCC |
143 | DP88-v4-4 | GGACACCGCCGTGTATTACTGTGCGAGA-1-2-2-3-4-GAC-TAC-TGGGGCCAAGGGACCACCGTGACCGTCTCC 1: G/D = 20%, E/V/S = 10%, A/P/R/L/T/Y=5%; 2: G/Y/S=15%, A/D/T/R/P/L/V/N/W/F/I/E = 4,6%; 3: G/A/Y = 20%, P/W/S/D/T = 8%; 4: F = 46%, L/M = 15%, G/I/Y = 8%. |
144 | DP88-v4-6 | GGACACCGCCGTGTATTACTGTGCGAGA-1-2-2-2-2-3-4-GAC-TAC-TGGGGCCAAGGGACCACCGTGACCGTCTCC 1: G/D = 20%, E/V/S = 10%, A/P/R/L/T/Y=5%; 2: G/Y/S=15%, A/D/T/R/P/L/V/N/W/F/I/E = 4,6%; 3: G/A/Y = 20%, P/W/S/D/T = 8%; 4: F = 46%, L/M = 15%, G/I/Y = 8%. |
145 | DP88-v4-8 | GGACACCGCCGTGTATTACTGTGCGAGA-1-2-2-2-2-2-2-3-4-GAC-TAC-TGGGGCCAAGGGACCACCGTGACCGTCTCC 1: G/D = 20%, E/V/S = 10%, A/P/R/L/T/Y=5%; 2: G/Y/S=15%, A/D/T/R/P/L/V/N/W/F/I/E = 4,6%; 3: G/A/Y = 20%, P/W/S/D/T = 8%; 4: F = 46%, L/M = 15%, G/I/Y = 8%. |
146 | fdseqlong | GACGTTAGTAAATGAATTTTCTGTATGAGG |
SEQ ID NO: | 描述 | 序列 |
158 | λ-DP47 文庫 Vl3_19/VH3_23 之pRJH53庫模板 ;完全Fab編碼區,其輕鏈包含PelB前導序列 + Vl3_19 λ V結構域 + CL恆定域且包括標籤之重鏈包含PelB + VH3_23 V結構域 + CH1恆定域 | ATGAAATACCTATTGCCTACGGCAGCCGCTGGATTGTTATTACTCGCGGCCCAGCCGGCCATGGCCTCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCGTGATAGTAGCGGTAATCATGTGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGACAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAATTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGCGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCATAATAAGGCGCGCCAATTCTATTTCAAGGAGACAGTCATATGAAATACCTGCTGCCGACCGCTGCTGCTGGTCTGCTGCTCCTCGCTGCCCAGCCGGCGATGGCCGAGGTGCAATTGCTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAAACCGTTTCCGTATTTTGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAAGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAAGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACGCGGCCGCAAGCACTAGTGCCCATCACCATCACCATCACGCCGCGGCA |
SEQ ID NO: | 描述 | 序列 |
159 | Fab輕鏈Vl3_19之核苷酸序列 | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCGTGATAGTAGCGGTAATCATGTGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGACAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAATTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGCGGAGCCGCAGAACAAAAACTCATCTCAGAAGAGGATCTGAATGGAGCCGCAGACTACAAGGACGACGACGACAAGGGTGCCGCA |
160 | Fab輕鏈Vl3_19 | SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSRDSSGNHVVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSGAAEQKLISEEDLNGAADYKDDDDKGAA |
150 | Fab重鏈VH3_23之核苷酸序列 | 參看表7 |
151 | Fab重鏈VH3_23 (DP47) | 參看表7 |
SEQ ID NO: | 引子名稱 | 引子序列5 ' - 3 ' |
161 | LMB3 | CAGGAAACAGCTATGACCATGATTAC |
162 | Vl_3_19_L3r_V | GGACGGTCAGCTTGGTCCCTCCGCCGAATAC V HV A TT A CC G CT A CT A TC A CG GGAGTTACAGTAATAGTCAGCCTCATCTTCCGC 帶下劃線的:60%初始鹼基及40%隨機分組為M 加粗及斜體:60%初始鹼基及40%隨機分組為N |
163 | Vl_3_19_L3r_HV | GGACGGTCAGCTTGGTCCCTCCGCCGAATAC C MM A TG A TT A CC G CT A CT A TC A CG GGAGTTACAGTAATAGTCAGCCTCATCTTCCGC 帶下劃線的:60%初始鹼基及40%隨機分組為M 加粗及斜體:60%初始鹼基及40%隨機分組為N |
164 | Vl_3_19_L3r_HLV | GGACGGTCAGCTTGGTCCCTCCGCCGAATAC R HM V WG A TG A TT A CC G CT A CT A TC A CG GGAGTTACAGTAATAGTCAGCCTCATCTTC CGC 帶下劃線的:60%初始鹼基及40%隨機分組為M 加粗及斜體:60%初始鹼基及40%隨機分組為N |
165 | RJH80 | TTCGGCGGAGGGACCAAGCTGACCGTCC |
248 | MS63 | TTTCGCACAGTAATATACGGCCGTGTCC |
154 | DP47-v4-4 | CGAGGACACGGCCGTATATTACTGTGCG-5-1-2-2-3-4-GAC-TAC-TGGGGCCAAGGAACCCTGGTCACCGTCTCG |
155 | DP47-v4-6 | CGAGGACACGGCCGTATATTACTGTGCG-5-1-2-2-2-2-3-4-GAC-TAC- TGGGGCCAAGGAACCCTGGTCACCGTCTCG |
156 | DP47-v4-8 | CGAGGACACGGCCGTATATTACTGTGCG-5-1-2-2-2-2-2-2-3-4-GAC-TAC- TGGGGCCAAGGAACCCTGGTCACCGTCTCG |
157 | fdseqlong | GACGTTAGTAAATGAATTTTCTGTATGAGG |
純系 | SEQ ID NO: | 序列 |
12B3 | 249 (VL) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGTATCATTCGTATCCGCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
250 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGA TCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA | |
25G7 | 251(VL) | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGC CAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTAT GGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGT AACTCCCTTGATAGGCGCGGTATGTGGGTATTCGGCGGAGGGACCAAGCTGACCGTC |
252 (VH) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGC AGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCA GCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGT GACGACCCGTGGCCGCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGT | |
11D5 | 253 (VL) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGCTTAATTCGTATCCTCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
254 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGAT CTACTCTGATCTACGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA | |
9B11 | 255 (VL) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGGTTAATTCTTATCCGCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
256 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGA TCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA | |
20G2 | 257 (VL) | GACATCCAGATGACCCAGTCTCCATCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGC CGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTAT GATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGC CAACAGCAGCACTCGTATTATACGTTTGGCCAGGGCACCAAAGTCGAGATCAAG |
258 (VH) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGC AGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGA GGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGA TCTTACTACTGGGAATCTTACCCGTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCAGC |
純系 | SEQ ID No. | 序列 |
12B3 | 259 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATCATTCGTATCCGCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
260 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
261 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYHSYPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
262 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
25G7 | 263 (核苷酸序列輕鏈) | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCTTGATAGGCGCGGTATGTGGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAACTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGC |
264 (核苷酸序列重鏈) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGTGACGACCCGTGGCCGCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
265 (輕鏈) | SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSLDRRGMWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS | |
266 (重鏈) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
11D5 | 267 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGCTTAATTCGTATCCTCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
268 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTACTCTGATCTACGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
269 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQLNSYPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
270 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
9B11 | 271 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGTTAATTCTTATCCGCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
272 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
273 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQVNSYPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
274 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK | |
20G2 | 275 (核苷酸序列輕鏈) | GACATCCAGATGACCCAGTCTCCATCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGCAGCACTCGTATTATACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
276 (核苷酸序列重鏈) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTACTACTGGGAATCTTACCCGTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA | |
277 (輕鏈) | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQQHSYYTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC | |
278 (重鏈) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSYYWESYPFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
純系 | 產量[mg/l] | 單體[%] | CE-SDS (非紅色) | CE-SDS (紅色) |
12B3 P329GLALA IgG1 | 4 | 98 | 98.6% (173kDa) | 22.5% (29kDa) 75.5% (64kDa) |
25G7 P329GLALA IgG1 | 25 | 100 | 99.7% (181.6kDa) | 76.8% (65kDa) 23% (42kDa) |
11D5 P329GLALA IgG1 | 9.7 | 98.7 | 99.6% (176kDa) | tbd. |
9B11 P329GLALA IgG1 | 22 | 100 | 100% (153 kDa) | 2% (127 kDa) 72.3 % (114 kDa) 24.6 % (37.1 kDa) |
20G2 P329GLALA IgG1 | 11 | 100 | 98.5% (166kDa) | 80.2% (62.8kDa) 18% (28.4kDa) |
純系 | 來源 | 充足人類4-1BB (親合力型式) | 充足人類4-1BB (親和力型式) | ||
ka (1/Ms) | kd (1/s) | KD (M) | |||
12B3 | 噬菌體呈現 | ++++ | 3.4E+04 | 1.0E-03 | 3.0E-08 |
25G7 | 噬菌體呈現 | + | 2.9E+04 | 9.9E-04 | 3.4E-08 |
11D5 | 噬菌體呈現 | +++ | 3.2E+04 | 1.2E-03 | 3.6E-08 |
9B11 | 噬菌體呈現 | ++ | 2.7E+04 | 3.9E-03 | 1.4E-07 |
純系 | EC 50[nM] |
25G7 | 29 |
12B3 | 0.95 |
11D5 | 1.46 |
9B11 | 4.485 |
20G2 | n.d. |
純系 | 來源 | 重組鼠類4-1BB (親合力型式) | 重組鼠類4-1BB (親和力型式) | ||
ka (1/Ms) | kd (1/s) | KD (M) | |||
20G2 | 噬菌體呈現 | +++++ | 2.4E+04 | 3.4E-04 | 1.4E-08 |
純系 | EC 50CD8 [nM] | EC 50CD4 [nM] |
25G7 | n.d. | n.d. |
12B3 | n.d. | n.d. |
11D5 | n.d | n.d |
9B11 | n.d | n.d |
20G2 | 16.36 | 10.10 |
純系 | EC 50CD8 [nM] | EC 50CD4 [nM] |
20G2 mu IgG1 κ DAPG | 17.91 | 11.22 |
20G2 mu IgG1 κ wt | 18.51 | 9.917 |
純系 | 來源 | 重組獼猴4-1BB (親合力型式) | 重組獼猴4-1BB (親和力型式) | ||
ka (1/Ms) | kd (1/s) | KD (M) | |||
12B3 | 噬菌體呈現 | +++ | 3.8E+04 | 7.8E-04 | 2.0E-08 |
25G7 | 噬菌體呈現 | ++ | 2.7E+04 | 4.6E-04 | 1.7E-08 |
11D5 | 噬菌體呈現 | +++ | 3.1E+04 | 7.7E-04 | 2.4E-08 |
9B11 | 噬菌體呈現 | + | 2.6E+04 | 2.0E-03 | 7.8E-08 |
純系 | EC 50CD8 [nM] | EC 50CD4 [nM] |
25G7 | 4.52 | 6.68 |
12B3 | 0.47 | 0.59 |
11D5 | 1.04 | 0.97 |
9B11 | n.d. | n.d. |
純系 | 來源 | 第一注射 | 第二注射(抗-4-1BB純系) | 配位體阻斷 |
12B3 | 噬菌體呈現 | 人類4-1BB Fc(kih) | 未結合 | 是 |
25G7 | 噬菌體呈現 | 人類4-1BB Fc(kih) | 結合 | 否 |
11D5 | 噬菌體呈現 | 人類4-1BB Fc(kih) | 未結合 | 是 |
9B11 | 噬菌體呈現 | 人類4-1BB Fc(kih) | 未結合 | 是 |
20G2 | 噬菌體呈現 | 鼠類4-1BB Fc(kih) | 未結合 | 是 |
固定於晶片上 | 第一注射 | 第二注射 | |||
25G7 | 12B3 | 9B11 | 11D5 | ||
9B11 | 人類4-1BB Fc(kih) | 1 | 0 | X | 0 |
11D5 | 人類4-1BB Fc(kih) | 1 | 0 | 0 | X |
融合之結構域 (SEQ ID NO:) | 胺基酸序列 | |
構築體1 | mu4-1BB D1 / hu4-1BB D2 Fc杵 (279) | VQNSCDNCQPGTFCRKYNPVCKSCPPSTFSSIGGQPNCNICRVCAGYFRFKKFCSSTHNAECECIEGFHCLGPQCTRCEKDCRPGQELTKQGCKDCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
Fc臼 (128) | DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK | |
構築體2 | hu4-1BB D1 / mu4-1BB D2 Fc杵 (280) | LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKTCSLGTFNDQNGTGVCRPWTNCSLDGRSVLKTGTTEKDVVCGPPVVSFSPSTTISVTPEGGPGGHSLQVLVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
Fc臼 (128) | DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK | |
構築體3 | hu4-1BB D1 Fc杵 (281) | LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCKVDEQLYFQGGSPKSADKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKSGGLNDIFEAQKIEWHE |
Fc臼 (128) | DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNRFTQKSLSLSPGK |
融合之結構域 (SEQ ID NO:) | 胺基酸序列 | |
構築體1 | 鼠類4-1BB D1 (N端) (282) | VQNSCDNCQPGTFCRKYNPVCKSCPPSTFSSIGGQPNCNICRVCAGYFRFKKFCSSTHNAECECIEGFHCLGPQCTRCEKDCRPGQELTKQGCK |
人類4-1BB D2 (C端) (283) | DCCFGTFNDQKRGICRPWTNCSLDGKSVLVNGTKERDVVCGPSPADLSPGASSVTPPAPAREPGHSPQ | |
構築體2 | 人類4-1BB D1 (N端) (284) | LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCK |
鼠類4-1BB D2 (C端) (285) | TCSLGTFNDQNGTGVCRPWTNCSLDGRSVLKTGTTEKDVVCGPPVVSFSPSTTISVTPEGGPGGHSLQVL | |
構築體3 | 人類4-1BB D1 (N端) (284) | LQDPCSNCPAGTFCDNNRNQICSPCPPNSFSSAGGQRTCDICRQCKGVFRTRKECSSTSNAECDCTPGFHCLGAGCSMCEQDCKQGQELTKKGCK |
純系 | 雜交4-1BB Fc(kih) | ||||
構築體1 | 構築體2 | 構築體3 | |||
鼠類4-1BB D1 (N-) | 人類4-1BB D2 (C-) | 人類4-1BB D1 (N-) | 鼠類4-1BB D2 (C-) | 人類4-1BB D1 (N-) | |
12B3 | 0 | 1 | 1 | ||
25G7 | 1 | 0 | 0 | ||
11D5 | 0 | 1 | 1 | ||
9B11 | 0 | 1 | 1 | ||
20G2 | 1 | 0 | 0 | ||
純系 | EC 50CD8 [nM] |
25G7 | 0.12 |
12B3 | 0.07 |
11D5 | 0.06 |
9B11 | 0.12 |
SEQ ID NO: | 描述 | 序列 |
286 | (12B3) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
259 | VLCL-輕鏈1 (12B3) (核苷酸序列) | 參看表40 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | GAGATCGTGCTGACCCAGTCTCCCGGCACCCTGAGCCTGAGCCCTGGCGAGAGAGCCACCCTGAGCTGCAGAGCCAGCCAGAGCGTGAGCCGGAGCTACCTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCATCGGCGCCAGCACCCGGGCCACCGGCATCCCCGATAGATTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACCATCAGCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCCAGGTGATCCCCCCCACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGAGCTCCGCTAGCACCAAGGGCCCCTCCGTGTTTCCTCTGGCCCCCAGCAGCAAGAGCACCTCTGGCGGAACAGCCGCCCTGGGCTGCCTGGTGAAAGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACTCTGGCGCCCTGACCAGCGGCGTGCACACCTTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGAC |
287 | (12B3) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
261 | VLCL-輕鏈1 (12B3) | 參看表40 |
217 | VLCH1-輕鏈2 (28H1) | EIVLTQSPGTLSLSPGERATLSCRASQSVSRSYLAWYQQKPGQAPRLLIIGASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGQVIPPTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD |
288 | (25G7) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGTGACGACCCGTGGCCGCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
263 | VLCL-輕鏈1 (25G7) (核苷酸序列) | 參看表40 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
289 | (25G7) VHCH1-重鏈-(28H1) VHCL | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
265 | VLCL-輕鏈1 (25G7) | 參看表40 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
290 | (11D5) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTACTCTGATCTACGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
267 | VLCL-輕鏈1 (11D5) (核苷酸序列) | 參看表40 |
215 | VLCH1-輕鏈2 (28H1) (核苷酸序列) | 參看上文 |
291 | (11D5) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
269 | VLCL-輕鏈1 (11D5) | 參看表40 |
217 | VLCH1-輕鏈2 (28H1) | 參看上文 |
292 | (9B11) VHCH1-重鏈-(28H1) VHCL (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCCGTGTTCCCTCTGGCCCCTTCCAGCAAGTCTACCTCTGGCGGCACAGCCGCTCTGGGCTGCCTCGTGAAGGACTACTTCCCCGAGCCTGTGACAGTGTCCTGGAACTCTGGCGCCCTGACATCCGGCGTGCACACCTTTCCAGCTGTGCTGCAGTCCTCCGGCCTGTACTCCCTGTCCTCCGTCGTGACAGTGCCCTCCAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCTCCAACACCAAGGTGGACAAGAAGGTGGAACCCAAGTCCTGCGACAAGACCCACACCTGTCCCCCTTGTCCTGCCCCTGAAGCTGCTGGCGGCCCTAGCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCTCCCGGACCCCCGAAGTGACCTGCGTGGTGGTGGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCTAGAGAGGAACAGTACAACTCCACCTACCGGGTGGTGTCCGTGCTGACCGTGCTGCACCAGGATTGGCTGAACGGCAAAGAGTACAAGTGCAAGGTGTCCAACAAGGCCCTGGGAGCCCCCATCGAAAAGACCATCTCCAAGGCCAAGGGCCAGCCTCGCGAGCCTCAGGTGTACACCCTGCCCCCTAGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGACCTGTCTCGTGAAAGGCTTCTACCCCTCCGATATCGCCGTGGAATGGGAGAGCAACGGCCAGCCCGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACTCCGACGGCTCATTCTTCCTGTACTCTAAGCTGACAGTGGACAAGTCCCGGTGGCAGCAGGGCAACGTGTTCTCCTGCTCCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGTCCCTGTCTCCCGGGGGAGGCGGAGGATCTGGCGGAGGCGGATCCGGTGGTGGCGGATCTGGGGGCGGTGGATCTGAGGTGCAGCTGCTGGAATCTGGGGGAGGACTGGTGCAGCCAGGCGGATCTCTGAGGCTGTCCTGCGCTGCTTCCGGCTTTACCTTCTCCAGCCACGCCATGAGTTGGGTGCGCCAGGCACCCGGAAAAGGACTGGAATGGGTGTCAGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGATAGCGTGAAGGGCCGGTTCACCATCTCTCGGGATAACAGCAAGAATACTCTGTACCTGCAGATGAACTCCCTGCGCGCTGAAGATACCGCTGTGTATTACTGCGCCAAGGGCTGGCTGGGCAACTTCGATTACTGGGGCCAGGGAACCCTCGTGACTGTCTCGAGCGCTTCTGTGGCCGCTCCCTCCGTGTTCATCTTCCCACCTTCCGACGAGCAGCTGAAGTCCGGCACTGCCTCTGTCGTGTGCCTGCTGAACAACTTCTACCCTCGGGAAGCCAAGGTGCAGTGGAAAGTGGATAACGCCCTGCAGTCCGGCAACTCCCAGGAATCCGTGACCGAGCAGGACTCCAAGGACAGCACCTACTCCCTGAGCAGCACCCTGACCCTGTCCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGTGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGTCCTTCAACCGGGGCGAGTGC |
271 | VLCL-輕鏈1 (9B11) (核苷酸序列) | 參看表40 |
215 | VLCH1-輕鏈2 (9B11) (核苷酸序列) | 參看上文 |
293 | (9B11) VHCH1-重鏈-(28H1) VHCL | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
273 | VLCL-輕鏈1 (9B11) | 參看表40 |
217 | VLCH1-輕鏈2 (9B11) | 參看上文 |
純系 | 單體 [%] | 產量[mg/l] |
12B3/FAP P329GLALA IgG1 2+2 | 97.6 | 6.8 |
25G7/FAP P329GLALA IgG1 2+2 | 98.4 | 13 |
11D5/FAP P329GLALA IgG1 2+2 | 100 | 8.7 |
9B11/FAP P329GLALA IgG1 2+2 | 99 | 0.3 |
純系 | EC 50[nM] |
25G7 | 11.14 |
25G7/ FAP 28H1 2+2 | 13.21 |
25G7/ DP47 2+2 | 50.03 |
12B3 | 0.43 |
12B3/ FAP 28H1 2+2 | 1.99 |
11D5 | 1.35 |
11D5/ FAP 28H1 2+2 | 7.91 |
純系 | NIH/3T3-huFAP純系19細胞之EC 50[nM] | WM-266-4之EC 50[nM] |
12B3/ FAP 28H1 2+2 | 12 | n.d. |
25G7/ FAP 28H1 2+2 | 0.4 | n.d. |
11D5/FAP 28H1 2+2 | 0.9 | 0.7 |
25G7/ DP47 2+2 | 0.4 | n.d. |
純系 | NIH/3T3-huFAP純系19之EC 50[nM] |
12B3/ FAP 28H1 2+2 | 0.2 |
11D5/FAP 28H1 2+2 | 0.2 |
25G7/ FAP 28H1 2+2 | 0.1 |
SEQ ID NO: | 描述 | 序列 |
228 | (28H1) VHCL-重鏈臼 (核苷酸序列) | GAAGTGCAGCTGCTGGAATCCGGCGGAGGCCTGGTGCAGCCTGGCGGATCTCTGAGACTGTCCTGCGCCGCCTCCGGCTTCACCTTCTCCTCCCACGCCATGTCCTGGGTCCGACAGGCTCCTGGCAAAGGCCTGGAATGGGTGTCCGCCATCTGGGCCTCCGGCGAGCAGTACTACGCCGACTCTGTGAAGGGCCGGTTCACCATCTCCCGGGACAACTCCAAGAACACCCTGTACCTGCAGATGAACTCCCTGCGGGCCGAGGACACCGCCGTGTACTACTGTGCCAAGGGCTGGCTGGGCAACTTCGACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGCGCTAGCGTGGCCGCTCCCAGCGTGTTCATCTTCCCACCCAGCGACGAGCAGCTGAAGTCCGGCACAGCCAGCGTGGTGTGCCTGCTGAACAACTTCTACCCCCGCGAGGCCAAGGTGCAGTGGAAGGTGGACAACGCCCTGCAGAGCGGCAACAGCCAGGAATCCGTGACCGAGCAGGACAGCAAGGACTCCACCTACAGCCTGAGCAGCACCCTGACCCTGAGCAAGGCCGACTACGAGAAGCACAAGGTGTACGCCTGCGAAGTGACCCACCAGGGCCTGTCCAGCCCCGTGACCAAGAGCTTCAACCGGGGCGAGTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
215 | (28H1) VLCH1-輕鏈2 (核苷酸序列) | GAGATCGTGCTGACCCAGTCTCCCGGCACCCTGAGCCTGAGCCCTGGCGAGAGAGCCACCCTGAGCTGCAGAGCCAGCCAGAGCGTGAGCCGGAGCTACCTGGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCCAGACTGCTGATCATCGGCGCCAGCACCCGGGCCACCGGCATCCCCGATAGATTCAGCGGCAGCGGCTCCGGCACCGACTTCACCCTGACCATCAGCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCCAGGTGATCCCCCCCACCTTCGGCCAGGGCACCAAGGTGGAAATCAAGAGCTCCGCTAGCACCAAGGGCCCCTCCGTGTTTCCTCTGGCCCCCAGCAGCAAGAGCACCTCTGGCGGAACAGCCGCCCTGGGCTGCCTGGTGAAAGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAACTCTGGCGCCCTGACCAGCGGCGTGCACACCTTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCCCTGAGCAGCGTGGTGACAGTGCCCTCCAGCAGCCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGAC |
229 | (28H1) VHCL-重鏈臼 | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSHAMSWVRQAPGKGLEWVSAIWASGEQYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWLGNFDYWGQGTLVTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
217 | (28H1) VLCH1-輕鏈2 | EIVLTQSPGTLSLSPGERATLSCRASQSVSRSYLAWYQQKPGQAPRLLIIGASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGQVIPPTFGQGTKVEIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD |
294 | (12B3) VHCH1-重鏈杵 (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
259 | (12B3) VLCL-輕鏈1 (核苷酸序列) | 參看表40 |
295 | (12B3) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
261 | (12B3) VLCL-輕鏈1 | 參看表40 |
296 | (25G7) VHCH1-重鏈杵 (核苷酸序列) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGTGACGACCCGTGGCCGCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
263 | (25G7) VLCL-輕鏈1 (核苷酸序列) | 參看表40 |
297 | (25G7) VHCH1-重鏈杵 | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
265 | (25G7) VLCL-輕鏈1 | 參看表40 |
298 | (11D5) VHCH1-重鏈杵 (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTACTCTGATCTACGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCA GCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
267 | (11D5) VLCL-輕鏈1 (核苷酸序列) | 參看表40 |
299 | (11D5) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
269 | (11D5) VLCL-輕鏈1 | 參看表40 |
300 | (9B11) VHCH1-重鏈杵 (核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCTAGCGTGTTCCCTCTGGCCCCTAGCAGCAAGAGCACAAGTGGAGGAACAGCCGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAGCCCGTGACCGTGTCCTGGAATTCTGGCGCCCTGACAAGCGGCGTGCACACATTTCCAGCCGTGCTGCAGAGCAGCGGCCTGTACTCTCTGAGCAGCGTCGTGACCGTGCCCTCTAGCTCTCTGGGCACCCAGACCTACATCTGCAACGTGAACCACAAGCCCAGCAACACCAAAGTGGACAAGAAGGTGGAACCCAAGAGCTGCGACAAGACCCACACCTGTCCCCCTTGCCCTGCCCCTGAAGCTGCTGGTGGCCCTTCCGTGTTCCTGTTCCCCCCAAAGCCCAAGGACACCCTGATGATCAGCCGGACCCCCGAAGTGACCTGCGTGGTGGTCGATGTGTCCCACGAGGACCCTGAAGTGAAGTTCAATTGGTACGTGGACGGCGTGGAAGTGCACAATGCCAAGACCAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATGCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGTGGTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTACAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTAAA |
271 | (9B11) VLCL-輕鏈1 (核苷酸序列) | 參看表40 |
301 | (9B11) VHCH1-重鏈杵 | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK |
273 | (9B11) VLCL-輕鏈1 | 參看表40 |
純系 | 產量[mg/l] | 單體[%] |
12B3/FAP P329GLALA IgG1 1+1 | 14 | 94.8 |
25G7/FAP P329GLALA IgG1 1+1 | 33 | 92.2 |
純系 | EC 50[nM] |
25G7 | 11.1 |
25G7/ FAP 28H1 1+1 | > 70 |
25G7/ DP47 1+1 | > 70 |
12B3 | 0.4 |
12B3/ FAP 28H1 1+1 | 8 |
純系 | NIH/3T3-huFAP純系19細胞之EC 50[nM] | WM-266-4細胞之EC 50[nM] |
12B3/ FAP 28H1 1+1 | 2.4 | 1.7 |
25G7/ FAP 28H1 1+1 | 4.4 | 5 |
25G7/ DP47 1+1 | n.d. | n.d. |
純系 | NIH/3T3-huFAP純系19之EC 50[nM] |
12B3/ FAP 28H1 1+1 | 0.9 |
25G7/ FAP 28H1 1+1 | ~2 |
SEQ ID NO: | 描述 | 序列 |
259 | (12B3) VLCL-輕鏈 (核苷酸序列) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATCATTCGTATCCGCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
316 | (12B3) VHCH1 Fc杵VH (4B9) (HC 1之核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTGGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
317 | (12B3) VHCH1 Fc臼VL (4B9) (HC2之核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTGGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
261 | (12B3) VLCL-輕鏈 | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYHSYPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
318 | (12B3) VHCH1 Fc杵VH (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
319 | (12B3) VHCH1 Fc臼VL (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
263 | (25G7) VLCL-輕鏈 (核苷酸序列) | TCGTCTGAGCTGACTCAGGACCCTGCTGTGTCTGTGGCCTTGGGACAGACAGTCAGGATCACATGCCAAGGAGACAGCCTCAGAAGTTATTATGCAAGCTGGTACCAGCAGAAGCCAGGACAGGCCCCTGTACTTGTCATCTATGGTAAAAACAACCGGCCCTCAGGGATCCCAGACCGATTCTCTGGCTCCAGCTCAGGAAACACAGCTTCCTTGACCATCACTGGGGCTCAGGCGGAAGATGAGGCTGACTATTACTGTAACTCCCTTGATAGGCGCGGTATGTGGGTATTCGGCGGAGGGACCAAGCTGACCGTCCTAGGTCAACCCAAGGCTGCCCCCAGCGTGACCCTGTTCCCCCCCAGCAGCGAGGAACTGCAGGCCAACAAGGCCACCCTGGTCTGCCTGATCAGCGACTTCTACCCAGGCGCCGTGACCGTGGCCTGGAAGGCCGACAGCAGCCCCGTGAAGGCCGGCGTGGAGACCACCACCCCCAGCAAGCAGAGCAACAACAAGTACGCCGCCAGCAGCTACCTGAGCCTGACCCCCGAGCAGTGGAAGAGCCACAGGTCCTACAGCTGCCAGGTGACCCACGAGGGCAGCACCGTGGAGAAAACCGTGGCCCCCACCGAGTGCAGC |
320 | (25G7) VHCH1 Fc杵VH (4B9) (核苷酸序列,重鏈1) | GAGGTGCAGCTGCTGGAATCTGGCGGCGGACTGGTGCAGCCTGGCGGATCTCTGAGACTGAGCTGTGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTGCGCCAGGCCCCTGGAAAAGGCCTGGAATGGGTGTCCGCCATCTCTGGCAGCGGCGGCAGCACCTACTACGCCGATTCTGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTATTGCGCCAGGGACGACCCCTGGCCCCCCTTTGATTATTGGGGACAGGGCACCCTCGTGACCGTGTCCAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
321 | (25G7) VHCH1 Fc臼VL (4B9) (核苷酸序列,重鏈2) | GAGGTGCAGCTGCTGGAATCTGGCGGCGGACTGGTGCAGCCTGGCGGATCTCTGAGACTGAGCTGTGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTGCGCCAGGCCCCTGGAAAAGGCCTGGAATGGGTGTCCGCCATCTCTGGCAGCGGCGGCAGCACCTACTACGCCGATTCTGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTATTGCGCCAGGGACGACCCCTGGCCCCCCTTTGATTATTGGGGACAGGGCACCCTCGTGACCGTGTCCAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
265 | (25G7) VLCL-輕鏈 | SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSLDRRGMWVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS |
322 | (25G7) VHCH1 Fc杵VH (4B9) (重鏈1) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
323 | (25G7) VHCH1 Fc臼VL (4B9) (重鏈2) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
267 | (11D5) VLCL-輕鏈 (核苷酸序列) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGCTTAATTCGTATCCTCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
324 | (11D5) VHCH1 Fc杵VH (4B9) (核苷酸序列,重鏈1) | CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACCCGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTTCCGGCGGCACCTTCAGCAGCTACGCCATTTCTTGGGTGCGCCAGGCCCCTGGACAGGGCCTGGAATGGATGGGCGGCATCATCCCCATCTTCGGCACCGCCAACTACGCCCAGAAATTCCAGGGCAGAGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGCGGAGCGAGGACACCGCCGTGTACTACTGTGCCAGAAGCACCCTGATCTACGGCTACTTCGACTACTGGGGCCAGGGCACCACCGTGACCGTGTCTAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
325 | (11D5) VHCH1 Fc臼VL (4B9) (核苷酸序列,重鏈2) | CAGGTGCAGCTGGTGCAGTCTGGCGCCGAAGTGAAGAAACCCGGCAGCAGCGTGAAGGTGTCCTGCAAGGCTTCCGGCGGCACCTTCAGCAGCTACGCCATTTCTTGGGTGCGCCAGGCCCCTGGACAGGGCCTGGAATGGATGGGCGGCATCATCCCCATCTTCGGCACCGCCAACTACGCCCAGAAATTCCAGGGCAGAGTGACCATCACCGCCGACAAGAGCACCAGCACCGCCTACATGGAACTGAGCAGCCTGCGGAGCGAGGACACCGCCGTGTACTACTGTGCCAGAAGCACCCTGATCTACGGCTACTTCGACTACTGGGGCCAGGGCACCACCGTGACCGTGTCTAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
269 | (11D5) VLCL-輕鏈 | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQLNSYPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
326 | (11D5) VHCH1 Fc杵VH (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
327 | (11D5) VHCH1 Fc臼VL (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
271 | (9B11) VLCL-輕鏈 (核苷酸序列) | GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACCGTGTCACCATCACTTGCCGTGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCACGTTTCAGCGGCAGTGGATCCGGGACAGAATTCACTCTCACCATCAGCAGCTTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGTTAATTCTTATCCGCAGACGTTTGGCCAGGGCACCAAAGTCGAGATCAAGCGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT |
328 | (9B11) VHCH1 Fc杵VH (4B9) (核苷酸序列,重鏈1) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
329 | (9B11) VHCH1 Fc臼VL (4B9) (核苷酸序列,重鏈2) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
273 | (9B11) VLCL-輕鏈 | DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYDASSLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQVNSYPQTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC |
330 | (9B11) VHCH1 Fc杵VH (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
331 | (9B11) VHCH1 Fc臼VL (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
SEQ ID NO: | 描述 | 序列 |
259 | (12B3) VLCL-輕鏈 (核苷酸序列) | 參看表70 |
332 | (12B3) VHCH1 Fc杵VL (4B9) (HC 1之核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTGGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
333 | (12B3) VHCH1 Fc臼VH (4B9) (HC2之核苷酸序列) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTGGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTGAATTCCGTTTCTACGCTGACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
261 | (12B3) VLCL-輕鏈 | 參看表70 |
334 | (12B3) VHCH1 Fc杵VL (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
335 | (12B3) VHCH1 Fc臼VH (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSEFRFYADFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
263 | (25G7) VLCL-輕鏈 (核苷酸序列) | 參看表70 |
336 | (25G7) VHCH1 Fc杵VL (4B9) (核苷酸序列,重鏈1) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGTGACGACCCGTGGCCGCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
337 | (25G7) VHCH1 Fc臼VH (4B9) (核苷酸序列,重鏈2) | GAGGTGCAATTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCCGGATTCACCTTTAGCAGTTATGCCATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGCTATTAGTGGTAGTGGTGGTAGCACATACTACGCAGACTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAGATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGCGTGACGACCCGTGGCCGCCGTTCGACTACTGGGGCCAAGGAACCCTGGTCACCGTCTCGAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
265 | (25G7) VLCL-輕鏈 | 參看表70 |
338 | (25G7) VHCH1 Fc杵VL (4B9) (重鏈1) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
339 | (25G7) VHCH1 Fc臼VH (4B9) (重鏈2) | EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDDPWPPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
267 | (11D5) VLCL-輕鏈 (核苷酸序列) | 參看表70 |
340 | (11D5) VHCH1 Fc杵VL (4B9) (核苷酸序列,重鏈1) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTACTCTGATCTACGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
341 | (11D5) VHCH1 Fc臼VH (4B9) (核苷酸序列,重鏈2) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTACTCTGATCTACGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
269 | (11D5) VLCL-輕鏈 | 參看表70 |
342 | (11D5) VHCH1 Fc杵VL (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
343 | (11D5) VHCH1 Fc臼VH (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSTLIYGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
271 | (9B11) VLCL-輕鏈 (核苷酸序列) | 參看表70 |
344 | (9B11) VHCH1 Fc杵VL (4B9) (核苷酸序列,重鏈1) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCCTGCAGAGATGAGCTGACCAAGAACCAGGTGTCCCTGTGGTGTCTGGTCAAGGGCTTCTACCCCAGCGATATCGCCGTGGAGTGGGAGAGCAACGGCCAGCCTGAGAACAACTACAAGACCACCCCCCCTGTGCTGGACAGCGACGGCAGCTTCTTCCTGTACTCCAAACTGACCGTGGACAAGAGCCGGTGGCAGCAGGGCAACGTGTTCAGCTGCAGCGTGATGCACGAGGCCCTGCACAACCACTACACCCAGAAGTCCCTGAGCCTGAGCCCCGGCGGAGGCGGCGGAAGCGGAGGAGGAGGATCTGGGGGCGGAGGTTCCGGAGGCGGTGGATCTGAGATCGTGCTGACCCAGTCTCCCGGCACCCTGTCTCTGAGCCCTGGCGAGAGAGCCACCCTGTCCTGCAGAGCCTCCCAGTCCGTGACCTCCTCCTACCTCGCCTGGTATCAGCAGAAGCCCGGCCAGGCCCCTCGGCTGCTGATCAACGTGGGCAGTCGGAGAGCCACCGGCATCCCTGACCGGTTCTCCGGCTCTGGCTCCGGCACCGACTTCACCCTGACCATCTCCCGGCTGGAACCCGAGGACTTCGCCGTGTACTACTGCCAGCAGGGCATCATGCTGCCCCCCACCTTTGGCCAGGGCACCAAGGTGGAAATCAAG |
345 | (9B11) VHCH1 Fc臼VH (4B9) (核苷酸序列,重鏈2) | CAGGTGCAATTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGTCCTCGGTGAAGGTCTCCTGCAAGGCCTCCGGAGGCACATTCAGCAGCTACGCTATAAGCTGGGTGCGACAGGCCCCTGGACAAGGGCTCGAGTGGATGGGAGGGATCATCCCTATCTTTGGTACAGCAAACTACGCACAGAAGTTCCAGGGCAGGGTCACCATTACTGCAGACAAATCCACGAGCACAGCCTACATGGAGCTGAGCAGCCTGAGATCTGAGGACACCGCCGTGTATTACTGTGCGAGATCTTCTGGTGCTTACCCGGGTTACTTCGACTACTGGGGCCAAGGGACCACCGTGACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAAAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCTGCAGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCCTCGGCGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTGCACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTCTCGTGCGCAGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCGTGAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGAGGCGGCGGAAGCGGAGGAGGAGGATCCGGCGGCGGAGGTTCCGGAGGCGGAGGATCCGAGGTGCAGCTGCTCGAAAGCGGCGGAGGACTGGTGCAGCCTGGCGGCAGCCTGAGACTGTCTTGCGCCGCCAGCGGCTTCACCTTCAGCAGCTACGCCATGAGCTGGGTCCGCCAGGCCCCTGGCAAGGGACTGGAATGGGTGTCCGCCATCATCGGCTCTGGCGCCAGCACCTACTACGCCGACAGCGTGAAGGGCCGGTTCACCATCAGCCGGGACAACAGCAAGAACACCCTGTACCTGCAGATGAACAGCCTGCGGGCCGAGGACACCGCCGTGTACTACTGCGCCAAGGGATGGTTCGGCGGCTTCAACTACTGGGGACAGGGCACCCTGGTCACCGTGTCCAGC |
273 | (9B11) VLCL-輕鏈 | 參看表70 |
346 | (9B11) VHCH1 Fc杵VL (4B9) (重鏈1) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVTSSYLAWYQQKPGQAPRLLINVGSRRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQGIMLPPTFGQGTKVEIK |
347 | (9B11) VHCH1 Fc臼VH (4B9) (重鏈2) | QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGGIIPIFGTANYAQKFQGRVTITADKSTSTAYMELSSLRSEDTAVYYCARSSGAYPGYFDYWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALGAPIEKTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAIIGSGASTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKGWFGGFNYWGQGTLVTVSS |
純系 | 產量 [mg/l] | 單體 [%] | CE-SDS ( 非紅色) |
2+1 25G7/FAP ( 臼-VH) | 25.6 | 96.7 | 95.4 |
2+1 11D5/FAP ( 臼-VH) | 6.3 | 97 | 89.2 |
純系 | NIH/3T3-huFAP純系19細胞之EC 50[nM] | WM-266-4之EC 50[nM] |
4-1BB(25G7)/ FAP(4B9) 2+1 | 0.1 | 0.3 |
4-1BB(11D5)/ FAP(4B9) 2+1 | 0.3 | 0.4 |
圖 1A展示Fc連接之TNF受體抗原的單體形式,其用於製備TNF受體抗體。圖1B展示二聚人類TNF受體抗原Fc融合分子,其具有用於在TNF配位體存在下測試TNF受體抗體結合的C端Ha標籤(配位體阻斷特性)。圖1C中展示實例2.4中所述的實驗的示意性設置。
圖 2展示抗OX40抗體與經活化之人類CD4
+及CD8
+T細胞的結合。OX40不表現於休眠人類PBMC上(圖2A及2C)。人類PBMC OX40活化後在CD4
+及CD8
+T細胞上上調(圖2B及2D)。人類CD8
+T細胞上之OX40表現低於CD
4 +T細胞上的表現。所述純系與OX40陽性細胞之結合強度(EC
50值以及信號強度)有變化。以用作二次偵測抗體之FITC標記之抗人類IgG Fcγ特異性山羊IgG F(ab')2片段之螢光強度之中值(MFI)形式展示結合。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI來校正基線。x軸展示抗體構築體之濃度。全部OX40純系結合至經活化的OX40表現人類CD4
+T細胞,且以較低程度結合至經活化之人類CD8
+T細胞。
圖 3展示抗-OX40抗體與經活化小鼠CD4
+及CD8
+T細胞的結合。在休眠小鼠脾細胞上未偵測到OX40 (圖3A及3C)。活化後,CD4
+及CD8
+T細胞上的OX40上調(圖3B及3D)。藉由用從6-8週大雌性C57BL/6小鼠獲得的機械均質化脾臟的ACK裂解緩衝液的溶血作用分離小鼠脾細胞。使用FACS分析,用結合至FITC的山羊抗人類IgG Fc特異性二次抗體偵測抗OX40抗體與細胞表面蛋白質的結合。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI來校正基線。x軸展示抗體構築體之濃度。僅純系20B7結合至經活化之OX40表現小鼠CD4及CD8 T細胞,但不結合至休眠T細胞。
圖 4展示食蟹獼猴經活化CD4
+及CD8
+T細胞上抗OX40抗體的結合。所述純系與OX40陽性經活化食蟹獼猴CD4
+T細胞之結合強度(EC
50值以及信號強度)有變化(圖4A)。在此條件下,經活化CD8
+T細胞上的OX40表現較低且幾乎未發現所選純系的任何結合(圖4B)。使用FACS分析,用結合至FITC的山羊抗人類IgG Fc特異性二次抗體偵測抗OX40抗體與細胞表面蛋白質的結合。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI來校正基線。x軸展示抗體構築體之濃度。全部OX40純系結合至經活化之OX40表現食蟹獼猴CD4
+T細胞,且以較低程度結合至經活化食蟹獼猴CD8
+T細胞。
圖 5展示缺乏結合於OX40陰性腫瘤細胞。所述純系不顯示結合於OX40陰性U-78 MG(圖5A)及WM266-4腫瘤細胞(圖5B)。以用作二次偵測抗體之FITC標記之抗人類IgG Fcγ特異性山羊IgG F(ab')2片段之螢光強度之中值(MFI)形式展示結合。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI來校正基線。x軸展示抗體構築體之濃度。IgG型式的全部純系不結合至OX40陰性腫瘤細胞。結合對經活化白細胞上的OX40具有特異性。
圖 6A 至 6F展示如利用表面電漿子共振所量測的抗-Ox40抗體8H9 (圖6A)、20B7 (圖6B)、1G4 (圖6C)、49B4 (圖6D)、CLC-563 (圖6E)及CLC-564 (圖6F)與預先形成的複合物huOx40配位體/huOx40-Fc之間的相互作用。
圖 7展示本發明之抗人類OX40抗體對表現人類OX40及報導基因NF-kB螢光素酶之HeLa細胞的作用。所示為在具有(圖7B)或不具有(圖7B)二次抗體交聯的情況下,P329GLALA huIgG1型式中報導體細胞株中NF-kB信號傳導路徑經多種抗OX40結合子的活化。在抗OX40構築體存在下,在指定濃度之1:2比率的w/或w/o交聯二次聚-純系抗huIgG1 Fcγ-特異性山羊IgG F(ab)2片段下,培養報導體細胞6小時。藉由在0.5秒期間所量測之所釋放之光之單位(URL)與測試抗-Ox40構築體之濃度(nM)之關係的墨點分析表徵活性。由於螢光素酶介導之螢光素氧化成氧化螢光素而發出URL。當觸發人類OX40
+報導體細胞株中之OX40軸時,全部純系均能夠誘發NFκB活化。全部純系因此具有促效性且以劑量依賴性方式活化。藉由二次Fc部分特異性Ab交聯有力增加這一促效作用。
圖 8A - 8F示出了預活化人類CD4 T細胞中抗人類OX40抗體的生物活性。與培養板固定之抗Ox40結合子(huIgG1 P329GLALA型式)共刺激促進次佳再刺激人類CD4 T細胞的細胞增殖及成熟且誘發加強之活化表型。PHA-L預先活化之CFSE標記之人類CD4 T細胞在培養板上培養四天,該等培養板用小鼠IgG Fcγ特異性抗體、人類IgG Fcγ特異性抗體(皆2 μg/mL)、小鼠抗人類CD3抗體(純系OKT3,[3 ng/mL])及經滴定之抗Ox40結合子(huIgG1 P329GLALA型式)預先塗佈。所示為第4天時事件計數(圖8A)、增殖(CFSE-低)細胞百分比(圖8B)、效應子T細胞(CD127低/CD45RA低)百分比(圖8C)及CD62L低百分比(圖8D)、OX40陽性(圖8F)或Tim-3陽性細胞(圖8E)。減去僅含有培養板固定之抗人類CD3的樣品的基線值。因此,此處展示OX40刺激之增強作用,而非次佳抗CD3刺激之作用本身。當塗佈至培養板時,全部純系均能夠支持OX40陽性預先活化CD4 T細胞中的次佳TCR刺激。細胞存活的更好且增殖的更多。在腫瘤微環境中,此可導致T細胞的抗腫瘤活性增加。
圖 9概述作為各別純系的促效能力之標記物的EC50值(針對全部生物標記物)(自圖8中所示的曲線計算之值)。效能自左向右增加。第4天的事件計數、增殖(CFSE-低)細胞百分比及CD62L低、CD45RA低或Tim-3陽性細胞之百分比對比於抗Ox40抗體濃度繪製,且EC
50值在Prism4 (GraphPad Software, USA)中使用內置S形劑量反應行情計算。
圖 10A - 10F顯示抗人類OX40抗體在溶液形式的預先活化之人類CD4 T細胞中的生物活性。在無培養板固定的抗Ox40結合子(hu IgG1 P329GLALA型式)存在的情況下,偵測不到對細胞增殖、次佳再刺激之人類CD4 T細胞的成熟或活化狀態的作用。PHA-L預先活化之CFSE標記之人類CD4 T細胞在用小鼠IgG Fcγ特異性抗體及小鼠抗人類CD3抗體(純系OKT3,[3 ng/mL])預先塗佈的培養板上培養四天。經滴定之抗Ox40結合子(hu IgG1 P329GLALA型式)添加至培養基且在實驗過程中以溶液形式存在。所示為第4天時的事件計數(圖10A)、增殖(CFSE-低)細胞百分比(圖10B)、效應子T細胞(CD127低CD45RA低)百分比(圖10C)及CD62L低百分比(圖10E)、OX40陽性(圖10F)或Tim-3陽性細胞(圖10E)。減去僅含有培養板固定之抗人類CD3的樣品的基線值。因此,此處展示OX40刺激之增強作用,而非次佳抗CD3刺激之作用本身。在無強交聯(溶液形式之P329GLALA型式)存在的情況下不存在提高之TCR刺激。交聯因此對打算對T細胞具有促效性的二價aOx40型式是必需的。此交聯將由目標型式的腫瘤或腫瘤-基質細胞的細胞表面上表現之FAP提供。
圖 11展示不同抗OX40純系的結合強度與促效能力之間的相關性。抗Ox40純系(huIgG1 P329GLALA型式)在經活化CD4 T細胞上的結合如實例2.1.2中所述進行。根據使用純系8H9(huIgG1 P329GLALA型式)獲得的值對平穩值進行標準化。抗Ox40純系(huIgG1 P329GLALA型式)的生物活性測試如實例3.2中所述進行且PD-1表現的平穩值根據針對純系8H9 (huIgG1 P329GLALA型式)獲得的值標準化。標準化結合針對標準化生物活性繪圖,以測試結合強度與促效能力之間的相關性。多數純系存在直接相關性(顯示線性回歸,p值0.96;斜率0.91)。然而,兩個展現強得多的生物活性的純系(49B4,1G4)則可自其結合強度預測。縱使結合能力低但展現出乎意料地高促效效能的此純系子組對於本發明的雙特異性抗原結合分子尤其重要。
在
圖 12A中,展現包含兩個結合於OX40之Fab片段及兩個結合於FAP之交叉Fab片段的本發明的例示性雙特異性二價抗原結合分子(2+2型式)的示意性流程。
圖 12B展現包含一個結合於OX40之Fab片段及一個結合於FAP之交叉Fab片段的本發明的例示性雙特異性單價抗原結合分子(1+1型式)的示意性流程。在
圖 12C中,展現用於顯示同時結合於固定人類OX40或人類4-1BB及人類FAP的SPR實驗之設置。在
圖 12D中展現例示性雙特異性抗原結合分子之示意圖,其對於結合於OX40為二價且對於結合於FAP為單價。其包含兩個結合於OX40之Fab片段及結合於FAP之VH及VL結構域。黑點象徵重鏈中的杵-臼修飾。在
圖 12E中,展現如實例5.4.1中所述的表明結合於FAP之SPR實驗的設置。
圖 12F展示如何量測同時結合於固定人類OX40及人類FAP(實例5.4.1)。
圖 13A - 13D展現雙特異性二價2+2構築體(分析物1)同時結合於固定人類OX40及人類FAP(分析物2)的SPR圖。在
圖 13E - 13H中,展現雙特異性單價1+1構築體(分析物1)同時結合於固定人類OX40及人類FAP(分析物2)。在
圖 13I - 13K中,展現雙特異性2+1構築體(分析物1)同時結合於固定人類OX40及人類FAP(分析物2)。
圖 13L展現基於細胞之FRET分析法(TagLite)中雙特異性2+1構築體與hu OX40之結合(實例5.4.2)。資料顯示2+1構築體中的兩個抗OX40 Fab結構域以與常見IgG抗體相當之方式結合於hu OX40。
圖 14A - 14D及
14E - 14H分別展現靶向單價或二價型式之FAP中所選抗-OX40結合子(純系8H9,1G4)與休眠及活化人類PBMC的結合。與OX40陽性T細胞之結合特徵(圖14B及14D)對於習知二價hu IgG型式(空心正方形)及FAP靶向二價型式(實心正方形)中的純系是相當的。FAP靶向單價型式的同一純系(實心三角形)之結合由於損失親合力結合而明顯較弱。在無人類Ox40表現細胞存在的情況下,觀測不到結合(休眠細胞,左部曲線)。以用作二次偵測抗體之FITC標記之抗人類IgG Fcγ特異性山羊IgG F(ab')2片段之螢光強度之中值(MFI)形式展示結合。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI校正基線值(參看實例4.3.2.1)。DP88 huIgG1 P329G LALA為用作對照之同型抗體。x軸展示抗體構築體之濃度。在
圖 14A - 14D中,可見純系1G4結合於經活化OX40表現人類CD4 T細胞,及以較低程度結合於經活化人類CD8 T細胞。二價構築體比單價構築體結合得更牢固。構築體不結合於OX40陰性休眠T細胞。
圖 14E - 14H展示純系8H9結合於經活化OX40表現人類CD4 T細胞,及以較低程度結合於經活化人類CD8 T細胞。二價構築體比單價構築體結合得更牢固。構築體不結合於OX40陰性休眠T細胞。在
圖 14I - 14L中,亦展示二價FAP靶向OX40構築體相較於單價抗體型式中之各別純系對OX40陽性細胞顯示較強結合特徵。
圖 14M - 14P展示獨立於第二結合部分以類似強度結合於OX40陽性T細胞之不同2+1構築體。
圖 15A 及 15B展示FAP靶向單價或二價型式之所選抗-OX40結合子(純系8H9、1G4)與FAP陽性腫瘤細胞之結合。轉基因修飾之小鼠胚胎纖維母細胞NIH/3T3-huFAP純系39或WM266-4細胞表現高含量之人類纖維母細胞活化蛋白(huFAP)。僅FAP靶向單價及二價抗Ox40構築體(實心正方形及三角形)但並非人類IgG1 P329GLALA型式(空心正方形)中之相同純系分別結合於NIH/3T3-huFAP純系39細胞(圖15A)及Wm266-4細胞(圖15B)。以用作二次偵測抗體之異硫氰酸螢光素(FITC)標記之抗人類IgG Fcγ特異性山羊IgG F(ab')2片段之螢光強度之中值(MFI)形式展示結合。藉由流動式細胞量測術量測MFI。x軸展示抗體構築體之濃度。二價FAP構築體比單價構築體結合得更牢固。與表現人類FAP之腫瘤細胞之結合亦展示於
圖 15C 、 15D 、 15E 及 15F中(更多細節參看實例4.5.4.2)。
圖 16A 至 16G展示在有或無高度交聯存在下,單價或二價FAP靶向型式中所選結合子(8H9、1G4)對NFκB的活化。所示為報導體細胞中NF-kB信號傳導路徑經單價(實心三角形)或二價(實心正方形) FAP靶向型式或以非靶向hu IgG P329GLALA抗體(空心正方形)形式的所選結合子1G4 (圖16A-16D)及8H9 (圖16E-16G)之活化 。由抗-hu IgG Fcg特異性二次抗體(一次抗體比二次抗體之比率1:2)或經FAP表現NIH/3T3-huFAP純系39及WM266-4腫瘤細胞(FAP
+腫瘤細胞比報導體細胞之比率2:1)提供高度交聯。藉由在0.5秒期間量測之所釋放之光之單元(URL)與測試化合物之濃度(nM)之關係的墨點分析來表徵NF-κB介導之螢光素酶活性。由於螢光素酶介導之螢光素氧化成氧化螢光素而發出URL。值為藉由減去空白對照之URL校正之基線值。在圖16A中,可見全部含有純系1G4之構築體能夠誘發Ox40
+HeLa報導體細胞中之NFκB活化。二次抗IgG Fcγ特異性抗體之交聯大大增加NFκB活化。然而,添加FAP陽性細胞(NIH或WM266-4)僅增加FAP靶向分子之促效可能,但不增加P329GLALA IgG型式之促效可能。二價構築體明顯比單價構築體表現得好。在
圖 16E - 16G中,展現全部含有純系8H9之構築體能夠誘發OX40
+HeLa報導體細胞中之NFκB活化。二次抗IgG Fcγ特異性抗體之交聯大大增加NFκB活化。然而,添加FAP陽性細胞(NIH)僅提高FAP靶向分子之促效可能,但不提高P329GLALA IgG型式之促效可能。二價構築體比單價構築體表現得略好。在
圖 16H 、 16I 及 16J中,亦可發現單價結合於OX40 (1+1)之構築體比二價結合於OX40 (2+1及2+2構築體之構築體低效。其他資料於
圖 16K 、 16L 及 16M及實例5.1中給出。
預先活化之CD4 T細胞經培養板固定之FAP靶向單價及二價抗OX40 (1G4及8H9)構築體的次佳TCR再刺激的補救展示於
圖 17A 及 17B 以及圖 18A - 18D中。與培養板固定之抗Ox40結合子(huIgG1 P329GLALA型式)共刺激促進次佳再刺激人類CD4 T細胞的細胞增殖及成熟且誘發加強之活化表型。PHA-L預先活化之CFSE標記之人類CD4 T細胞在培養板上培養四天,該等培養板用小鼠IgG Fcγ特異性抗體、人類IgG Fcγ特異性抗體(皆2 μg/mL)、小鼠抗人類CD3抗體(純系OKT3,[3 ng/mL])及經滴定之抗Ox40結合子(huIgG1 P329GLALA型式)預先塗佈。所示為第4天時的事件計數,增殖(CFSE-低)細胞百分比、效應子T細胞(CD127低CD45RA低)百分比及CD62L低、OX40陽性或Tim-3陽性細胞之百分比。減去僅含有培養板固定之抗人類CD3的樣品的基線值。因此,此處展示OX40刺激之增強作用,而非次佳抗CD3刺激之作用本身。在圖17中,可見全部含有純系8H9之構築體能夠補救塗佈於培養板時預先活化之Ox40
+CD4 T細胞的次佳TCR刺激。細胞展現更大程度活化(Tim3
+FSC
+)表型。二價構築體比單價構築體表現得略好。在圖18中,可觀測到全部含有純系1G4之構築體能夠補救塗佈於培養板時預先活化之Ox40
+CD4 T細胞的次佳TCR刺激。細胞進一步增殖且呈現經活化(Tim3
+Ox40
+)表型。二價構築體表現得比單價構築體好。當塗佈於培養板時,二價構築體皆表現得相當。
在
圖 19中,展示自用培養板固定之FAP靶向單價及二價抗OX40(純系1G4)構築體補救次佳TCR刺激計算的EC
50值。第4天的增殖(CFSE-低)細胞百分比及CD127L低、Tim-3陽性及OX40陽性細胞之百分比對比於抗OX40抗體濃度繪製,且作為促效強度之量度的EC
50值在Prism4 (GraphPad Software, USA)中使用內置S形劑量反應行情計算。全部含有純系1G4之構築體能夠補救塗佈於培養板時預先活化之Ox40
+CD4 T細胞的次佳TCR刺激。然而,二價(2+2)構築體比單價(1+1)構築體表現得好。
圖20A-20H涉及次佳TCR觸發之休眠人類PBMC及細胞表面FAP高度交聯的OX40介導之共刺激。僅含有FAP結合部分之構築體能夠補救由FAP陽性細胞(NIH)提供交聯時預先活化之Ox40
+CD4 T細胞的次佳TCR刺激。展示事件計數(圖20B及20D)、低增殖(CFSE-高)細胞之百分比(圖20A及20C)或CD62L之MFI (圖20F及20H)、CD127 (圖20E)或顆粒酶B重要CD4
+(圖20G)及CD8
+T細胞。減去僅含有抗人類CD3 (純系V9,huIgG1)、休眠人類PBMC及NIH/3T3-huFAP純系39之樣品的基線值。因此,此處可見OX40共刺激之增強作用,而非次佳抗CD3刺激之作用本身。細胞存活得更好,增殖得更多且展現更強活化(CD62L及CD127低)表型。靶向二價構築體比單價構築體表現得僅略好。非靶向huIgG1P329GLALA中的純系8H9在無進一步交聯存在下不能補救次佳TCR刺激。在FAP陽性腫瘤微環境中,此可能導致增加的T細胞抗腫瘤活性,由此避免全身性OX40活化。
圖 21A - 21C涉及使用表面固定之FAP靶向單價及二價抗OX40 (1G4)構築體活化休眠人類CD4細胞。用非靶向抗Ox40 (1G4) huIgG1共刺激不會補救次佳TCR刺激之CD4及CD8 (資料未示出) T細胞。FAP靶向單價及二價抗Ox40構築體與本發明NIH/3T3-huFAP純系39細胞之高度交聯大大促進人類CD4細胞之增殖、存活(資料未示出)且誘導加強活化表型。所示為CD4 T細胞上CD25表現之MFI及CD25
+CD4 T細胞的百分比。減去僅含有抗人類CD3 (純系V9,huIgG1)、休眠人類PBMC及NIH/3T3-huFAP純系39之樣品的基線值。化合物之促效作用使用GraphPad Prism之內置函數根據曲線下面積定量且針對三種不同抗Ox40 (1G4)構築體顯示。靶向二價(2+2)構築體比單價(1+1)構築體表現得好。
第二實驗中獲得之資料分別顯示於
圖 21D - 21H 及 21I - 21M中。單價抗OX40構築體(1+1;實心三角形)補救TCR刺激的能力低於二價抗OX40靶向構築體(半實心圓,實心正方形)。FAP與2+2構築體之二價結合相較於FAP與2+1構築體之單價結合已經能夠在較低濃度下補救次佳TCR刺激。在2+1型式中,高親和力FAP結合純系4B9明顯優於低親和力純系28H1 (
圖 21I - 21M)。此表明觀測到的生物活性的EC
50值由與FAP之結合(2+2 > 2+1 (4B9) > 2+1 (28H1))驅動。在
圖 21N中,各構築體之促效能力根據曲線下面積針對分析之標記物定量且針對彼此繪製。FAP (28H1) (2+2)構築體觀測到的生物活性最佳,隨後是FAP (4B9) (2+1)構築體,接著是FAP (28H1) (2+1)構築體。
在
圖 22A 及 22B中,已知含有人類IgG1 Fc區之抗Ox40抗體可誘發OX40陽性細胞溶解。PkH26標記之HeLa_hOx40_NFκB_Luc1及新鮮分離之NK細胞在E比T比率3:1下,在OX40抗體(人類IgG1及人類IgG1 P329GLALA)存在下共培養24小時。4小時後使用細胞毒性偵測試劑盒-LDH(Roche, 目錄號11644793001)分析LDH含量。24小時候,細胞用Dapi染色且藉由流動式細胞量測術分析。Dapi陽性死細胞之百分比用於計算特異性溶解。人類IgG型式中之抗OX40結合子結合於NK細胞上的Fc受體且誘發OX40陽性目標細胞之ADCC。實際上使用hu IgG1 P329GLALA型式防止OX40陽性細胞(例如最近活化之T細胞)的ADCC。
圖 23A - 23D展示與轉移至huIgG1 P329G LALA型式的四種抗人類4-1BB特異性純系(實心菱形:純系25G7,實心正方形:純系12B3,實心星形:純系11D5,向上三角形:純系9B11)及轉移至huIgG1 P329G LALA型式的一種抗小鼠4-1BB特異性純系20G2 (向下三角形)的休眠及活化人類T細胞的結合。使用非4-1BB特異性純系DP47 huIgG1 P329G LALA抗體作為陰性對照(空心灰色圓)。上圖展示與休眠CD4
+T細胞(圖23A)及經活化之CD4
+T細胞(圖23B)的結合,而下圖展示與休眠CD8
+T細胞(圖23C)及經活化之CD8
+T細胞(圖23D)的結合。結合之特徵為用作二次偵測抗體之FITC標記或PE標記之抗人類IgG Fcγ特異性山羊IgG F(ab`)
2片段的螢光中位值(MFI)對比所測試之一次抗4-1BB結合huIgG1 P329G LALA抗體之濃度(nM)繪圖。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI校正基線值(無一次抗體)。
圖 24A - 24D展示與4-1BB表現小鼠T細胞之結合。所示為與四種抗人類4-1BB結合huIgG1 P329G LALA抗體純系(實心菱形:純系25G7,實心正方形:純系12B3、實心星形:純系11D5、向上三角形:純系9B11)及一種抗小鼠4-1BB結合huIgG1 P329G LALA抗體純系20G2 (向下三角形)的休眠及活化小鼠T細胞的結合。使用非4-1BB結合DP47 huIgG1 P329G LALA抗體作為陰性對照(空心灰色圓)。上圖展示與休眠小鼠CD4
+T細胞(圖24A)及經活化之CD4
+T細胞(圖24B)的結合,而下圖展示與休眠小鼠CD8
+T細胞(圖24C)及經活化之CD8
+T細胞(圖24D)的結合。結合之特徵為用作二次偵測抗體之FITC標記之抗人類IgG Fcγ特異性山羊IgG F(ab`)
2片段的MFI對比所測試之一次抗4-1BB結合huIgG1 P329G LALA抗體之濃度(nM)繪圖。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI校正基線值(無一次抗體)。
圖 25A - 25D展示小鼠IgG與4-1BB表現小鼠T細胞之結合。所示為與轉移至小鼠IgG1 DAPG及小鼠IgG1野生型(wt)型式的抗小鼠4-1BB結合純系20G2的休眠及活化小鼠T細胞的結合。使用市售非4-1BB結合小鼠IgG1 wt同型對照(空心灰色圓,BioLegend, 目錄號400153)作為陰性對照。上圖展示與休眠CD4
+T細胞(圖25A)及經活化之CD4
+T細胞(圖25B)的結合,而下圖展示與休眠CD8
+T細胞(圖25C)及經活化之CD8
+T細胞(圖25D)的結合。結合之特徵為用作二次偵測抗體之FITC標記之抗小鼠IgG Fcγ特異性山羊IgG F(ab`)
2片段的螢光強度中位值(MFI)對比所測試之一次抗4-1BB結合moIgG抗體之濃度(nM)繪圖。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI校正基線值(無一次抗體)。
圖 26A 及 26B展示與4-1BB表現食蟹獼猴T細胞之結合。所示為與四種抗人類4-1BB結合huIgG1 P329G LALA抗體純系(實心菱形:純系25G7、實心正方形:純系12B3、實心星形:純系11D5、向上三角形:純系9B11)之經活化食蟹獼猴T細胞之結合。使用非4-1BB結合DP47 huIgG1 P329G LALA抗體作為陰性對照(空心灰色圓)。所示分別為與經活化CD4
+T細胞(圖26A)及經活化CD8
+T細胞(圖26B)的結合。結合之特徵為用作二次偵測抗體之FITC標記之抗人類IgG Fcγ特異性山羊IgG F(ab`)
2片段的螢光強度中位值(MFI)對比所測試之一次抗4-1BB結合huIgG1 P329G LALA抗體之濃度(nM)繪圖。藉由流動式細胞量測術量測MFI且藉由減去空白對照之MFI校正基線值(無一次抗體)。
圖 27A - 27E涉及如表面電漿子共振所測定的本發明之抗4-1BB抗體的配位體結合特性。展示人類抗4-1BB IgG 25G7、11D5、9B11及12B3與預先形成的複合物hu4-1BB配位體/hu4-1BB之間的相互作用以及小鼠抗4-1BB純系20G2與預先形成的複合物mu4-1BB配位體/mu4-1BB之間的相互作用。
圖 28A - 28C 及 28D - 28F涉及競爭結合實驗。圖28A-28C展現抗4-1BB IgG純系12B3、11D5及25G7與預先形成的純系9B11及hu4-1BB複合物之間的相互作用。圖28D-28F展現抗4-1BB IgG純系12B3、9B11及25G7與預先形成的純系11D5及hu4-1BB複合物之間的相互作用。可以推斷抗4-1BB純系12B3、11D5及9B11與25G7共用不同空間抗原決定基,因為兩個抗體可同時結合於人類4-1BB。
圖 29A - 29D展示雜交4-1BB Fc(kih)變異體與抗4-1BB抗體之結合,即hu4-1BBD1/mu4-1BBD2-Fc(kih)及mu4-1BBD1/hu4-1BBD2-Fc(kih)變異體與抗4-1BB抗體之結合。帶下劃線的是抗體識別之4-1BB結構域。在
圖 30A - 30D中,展示抗人類4-1BB抗體11D5、12B3、25G7及9B11與人類4-1BB結構域1之結合。抗人類4-1BB抗體11D5、12B3及9B11結合含有人類結構域1之4-1BB構築體。
圖 31A - 31D展現不同抗人類4-1BB純系之活體外功能特性。預先活化之人類CD8
+T細胞在無抗人類CD3抗體存在下(圖31A及31C)或在次佳濃度之表面固定抗人類CD3抗體存在下(圖31B及31D)用不同濃度之表面固定抗人類4-1BB特異性huIgG1 P329G LALA抗體活化。所示為總CD8
+T細胞群體中IFNγ
+(A及B)及TNFα
+(C及D) CD8
+T細胞之頻率對比表面固定4-1BB結合huIgG1 P329G LALA之濃度(pM)。在CD3刺激存在下,4-1BB共刺激可以濃度依賴性方式增加IFNγ (圖31B)及TNFα (圖31D)分泌。在無CD3刺激存在下,4-1BB之活化對IFNγ (圖31A)及TNFα (圖31C)分泌無作用。
圖 32A 及 32B展示抗小鼠4-1BB純系20G2之活體外功能特性。在無0.5 μg/mL抗小鼠IgG1 CD3倉鼠IgG (純系145-2C11)及溶液中不同濃度之抗小鼠4-1BB抗體(實心黑色菱形:小鼠IgG,空心黑色菱形:小鼠IgG DAPG)或適合同型對照(實心灰色圓:小鼠IgG1、空心灰色圓:小鼠IgG1 DAPG)存在下培育小鼠脾細胞。濃縮在x軸上表示,以nM為單位。僅當抗小鼠4-1BB純系20G2小鼠IgG1 (黑色菱形)可經FcR表現細胞交聯時,顆粒酶B (圖32A)及脫中胚蛋白(圖32B)之活化可以濃度依賴性方式增加。
圖 33展示抗小鼠4-1BB純系20G2之活體內功能特性。所示為每組三隻小鼠之結果。用抗小鼠4-1BB純系處理後,20G2小鼠IgG1 (灰色條) CD8
+T細胞以總數(a)在肝臟中積聚。CD8
+T細胞上的另一增殖標記物Ki67以頻率(b)及總數(c)上調。其亦藉由以頻率(d)及總數(e)上調CD8
+T細胞上的4-1BB (CD137)來誘發正回饋迴路。在第三次注射之後1天發現最強作用。若小鼠用抗小鼠4-1BB純系20G2小鼠IgG1 DAPG處理(黑色條),則防止抗體交聯且不會發生4-1BB活化。因此,肝臟中之CD8
+T細胞的數目及表型與PBS處理之小鼠(白色條)類似。
在
圖 34A中,展現包含兩個結合於4-1BB之Fab片段及兩個結合於FAP之交叉Fab片段的本發明的例示性雙特異性二價抗原結合分子(2+2型式)的示意性流程。在
圖 34B中,展現用於顯示同時結合於固定人類4-1BB及人類FAP的SPR實驗之設置。
圖 34C展示包含一個結合於4-1BB之Fab片段及一個結合於FAP之交叉Fab片段的本發明的例示性雙特異性單價抗原結合分子(1+1型式)的示意性流程。
雙特異性二價抗4-1BB/抗FAP構築體之同時結合顯示於
圖 35A - 35D中。雙特異性構築體用作固定人類4-1BB之分析物1且人類FAP用作分析物2。全部雙特異性構築體可同時結合人類4-1BB及人類FAP。
圖 36A 及 36B展示例示性雙特異性抗原結合分子,其為二價抗4-1BB及單價抗FAP huIgG1 P329GLALA,亦稱為2+1型式。雙特異性抗原結合分子包含兩個結合於4-1BB之Fab片段及結合於FAP之VH及VL結構域。
圖 37A - 37C涉及雙特異性2+1抗4-1BB及抗FAP構築體的同時結合。
圖 37A為分析設備之圖形符號;圖37B及37C展示2+1型式的雙特異性抗原結合分子(分析物1)與固定人類4-1BB及人類FAP的所偵測的同時結合。
圖 38A - 38F展示與人類4-1BB特異性純系11D5 (
圖 38A 及 C)、12B3 (
圖 38B 及 D)以及25G7 (
圖 38E 及 F)之休眠CD4
+(上圖)及CD8
+T細胞(下圖)的結合。結合以二次檢測抗體PE結合之抗人類IgG Fcγ-片段-特異性山羊IgG F(ab2')片段對比一次4-1BB結合抗體之濃度的螢光強度幾何平均值呈現。在全部墨點中,使用陰性對照DP47未靶向之huIgG1 P329G LALA (空心黑色圓,點劃線)。無構築體展現與休眠人類CD4
+T細胞(
圖 38A 、 B 及 E)或休眠CD8
+T細胞(
圖 38C 、 D 及 F)之特異性結合。
圖 39A - 39F展示與人類4-1BB特異性純系11D5 (
圖 39A 及 C)、12B3 (
圖 39B 及 D)以及25G7 (
圖 39E 及 F)之活化CD4
+(上圖)及CD8
+T細胞(下圖)的結合。結合以二次檢測抗體PE結合之抗人類IgG Fcγ-片段-特異性山羊IgG F(ab2')片段對比一次4-1BB結合抗體之濃度的螢光強度幾何平均值顯示。在全部墨點中,使用陰性對照DP47未靶向之huIgG1 P329G LALA (空心黑色圓,點劃線)。全部構築體主要結合於活化人類CD8
+T細胞(
圖 39 C 、 D 及 F),其展現比活化人類CD4
+T細胞高的4-1BB表現(
圖 40 A 、 B 及 E)。
圖 40概述根據結合曲線之曲線下面積(AUC)與不同純系及型式之活化人類CD8
+T細胞的結合。不同型式及所用抗FAP結合純系以曲線下之圖形符號表示,4-1BB結合純系由柱形圖案表示:DP47對照分子為白色,含有25G7之分子為黑色,若未靶向之DP47為黑白條,則純系11D5為灰色且純系12B3為白色/黑色格子。
圖 41A - 41F展示與人類FAP表現黑素瘤細胞株WM-266-4 (
圖 42 A 、 B 及 E)以及NIH/3T3-huFAP純系19細胞(
圖 42 C 、 D 及 F)的結合。結合以二次檢測抗體PE結合之抗人類IgG Fcγ-片段-特異性山羊IgG F(ab2')片段對比一次4-1BB結合抗體之濃度的螢光強度幾何平均值顯示。使用含有4-1BB結合純系11D5的構築體的結合曲線展示於圖41A及41C中,純系12B3展示於圖41B及41D且純系25G7展示於圖41E及41F中。在全部墨點中,使用陰性對照DP47未靶向之huIgG1 P329G LALA (空心黑色圓,點劃線)。僅FAP靶向型式結合於FAP表現細胞且其親本抗4-1BB huIgG1P329G LALA抗體不結合。因此,與型式無關,全部所示FAP靶向分子特徵為FAP特異性靶向特性。視型式、FAP結合純系及靶向部分而定,一些分子的FAP靶向特性比其它分子更佳。
圖 42概述與NIH/3T3-huFAP細胞之結合。所示為結合曲線的曲線下面積(AUC)。所用抗體型式以曲線下之圖形符號表示,4-1BB結合純系由彩色柱表示:DP47對照分子為白色,含有25G7之分子為黑色,若未靶向之DP47為黑白條,則純系11D5為灰色且純系12B3為白色/黑色格子。曲線展示僅FAP靶向分子而非其4-1BB結合親本huIgG1 P329G LALA或DP47靶向4-1BB (25G7)結合分子可結合於FAP表現細胞。
圖 43A - 43I展示4-1BB表現報導體細胞株HeLa-hu4-1BB-NFκB-luc中的NF-κB介導之螢光素酶活性。螢光素酶活性以釋放之光單位(URL)作為單位顯示於y軸,與培育6小時後促效人類4-1BB結合分子的添加濃度對比。在
圖 43A 、 D 及 G中,未添加FAP表現腫瘤細胞。在
圖 43B 、 E 及 H中,FAP表現人類黑素瘤細胞株WM-266-4及在
圖 43 C 、 F 及 I中,人類FAP轉染NIH/3T3細胞以比率5:1添加至報導體細胞株且培育6小時。使用含有4-1BB結合純系11D5的構築體的活化曲線展示於圖43A、43B及43C中,純系12B3展示於圖43D、43E及43F且純系25G7展示於圖43G、43H及43I中。僅FAP靶向型式在FAP表現腫瘤細胞存在下誘發螢光素酶活性。活化水準取決於純系、型式及FAP表現腫瘤細胞株。
圖 44A 及 44B概述在NIH/3T3-huFAP細胞存在下,4-1BB表現報導體細胞株HeLa-hu4-1BB-NFκB-luc中NF-κB介導之螢光素酶活性。所示為在NIH/3T3-huFAP細胞存在下,活化曲線的曲線下面積(AUC)。所用抗體型式及抗FAP純系以曲線下之圖形符號表示,不同促效4-1BB純系由不同柱模式表示:DP47對照分子為白色,含有25G7之分子為黑色,純系11D5為灰色且純系12B3為白色/黑色格子。曲線展示僅FAP靶向分子可誘發高於背景的強活化。活化水準取決於純系、FAP靶向及型式。
<![CDATA[<110> 瑞士商赫孚孟拉羅股份公司(F. Hoffmann-La Roche AG)]]> <![CDATA[<120> 對共刺激TNF受體具特異性之雙特異性抗體]]> <![CDATA[<130> TW 105131817]]> <![CDATA[<150> EP 15188095.2]]> <![CDATA[<151> 2015-10-02]]> <![CDATA[<150> EP 16170363.2]]> <![CDATA[<151> 2016-05-19]]> <![CDATA[<160> 347 ]]> <![CDATA[<170> PatentIn version 3.5]]> <![CDATA[<210> 1]]> <![CDATA[<211> 186]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 1]]> Leu His Cys Val Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys His 1 5 10 15 Glu Cys Arg Pro Gly Asn Gly Met Val Ser Arg Cys Ser Arg Ser Gln 20 25 30 Asn Thr Val Cys Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val 35 40 45 Ser Ser Lys Pro Cys Lys Pro Cys Thr Trp Cys Asn Leu Arg Ser Gly 50 55 60 Ser Glu Arg Lys Gln Leu Cys Thr Ala Thr Gln Asp Thr Val Cys Arg 65 70 75 80 Cys Arg Ala Gly Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp 85 90 95 Cys Ala Pro Cys Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala 100 105 110 Cys Lys Pro Trp Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln 115 120 125 Pro Ala Ser Asn Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro 130 135 140 Ala Thr Gln Pro Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Ile Thr 145 150 155 160 Val Gln Pro Thr Glu Ala Trp Pro Arg Thr Ser Gln Gly Pro Ser Thr 165 170 175 Arg Pro Val Glu Val Pro Gly Gly Arg Ala 180 185 <![CDATA[<210> 2]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9,49B4,1G4, 20B7) CDR-H1]]> <![CDATA[<400> 2]]> Ser Tyr Ala Ile Ser 1 5 <![CDATA[<210> 3]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563, CLC-564, 17A9) CDR-H1]]> <![CDATA[<400> 3]]> Ser Tyr Ala Met Ser 1 5 <![CDATA[<210> 4]]> <![CDATA[<211> 17]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9,49B4,1G4, 20B7) CDR-H2]]> <![CDATA[<400> 4]]> Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <![CDATA[<210> 5]]> <![CDATA[<211> 17]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563, CLC-564, 17A9) CDR-H2]]> <![CDATA[<400> 5]]> Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <![CDATA[<210> 6]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9) CDR-H3]]> <![CDATA[<400> 6]]> Glu Tyr Gly Trp Met Asp Tyr 1 5 <![CDATA[<210> 7]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(49B4) CDR-H3]]> <![CDATA[<400> 7]]> Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr 1 5 <![CDATA[<210> 8]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(1G4) CDR-H3]]> <![CDATA[<400> 8]]> Glu Tyr Gly Ser Met Asp Tyr 1 5 <![CDATA[<210> 9]]> <![CDATA[<211> 13]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(20B7) CDR-H3]]> <![CDATA[<400> 9]]> Val Asn Tyr Pro Tyr Ser Tyr Trp Gly Asp Phe Asp Tyr 1 5 10 <![CDATA[<210> 10]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563) CDR-H3]]> <![CDATA[<400> 10]]> Asp Val Gly Ala Phe Asp Tyr 1 5 <![CDATA[<210> 11]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-564) CDR-H3]]> <![CDATA[<400> 11]]> Asp Val Gly Pro Phe Asp Tyr 1 5 <![CDATA[<210> 12]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(17A9)-CDR-H3]]> <![CDATA[<400> 12]]> Val Phe Tyr Arg Gly Gly Val Ser Met Asp Tyr 1 5 10 <![CDATA[<210> 13]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9,49B4,1G4, 20B7) CDR-L1]]> <![CDATA[<400> 13]]> Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala 1 5 10 <![CDATA[<210> 14]]> <![CDATA[<211> 12]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563, CLC564) CDR-L1]]> <![CDATA[<400> 14]]> Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala 1 5 10 <![CDATA[<210> 15]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(17A9) CDR-L1]]> <![CDATA[<400> 15]]> Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser 1 5 10 <![CDATA[<210> 16]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9,49B4,1G4, 20B7) CDR-L2]]> <![CDATA[<400> 16]]> Asp Ala Ser Ser Leu Glu Ser 1 5 <![CDATA[<210> 17]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563, CLC564) CDR-L2]]> <![CDATA[<400> 17]]> Gly Ala Ser Ser Arg Ala Thr 1 5 <![CDATA[<210> 18]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(17A9) CDR-L2]]> <![CDATA[<400> 18]]> Gly Lys Asn Asn Arg Pro Ser 1 5 <![CDATA[<210> 19]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9) CDR-L3]]> <![CDATA[<400> 19]]> Gln Gln Tyr Leu Thr Tyr Ser Arg Phe Thr 1 5 10 <![CDATA[<210> 20]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(49B4) CDR-L3]]> <![CDATA[<400> 20]]> Gln Gln Tyr Ser Ser Gln Pro Tyr Thr 1 5 <![CDATA[<210> 21]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(1G4) CDR-L3]]> <![CDATA[<400> 21]]> Gln Gln Tyr Ile Ser Tyr Ser Met Leu Thr 1 5 10 <![CDATA[<210> 22]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(20B7) CDR-L3]]> <![CDATA[<400> 22]]> Gln Gln Tyr Gln Ala Phe Ser Leu Thr 1 5 <![CDATA[<210> 23]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563, CLC-164) CDR-L3]]> <![CDATA[<400> 23]]> Gln Gln Tyr Gly Ser Ser Pro Leu Thr 1 5 <![CDATA[<210> 24]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(17A9) CDR-L3]]> <![CDATA[<400> 24]]> Asn Ser Arg Val Met Pro His Asn Arg Val 1 5 10 <![CDATA[<210> 25]]> <![CDATA[<211> 116]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9) VH]]> <![CDATA[<400> 25]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Trp Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <![CDATA[<210> 26]]> <![CDATA[<211> 108]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(8H9) VL]]> <![CDATA[<400> 26]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Thr Tyr Ser Arg 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 27]]> <![CDATA[<211> 118]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(49B4) VH]]> <![CDATA[<400> 27]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser 115 <![CDATA[<210> 28]]> <![CDATA[<211> 107]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(49B4) VL]]> <![CDATA[<400> 28]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Gln Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 29]]> <![CDATA[<211> 116]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(1G4) VH]]> <![CDATA[<400> 29]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Ser Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser 115 <![CDATA[<210> 30]]> <![CDATA[<211> 108]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(1G4) VL]]> <![CDATA[<400> 30]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ile Ser Tyr Ser Met 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 31]]> <![CDATA[<211> 122]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(20B7) VH]]> <![CDATA[<400> 31]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Asn Tyr Pro Tyr Ser Tyr Trp Gly Asp Phe Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser 115 120 <![CDATA[<210> 32]]> <![CDATA[<211> 107]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(20B7) VL]]> <![CDATA[<400> 32]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gln Ala Phe Ser Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 33]]> <![CDATA[<211> 116]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563) VH]]> <![CDATA[<400> 33]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Leu Asp Val Gly Ala Phe Asp Tyr Trp Gly Gln Gly Ala Leu Val 100 105 110 Thr Val Ser Ser 115 <![CDATA[<210> 34]]> <![CDATA[<211> 108]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-563) VL]]> <![CDATA[<400> 34]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 35]]> <![CDATA[<211> 116]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-564) VH]]> <![CDATA[<400> 35]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Phe Asp Val Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <![CDATA[<210> 36]]> <![CDATA[<211> 108]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(CLC-564) VL]]> <![CDATA[<400> 36]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 37]]> <![CDATA[<211> 120]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(17A9) VH]]> <![CDATA[<400> 37]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Phe Tyr Arg Gly Gly Val Ser Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser 115 120 <![CDATA[<210> 38]]> <![CDATA[<211> 106]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> OX40(17A9) VL]]> <![CDATA[<400> 38]]> Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Val Met Pro His Asn Arg 85 90 95 Val Phe Gly Gly Gly Thr Lys Leu Thr Val 100 105 <![CDATA[<210> 39]]> <![CDATA[<211> 163]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 39]]> Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp 85 90 95 Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro 100 105 110 Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr 115 120 125 Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro 130 135 140 Gly Ala Ser Ser Val Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His 145 150 155 160 Ser Pro Gln <![CDATA[<210> 40]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3,11D5, 9B11, 20G2) CDR-H1]]> <![CDATA[<400> 40]]> Ser Tyr Ala Ile Ser 1 5 <![CDATA[<210> 41]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) CDR-H1]]> <![CDATA[<400> 41]]> Ser Tyr Ala Met Ser 1 5 <![CDATA[<210> 42]]> <![CDATA[<211> 17]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3,11D5, 9B11, 20G2) CDR-H2]]> <![CDATA[<400> 42]]> Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe Gln 1 5 10 15 Gly <![CDATA[<210> 43]]> <![CDATA[<211> 17]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) CDR-H2]]> <![CDATA[<400> 43]]> Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <![CDATA[<210> 44]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3) CDR-H3]]> <![CDATA[<400> 44]]> Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr 1 5 10 <![CDATA[<210> 45]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) CDR-H3]]> <![CDATA[<400> 45]]> Asp Asp Pro Trp Pro Pro Phe Asp Tyr 1 5 <![CDATA[<210> 46]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(11D5) CDR-H3]]> <![CDATA[<400> 46]]> Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr 1 5 10 <![CDATA[<210> 47]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(9B11) CDR-H3]]> <![CDATA[<400> 47]]> Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr 1 5 10 <![CDATA[<210> 48]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(20G2) CDR-H3]]> <![CDATA[<400> 48]]> Ser Tyr Tyr Trp Glu Ser Tyr Pro Phe Asp Tyr 1 5 10 <![CDATA[<210> 49]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3,11D5, 9B11, 20G2) CDR-L1]]> <![CDATA[<400> 49]]> Arg Ala Ser Gln Ser Ile Ser Ser Trp Leu Ala 1 5 10 <![CDATA[<210> 50]]> <![CDATA[<211> 11]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) CDR-L1]]> <![CDATA[<400> 50]]> Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala Ser 1 5 10 <![CDATA[<210> 51]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3,11D5, 9B11, 20G2) CDR-L2]]> <![CDATA[<400> 51]]> Asp Ala Ser Ser Leu Glu Ser 1 5 <![CDATA[<210> 52]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) CDR-L2]]> <![CDATA[<400> 52]]> Gly Lys Asn Asn Arg Pro Ser 1 5 <![CDATA[<210> 53]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3) CDR-L3]]> <![CDATA[<400> 53]]> Gln Gln Tyr His Ser Tyr Pro Gln Thr 1 5 <![CDATA[<210> 54]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) CDR-L3]]> <![CDATA[<400> 54]]> Asn Ser Leu Asp Arg Arg Gly Met Trp Val 1 5 10 <![CDATA[<210> 55]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(11D5) CDR-L3]]> <![CDATA[<400> 55]]> Gln Gln Leu Asn Ser Tyr Pro Gln Thr 1 5 <![CDATA[<210> 56]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(9B11) CDR-L3]]> <![CDATA[<400> 56]]> Gln Gln Val Asn Ser Tyr Pro Gln Thr 1 5 <![CDATA[<210> 57]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(20G2) CDR-L3]]> <![CDATA[<400> 57]]> Gln Gln Gln His Ser Tyr Tyr Thr 1 5 <![CDATA[<210> 58]]> <![CDATA[<211> 120]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3) VH]]> <![CDATA[<400> 58]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <![CDATA[<210> 59]]> <![CDATA[<211> 107]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(12B3) VL]]> <![CDATA[<400> 59]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr His Ser Tyr Pro Gln 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 60]]> <![CDATA[<211> 118]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) VH]]> <![CDATA[<400> 60]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser 115 <![CDATA[<210> 61]]> <![CDATA[<211> 106]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(25G7) VL]]> <![CDATA[<400> 61]]> Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Leu Asp Arg Arg Gly Met Trp 85 90 95 Val Phe Gly Gly Gly Thr Lys Leu Thr Val 100 105 <![CDATA[<210> 62]]> <![CDATA[<211> 119]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(11D5) VH]]> <![CDATA[<400> 62]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser 115 <![CDATA[<210> 63]]> <![CDATA[<211> 107]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(11D5) VL]]> <![CDATA[<400> 63]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Gln 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 64]]> <![CDATA[<211> 120]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(9B11) VH]]> <![CDATA[<400> 64]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <![CDATA[<210> 65]]> <![CDATA[<211> 107]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(9B11) VL]]> <![CDATA[<400> 65]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Val Asn Ser Tyr Pro Gln 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 66]]> <![CDATA[<211> 120]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(20G2) VH]]> <![CDATA[<400> 66]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Tyr Tyr Trp Glu Ser Tyr Pro Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser 115 120 <![CDATA[<210> 67]]> <![CDATA[<211> 106]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 4-1BB(20G2) VL]]> <![CDATA[<400> 67]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln His Ser Tyr Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 68]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) CDR-H1]]> <![CDATA[<400> 68]]> Ser His Ala Met Ser 1 5 <![CDATA[<210> 69]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) CDR-H1]]> <![CDATA[<400> 69]]> Ser Tyr Ala Met Ser 1 5 <![CDATA[<210> 70]]> <![CDATA[<211> 16]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) CDR-H2]]> <![CDATA[<400> 70]]> Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val Lys Gly 1 5 10 15 <![CDATA[<210> 71]]> <![CDATA[<211> 17]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) CDR-H2]]> <![CDATA[<400> 71]]> Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr Ala Asp Ser Val Lys 1 5 10 15 Gly <![CDATA[<210> 72]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) CDR-H3]]> <![CDATA[<400> 72]]> Gly Trp Leu Gly Asn Phe Asp Tyr 1 5 <![CDATA[<210> 73]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) CDR-H3]]> <![CDATA[<400> 73]]> Gly Trp Phe Gly Gly Phe Asn Tyr 1 5 <![CDATA[<210> 74]]> <![CDATA[<211> 12]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) CDR-L1]]> <![CDATA[<400> 74]]> Arg Ala Ser Gln Ser Val Ser Arg Ser Tyr Leu Ala 1 5 10 <![CDATA[<210> 75]]> <![CDATA[<211> 12]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) CDR-L1]]> <![CDATA[<400> 75]]> Arg Ala Ser Gln Ser Val Thr Ser Ser Tyr Leu Ala 1 5 10 <![CDATA[<210> 76]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) CDR-L2]]> <![CDATA[<400> 76]]> Gly Ala Ser Thr Arg Ala Thr 1 5 <![CDATA[<210> 77]]> <![CDATA[<211> 7]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) CDR-L2]]> <![CDATA[<400> 77]]> Val Gly Ser Arg Arg Ala Thr 1 5 <![CDATA[<210> 78]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) CDR-L3]]> <![CDATA[<400> 78]]> Gln Gln Gly Gln Val Ile Pro Pro Thr 1 5 <![CDATA[<210> 79]]> <![CDATA[<211> 9]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) CDR-L3]]> <![CDATA[<400> 79]]> Gln Gln Gly Ile Met Leu Pro Pro Thr 1 5 <![CDATA[<210> 80]]> <![CDATA[<211> 116]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) VH]]> <![CDATA[<400> 80]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser 115 <![CDATA[<210> 81]]> <![CDATA[<211> 108]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(28H1) VL]]> <![CDATA[<400> 81]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Ile Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Gln Val Ile Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 82]]> <![CDATA[<211> 117]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) VH]]> <![CDATA[<400> 82]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gln Gly Thr Leu 100 105 110 Val Thr Val Ser Ser 115 <![CDATA[<210> 83]]> <![CDATA[<211> 108]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> FAP(4B9) VL]]> <![CDATA[<400> 83]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Thr Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Ile Met Leu Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 100 105 <![CDATA[<210> 84]]> <![CDATA[<211> 760]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 84]]> Met Lys Thr Trp Val Lys Ile Val Phe Gly Val Ala Thr Ser Ala Val 1 5 10 15 Leu Ala Leu Leu Val Met Cys Ile Val Leu Arg Pro Ser Arg Val His 20 25 30 Asn Ser Glu Glu Asn Thr Met Arg Ala Leu Thr Leu Lys Asp Ile Leu 35 40 45 Asn Gly Thr Phe Ser Tyr Lys Thr Phe Phe Pro Asn Trp Ile Ser Gly 50 55 60 Gln Glu Tyr Leu His Gln Ser Ala Asp Asn Asn Ile Val Leu Tyr Asn 65 70 75 80 Ile Glu Thr Gly Gln Ser Tyr Thr Ile Leu Ser Asn Arg Thr Met Lys 85 90 95 Ser Val Asn Ala Ser Asn Tyr Gly Leu Ser Pro Asp Arg Gln Phe Val 100 105 110 Tyr Leu Glu Ser Asp Tyr Ser Lys Leu Trp Arg Tyr Ser Tyr Thr Ala 115 120 125 Thr Tyr Tyr Ile Tyr Asp Leu Ser Asn Gly Glu Phe Val Arg Gly Asn 130 135 140 Glu Leu Pro Arg Pro Ile Gln Tyr Leu Cys Trp Ser Pro Val Gly Ser 145 150 155 160 Lys Leu Ala Tyr Val Tyr Gln Asn Asn Ile Tyr Leu Lys Gln Arg Pro 165 170 175 Gly Asp Pro Pro Phe Gln Ile Thr Phe Asn Gly Arg Glu Asn Lys Ile 180 185 190 Phe Asn Gly Ile Pro Asp Trp Val Tyr Glu Glu Glu Met Leu Ala Thr 195 200 205 Lys Tyr Ala Leu Trp Trp Ser Pro Asn Gly Lys Phe Leu Ala Tyr Ala 210 215 220 Glu Phe Asn Asp Thr Asp Ile Pro Val Ile Ala Tyr Ser Tyr Tyr Gly 225 230 235 240 Asp Glu Gln Tyr Pro Arg Thr Ile Asn Ile Pro Tyr Pro Lys Ala Gly 245 250 255 Ala Lys Asn Pro Val Val Arg Ile Phe Ile Ile Asp Thr Thr Tyr Pro 260 265 270 Ala Tyr Val Gly Pro Gln Glu Val Pro Val Pro Ala Met Ile Ala Ser 275 280 285 Ser Asp Tyr Tyr Phe Ser Trp Leu Thr Trp Val Thr Asp Glu Arg Val 290 295 300 Cys Leu Gln Trp Leu Lys Arg Val Gln Asn Val Ser Val Leu Ser Ile 305 310 315 320 Cys Asp Phe Arg Glu Asp Trp Gln Thr Trp Asp Cys Pro Lys Thr Gln 325 330 335 Glu His Ile Glu Glu Ser Arg Thr Gly Trp Ala Gly Gly Phe Phe Val 340 345 350 Ser Thr Pro Val Phe Ser Tyr Asp Ala Ile Ser Tyr Tyr Lys Ile Phe 355 360 365 Ser Asp Lys Asp Gly Tyr Lys His Ile His Tyr Ile Lys Asp Thr Val 370 375 380 Glu Asn Ala Ile Gln Ile Thr Ser Gly Lys Trp Glu Ala Ile Asn Ile 385 390 395 400 Phe Arg Val Thr Gln Asp Ser Leu Phe Tyr Ser Ser Asn Glu Phe Glu 405 410 415 Glu Tyr Pro Gly Arg Arg Asn Ile Tyr Arg Ile Ser Ile Gly Ser Tyr 420 425 430 Pro Pro Ser Lys Lys Cys Val Thr Cys His Leu Arg Lys Glu Arg Cys 435 440 445 Gln Tyr Tyr Thr Ala Ser Phe Ser Asp Tyr Ala Lys Tyr Tyr Ala Leu 450 455 460 Val Cys Tyr Gly Pro Gly Ile Pro Ile Ser Thr Leu His Asp Gly Arg 465 470 475 480 Thr Asp Gln Glu Ile Lys Ile Leu Glu Glu Asn Lys Glu Leu Glu Asn 485 490 495 Ala Leu Lys Asn Ile Gln Leu Pro Lys Glu Glu Ile Lys Lys Leu Glu 500 505 510 Val Asp Glu Ile Thr Leu Trp Tyr Lys Met Ile Leu Pro Pro Gln Phe 515 520 525 Asp Arg Ser Lys Lys Tyr Pro Leu Leu Ile Gln Val Tyr Gly Gly Pro 530 535 540 Cys Ser Gln Ser Val Arg Ser Val Phe Ala Val Asn Trp Ile Ser Tyr 545 550 555 560 Leu Ala Ser Lys Glu Gly Met Val Ile Ala Leu Val Asp Gly Arg Gly 565 570 575 Thr Ala Phe Gln Gly Asp Lys Leu Leu Tyr Ala Val Tyr Arg Lys Leu 580 585 590 Gly Val Tyr Glu Val Glu Asp Gln Ile Thr Ala Val Arg Lys Phe Ile 595 600 605 Glu Met Gly Phe Ile Asp Glu Lys Arg Ile Ala Ile Trp Gly Trp Ser 610 615 620 Tyr Gly Gly Tyr Val Ser Ser Leu Ala Leu Ala Ser Gly Thr Gly Leu 625 630 635 640 Phe Lys Cys Gly Ile Ala Val Ala Pro Val Ser Ser Trp Glu Tyr Tyr 645 650 655 Ala Ser Val Tyr Thr Glu Arg Phe Met Gly Leu Pro Thr Lys Asp Asp 660 665 670 Asn Leu Glu His Tyr Lys Asn Ser Thr Val Met Ala Arg Ala Glu Tyr 675 680 685 Phe Arg Asn Val Asp Tyr Leu Leu Ile His Gly Thr Ala Asp Asp Asn 690 695 700 Val His Phe Gln Asn Ser Ala Gln Ile Ala Lys Ala Leu Val Asn Ala 705 710 715 720 Gln Val Asp Phe Gln Ala Met Trp Tyr Ser Asp Gln Asn His Gly Leu 725 730 735 Ser Gly Leu Ser Thr Asn His Leu Tyr Thr His Met Thr His Phe Leu 740 745 750 Lys Gln Cys Phe Ser Leu Ser Asp 755 760 <![CDATA[<210> 85]]> <![CDATA[<211> 748]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> hu FAP ectodomain+poly-lys-tag+his6-tag]]> <![CDATA[<400> 85]]> Arg Pro Ser Arg Val His Asn Ser Glu Glu Asn Thr Met Arg Ala Leu 1 5 10 15 Thr Leu Lys Asp Ile Leu Asn Gly Thr Phe Ser Tyr Lys Thr Phe Phe 20 25 30 Pro Asn Trp Ile Ser Gly Gln Glu Tyr Leu His Gln Ser Ala Asp Asn 35 40 45 Asn Ile Val Leu Tyr Asn Ile Glu Thr Gly Gln Ser Tyr Thr Ile Leu 50 55 60 Ser Asn Arg Thr Met Lys Ser Val Asn Ala Ser Asn Tyr Gly Leu Ser 65 70 75 80 Pro Asp Arg Gln Phe Val Tyr Leu Glu Ser Asp Tyr Ser Lys Leu Trp 85 90 95 Arg Tyr Ser Tyr Thr Ala Thr Tyr Tyr Ile Tyr Asp Leu Ser Asn Gly 100 105 110 Glu Phe Val Arg Gly Asn Glu Leu Pro Arg Pro Ile Gln Tyr Leu Cys 115 120 125 Trp Ser Pro Val Gly Ser Lys Leu Ala Tyr Val Tyr Gln Asn Asn Ile 130 135 140 Tyr Leu Lys Gln Arg Pro Gly Asp Pro Pro Phe Gln Ile Thr Phe Asn 145 150 155 160 Gly Arg Glu Asn Lys Ile Phe Asn Gly Ile Pro Asp Trp Val Tyr Glu 165 170 175 Glu Glu Met Leu Ala Thr Lys Tyr Ala Leu Trp Trp Ser Pro Asn Gly 180 185 190 Lys Phe Leu Ala Tyr Ala Glu Phe Asn Asp Thr Asp Ile Pro Val Ile 195 200 205 Ala Tyr Ser Tyr Tyr Gly Asp Glu Gln Tyr Pro Arg Thr Ile Asn Ile 210 215 220 Pro Tyr Pro Lys Ala Gly Ala Lys Asn Pro Val Val Arg Ile Phe Ile 225 230 235 240 Ile Asp Thr Thr Tyr Pro Ala Tyr Val Gly Pro Gln Glu Val Pro Val 245 250 255 Pro Ala Met Ile Ala Ser Ser Asp Tyr Tyr Phe Ser Trp Leu Thr Trp 260 265 270 Val Thr Asp Glu Arg Val Cys Leu Gln Trp Leu Lys Arg Val Gln Asn 275 280 285 Val Ser Val Leu Ser Ile Cys Asp Phe Arg Glu Asp Trp Gln Thr Trp 290 295 300 Asp Cys Pro Lys Thr Gln Glu His Ile Glu Glu Ser Arg Thr Gly Trp 305 310 315 320 Ala Gly Gly Phe Phe Val Ser Thr Pro Val Phe Ser Tyr Asp Ala Ile 325 330 335 Ser Tyr Tyr Lys Ile Phe Ser Asp Lys Asp Gly Tyr Lys His Ile His 340 345 350 Tyr Ile Lys Asp Thr Val Glu Asn Ala Ile Gln Ile Thr Ser Gly Lys 355 360 365 Trp Glu Ala Ile Asn Ile Phe Arg Val Thr Gln Asp Ser Leu Phe Tyr 370 375 380 Ser Ser Asn Glu Phe Glu Glu Tyr Pro Gly Arg Arg Asn Ile Tyr Arg 385 390 395 400 Ile Ser Ile Gly Ser Tyr Pro Pro Ser Lys Lys Cys Val Thr Cys His 405 410 415 Leu Arg Lys Glu Arg Cys Gln Tyr Tyr Thr Ala Ser Phe Ser Asp Tyr 420 425 430 Ala Lys Tyr Tyr Ala Leu Val Cys Tyr Gly Pro Gly Ile Pro Ile Ser 435 440 445 Thr Leu His Asp Gly Arg Thr Asp Gln Glu Ile Lys Ile Leu Glu Glu 450 455 460 Asn Lys Glu Leu Glu Asn Ala Leu Lys Asn Ile Gln Leu Pro Lys Glu 465 470 475 480 Glu Ile Lys Lys Leu Glu Val Asp Glu Ile Thr Leu Trp Tyr Lys Met 485 490 495 Ile Leu Pro Pro Gln Phe Asp Arg Ser Lys Lys Tyr Pro Leu Leu Ile 500 505 510 Gln Val Tyr Gly Gly Pro Cys Ser Gln Ser Val Arg Ser Val Phe Ala 515 520 525 Val Asn Trp Ile Ser Tyr Leu Ala Ser Lys Glu Gly Met Val Ile Ala 530 535 540 Leu Val Asp Gly Arg Gly Thr Ala Phe Gln Gly Asp Lys Leu Leu Tyr 545 550 555 560 Ala Val Tyr Arg Lys Leu Gly Val Tyr Glu Val Glu Asp Gln Ile Thr 565 570 575 Ala Val Arg Lys Phe Ile Glu Met Gly Phe Ile Asp Glu Lys Arg Ile 580 585 590 Ala Ile Trp Gly Trp Ser Tyr Gly Gly Tyr Val Ser Ser Leu Ala Leu 595 600 605 Ala Ser Gly Thr Gly Leu Phe Lys Cys Gly Ile Ala Val Ala Pro Val 610 615 620 Ser Ser Trp Glu Tyr Tyr Ala Ser Val Tyr Thr Glu Arg Phe Met Gly 625 630 635 640 Leu Pro Thr Lys Asp Asp Asn Leu Glu His Tyr Lys Asn Ser Thr Val 645 650 655 Met Ala Arg Ala Glu Tyr Phe Arg Asn Val Asp Tyr Leu Leu Ile His 660 665 670 Gly Thr Ala Asp Asp Asn Val His Phe Gln Asn Ser Ala Gln Ile Ala 675 680 685 Lys Ala Leu Val Asn Ala Gln Val Asp Phe Gln Ala Met Trp Tyr Ser 690 695 700 Asp Gln Asn His Gly Leu Ser Gly Leu Ser Thr Asn His Leu Tyr Thr 705 710 715 720 His Met Thr His Phe Leu Lys Gln Cys Phe Ser Leu Ser Asp Gly Lys 725 730 735 Lys Lys Lys Lys Lys Gly His His His His His His 740 745 <![CDATA[<210> 86]]> <![CDATA[<211> 2244]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> hu FAP胞外域+聚-lys-標籤+his6-標籤]]> <![CDATA[<400> 86]]> cgcccttcaa gagttcataa ctctgaagaa aatacaatga gagcactcac actgaaggat 60 attttaaatg gaacattttc ttataaaaca ttttttccaa actggatttc aggacaagaa 120 tatcttcatc aatctgcaga taacaatata gtactttata atattgaaac aggacaatca 180 tataccattt tgagtaatag aaccatgaaa agtgtgaatg cttcaaatta cggcttatca 240 cctgatcggc aatttgtata tctagaaagt gattattcaa agctttggag atactcttac 300 acagcaacat attacatcta tgaccttagc aatggagaat ttgtaagagg aaatgagctt 360 cctcgtccaa ttcagtattt atgctggtcg cctgttggga gtaaattagc atatgtctat 420 caaaacaata tctatttgaa acaaagacca ggagatccac cttttcaaat aacatttaat 480 ggaagagaaa ataaaatatt taatggaatc ccagactggg tttatgaaga ggaaatgctt 540 gctacaaaat atgctctctg gtggtctcct aatggaaaat ttttggcata tgcggaattt 600 aatgatacgg atataccagt tattgcctat tcctattatg gcgatgaaca atatcctaga 660 acaataaata ttccataccc aaaggctgga gctaagaatc ccgttgttcg gatatttatt 720 atcgatacca cttaccctgc gtatgtaggt ccccaggaag tgcctgttcc agcaatgata 780 gcctcaagtg attattattt cagttggctc acgtgggtta ctgatgaacg agtatgtttg 840 cagtggctaa aaagagtcca gaatgtttcg gtcctgtcta tatgtgactt cagggaagac 900 tggcagacat gggattgtcc aaagacccag gagcatatag aagaaagcag aactggatgg 960 gctggtggat tctttgtttc aacaccagtt ttcagctatg atgccatttc gtactacaaa 1020 atatttagtg acaaggatgg ctacaaacat attcactata tcaaagacac tgtggaaaat 1080 gctattcaaa ttacaagtgg caagtgggag gccataaata tattcagagt aacacaggat 1140 tcactgtttt attctagcaa tgaatttgaa gaataccctg gaagaagaaa catctacaga 1200 attagcattg gaagctatcc tccaagcaag aagtgtgtta cttgccatct aaggaaagaa 1260 aggtgccaat attacacagc aagtttcagc gactacgcca agtactatgc acttgtctgc 1320 tacggcccag gcatccccat ttccaccctt catgatggac gcactgatca agaaattaaa 1380 atcctggaag aaaacaagga attggaaaat gctttgaaaa atatccagct gcctaaagag 1440 gaaattaaga aacttgaagt agatgaaatt actttatggt acaagatgat tcttcctcct 1500 caatttgaca gatcaaagaa gtatcccttg ctaattcaag tgtatggtgg tccctgcagt 1560 cagagtgtaa ggtctgtatt tgctgttaat tggatatctt atcttgcaag taaggaaggg 1620 atggtcattg ccttggtgga tggtcgagga acagctttcc aaggtgacaa actcctctat 1680 gcagtgtatc gaaagctggg tgtttatgaa gttgaagacc agattacagc tgtcagaaaa 1740 ttcatagaaa tgggtttcat tgatgaaaaa agaatagcca tatggggctg gtcctatgga 1800 ggatacgttt catcactggc ccttgcatct ggaactggtc ttttcaaatg tggtatagca 1860 gtggctccag tctccagctg ggaatattac gcgtctgtct acacagagag attcatgggt 1920 ctcccaacaa aggatgataa tcttgagcac tataagaatt caactgtgat ggcaagagca 1980 gaatatttca gaaatgtaga ctatcttctc atccacggaa cagcagatga taatgtgcac 2040 tttcaaaact cagcacagat tgctaaagct ctggttaatg cacaagtgga tttccaggca 2100 atgtggtact ctgaccagaa ccacggctta tccggcctgt ccacgaacca cttatacacc 2160 cacatgaccc acttcctaaa gcagtgtttc tctttgtcag acggcaaaaa gaaaaagaaa 2220 aagggccacc accatcacca tcac 2244 <![CDATA[<210> 87]]> <![CDATA[<211> 761]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 小家鼠]]> <![CDATA[<400> 87]]> Met Lys Thr Trp Leu Lys Thr Val Phe Gly Val Thr Thr Leu Ala Ala 1 5 10 15 Leu Ala Leu Val Val Ile Cys Ile Val Leu Arg Pro Ser Arg Val Tyr 20 25 30 Lys Pro Glu Gly Asn Thr Lys Arg Ala Leu Thr Leu Lys Asp Ile Leu 35 40 45 Asn Gly Thr Phe Ser Tyr Lys Thr Tyr Phe Pro Asn Trp Ile Ser Glu 50 55 60 Gln Glu Tyr Leu His Gln Ser Glu Asp Asp Asn Ile Val Phe Tyr Asn 65 70 75 80 Ile Glu Thr Arg Glu Ser Tyr Ile Ile Leu Ser Asn Ser Thr Met Lys 85 90 95 Ser Val Asn Ala Thr Asp Tyr Gly Leu Ser Pro Asp Arg Gln Phe Val 100 105 110 Tyr Leu Glu Ser Asp Tyr Ser Lys Leu Trp Arg Tyr Ser Tyr Thr Ala 115 120 125 Thr Tyr Tyr Ile Tyr Asp Leu Gln Asn Gly Glu Phe Val Arg Gly Tyr 130 135 140 Glu Leu Pro Arg Pro Ile Gln Tyr Leu Cys Trp Ser Pro Val Gly Ser 145 150 155 160 Lys Leu Ala Tyr Val Tyr Gln Asn Asn Ile Tyr Leu Lys Gln Arg Pro 165 170 175 Gly Asp Pro Pro Phe Gln Ile Thr Tyr Thr Gly Arg Glu Asn Arg Ile 180 185 190 Phe Asn Gly Ile Pro Asp Trp Val Tyr Glu Glu Glu Met Leu Ala Thr 195 200 205 Lys Tyr Ala Leu Trp Trp Ser Pro Asp Gly Lys Phe Leu Ala Tyr Val 210 215 220 Glu Phe Asn Asp Ser Asp Ile Pro Ile Ile Ala Tyr Ser Tyr Tyr Gly 225 230 235 240 Asp Gly Gln Tyr Pro Arg Thr Ile Asn Ile Pro Tyr Pro Lys Ala Gly 245 250 255 Ala Lys Asn Pro Val Val Arg Val Phe Ile Val Asp Thr Thr Tyr Pro 260 265 270 His His Val Gly Pro Met Glu Val Pro Val Pro Glu Met Ile Ala Ser 275 280 285 Ser Asp Tyr Tyr Phe Ser Trp Leu Thr Trp Val Ser Ser Glu Arg Val 290 295 300 Cys Leu Gln Trp Leu Lys Arg Val Gln Asn Val Ser Val Leu Ser Ile 305 310 315 320 Cys Asp Phe Arg Glu Asp Trp His Ala Trp Glu Cys Pro Lys Asn Gln 325 330 335 Glu His Val Glu Glu Ser Arg Thr Gly Trp Ala Gly Gly Phe Phe Val 340 345 350 Ser Thr Pro Ala Phe Ser Gln Asp Ala Thr Ser Tyr Tyr Lys Ile Phe 355 360 365 Ser Asp Lys Asp Gly Tyr Lys His Ile His Tyr Ile Lys Asp Thr Val 370 375 380 Glu Asn Ala Ile Gln Ile Thr Ser Gly Lys Trp Glu Ala Ile Tyr Ile 385 390 395 400 Phe Arg Val Thr Gln Asp Ser Leu Phe Tyr Ser Ser Asn Glu Phe Glu 405 410 415 Gly Tyr Pro Gly Arg Arg Asn Ile Tyr Arg Ile Ser Ile Gly Asn Ser 420 425 430 Pro Pro Ser Lys Lys Cys Val Thr Cys His Leu Arg Lys Glu Arg Cys 435 440 445 Gln Tyr Tyr Thr Ala Ser Phe Ser Tyr Lys Ala Lys Tyr Tyr Ala Leu 450 455 460 Val Cys Tyr Gly Pro Gly Leu Pro Ile Ser Thr Leu His Asp Gly Arg 465 470 475 480 Thr Asp Gln Glu Ile Gln Val Leu Glu Glu Asn Lys Glu Leu Glu Asn 485 490 495 Ser Leu Arg Asn Ile Gln Leu Pro Lys Val Glu Ile Lys Lys Leu Lys 500 505 510 Asp Gly Gly Leu Thr Phe Trp Tyr Lys Met Ile Leu Pro Pro Gln Phe 515 520 525 Asp Arg Ser Lys Lys Tyr Pro Leu Leu Ile Gln Val Tyr Gly Gly Pro 530 535 540 Cys Ser Gln Ser Val Lys Ser Val Phe Ala Val Asn Trp Ile Thr Tyr 545 550 555 560 Leu Ala Ser Lys Glu Gly Ile Val Ile Ala Leu Val Asp Gly Arg Gly 565 570 575 Thr Ala Phe Gln Gly Asp Lys Phe Leu His Ala Val Tyr Arg Lys Leu 580 585 590 Gly Val Tyr Glu Val Glu Asp Gln Leu Thr Ala Val Arg Lys Phe Ile 595 600 605 Glu Met Gly Phe Ile Asp Glu Glu Arg Ile Ala Ile Trp Gly Trp Ser 610 615 620 Tyr Gly Gly Tyr Val Ser Ser Leu Ala Leu Ala Ser Gly Thr Gly Leu 625 630 635 640 Phe Lys Cys Gly Ile Ala Val Ala Pro Val Ser Ser Trp Glu Tyr Tyr 645 650 655 Ala Ser Ile Tyr Ser Glu Arg Phe Met Gly Leu Pro Thr Lys Asp Asp 660 665 670 Asn Leu Glu His Tyr Lys Asn Ser Thr Val Met Ala Arg Ala Glu Tyr 675 680 685 Phe Arg Asn Val Asp Tyr Leu Leu Ile His Gly Thr Ala Asp Asp Asn 690 695 700 Val His Phe Gln Asn Ser Ala Gln Ile Ala Lys Ala Leu Val Asn Ala 705 710 715 720 Gln Val Asp Phe Gln Ala Met Trp Tyr Ser Asp Gln Asn His Gly Ile 725 730 735 Ser Ser Gly Arg Ser Gln Asn His Leu Tyr Thr His Met Thr His Phe 740 745 750 Leu Lys Gln Cys Phe Ser Leu Ser Asp 755 760 <![CDATA[<210> 88]]> <![CDATA[<211> 749]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類FAP胞外域+聚-lys-標籤+his6-標籤]]> <![CDATA[<400> 88]]> Arg Pro Ser Arg Val Tyr Lys Pro Glu Gly Asn Thr Lys Arg Ala Leu 1 5 10 15 Thr Leu Lys Asp Ile Leu Asn Gly Thr Phe Ser Tyr Lys Thr Tyr Phe 20 25 30 Pro Asn Trp Ile Ser Glu Gln Glu Tyr Leu His Gln Ser Glu Asp Asp 35 40 45 Asn Ile Val Phe Tyr Asn Ile Glu Thr Arg Glu Ser Tyr Ile Ile Leu 50 55 60 Ser Asn Ser Thr Met Lys Ser Val Asn Ala Thr Asp Tyr Gly Leu Ser 65 70 75 80 Pro Asp Arg Gln Phe Val Tyr Leu Glu Ser Asp Tyr Ser Lys Leu Trp 85 90 95 Arg Tyr Ser Tyr Thr Ala Thr Tyr Tyr Ile Tyr Asp Leu Gln Asn Gly 100 105 110 Glu Phe Val Arg Gly Tyr Glu Leu Pro Arg Pro Ile Gln Tyr Leu Cys 115 120 125 Trp Ser Pro Val Gly Ser Lys Leu Ala Tyr Val Tyr Gln Asn Asn Ile 130 135 140 Tyr Leu Lys Gln Arg Pro Gly Asp Pro Pro Phe Gln Ile Thr Tyr Thr 145 150 155 160 Gly Arg Glu Asn Arg Ile Phe Asn Gly Ile Pro Asp Trp Val Tyr Glu 165 170 175 Glu Glu Met Leu Ala Thr Lys Tyr Ala Leu Trp Trp Ser Pro Asp Gly 180 185 190 Lys Phe Leu Ala Tyr Val Glu Phe Asn Asp Ser Asp Ile Pro Ile Ile 195 200 205 Ala Tyr Ser Tyr Tyr Gly Asp Gly Gln Tyr Pro Arg Thr Ile Asn Ile 210 215 220 Pro Tyr Pro Lys Ala Gly Ala Lys Asn Pro Val Val Arg Val Phe Ile 225 230 235 240 Val Asp Thr Thr Tyr Pro His His Val Gly Pro Met Glu Val Pro Val 245 250 255 Pro Glu Met Ile Ala Ser Ser Asp Tyr Tyr Phe Ser Trp Leu Thr Trp 260 265 270 Val Ser Ser Glu Arg Val Cys Leu Gln Trp Leu Lys Arg Val Gln Asn 275 280 285 Val Ser Val Leu Ser Ile Cys Asp Phe Arg Glu Asp Trp His Ala Trp 290 295 300 Glu Cys Pro Lys Asn Gln Glu His Val Glu Glu Ser Arg Thr Gly Trp 305 310 315 320 Ala Gly Gly Phe Phe Val Ser Thr Pro Ala Phe Ser Gln Asp Ala Thr 325 330 335 Ser Tyr Tyr Lys Ile Phe Ser Asp Lys Asp Gly Tyr Lys His Ile His 340 345 350 Tyr Ile Lys Asp Thr Val Glu Asn Ala Ile Gln Ile Thr Ser Gly Lys 355 360 365 Trp Glu Ala Ile Tyr Ile Phe Arg Val Thr Gln Asp Ser Leu Phe Tyr 370 375 380 Ser Ser Asn Glu Phe Glu Gly Tyr Pro Gly Arg Arg Asn Ile Tyr Arg 385 390 395 400 Ile Ser Ile Gly Asn Ser Pro Pro Ser Lys Lys Cys Val Thr Cys His 405 410 415 Leu Arg Lys Glu Arg Cys Gln Tyr Tyr Thr Ala Ser Phe Ser Tyr Lys 420 425 430 Ala Lys Tyr Tyr Ala Leu Val Cys Tyr Gly Pro Gly Leu Pro Ile Ser 435 440 445 Thr Leu His Asp Gly Arg Thr Asp Gln Glu Ile Gln Val Leu Glu Glu 450 455 460 Asn Lys Glu Leu Glu Asn Ser Leu Arg Asn Ile Gln Leu Pro Lys Val 465 470 475 480 Glu Ile Lys Lys Leu Lys Asp Gly Gly Leu Thr Phe Trp Tyr Lys Met 485 490 495 Ile Leu Pro Pro Gln Phe Asp Arg Ser Lys Lys Tyr Pro Leu Leu Ile 500 505 510 Gln Val Tyr Gly Gly Pro Cys Ser Gln Ser Val Lys Ser Val Phe Ala 515 520 525 Val Asn Trp Ile Thr Tyr Leu Ala Ser Lys Glu Gly Ile Val Ile Ala 530 535 540 Leu Val Asp Gly Arg Gly Thr Ala Phe Gln Gly Asp Lys Phe Leu His 545 550 555 560 Ala Val Tyr Arg Lys Leu Gly Val Tyr Glu Val Glu Asp Gln Leu Thr 565 570 575 Ala Val Arg Lys Phe Ile Glu Met Gly Phe Ile Asp Glu Glu Arg Ile 580 585 590 Ala Ile Trp Gly Trp Ser Tyr Gly Gly Tyr Val Ser Ser Leu Ala Leu 595 600 605 Ala Ser Gly Thr Gly Leu Phe Lys Cys Gly Ile Ala Val Ala Pro Val 610 615 620 Ser Ser Trp Glu Tyr Tyr Ala Ser Ile Tyr Ser Glu Arg Phe Met Gly 625 630 635 640 Leu Pro Thr Lys Asp Asp Asn Leu Glu His Tyr Lys Asn Ser Thr Val 645 650 655 Met Ala Arg Ala Glu Tyr Phe Arg Asn Val Asp Tyr Leu Leu Ile His 660 665 670 Gly Thr Ala Asp Asp Asn Val His Phe Gln Asn Ser Ala Gln Ile Ala 675 680 685 Lys Ala Leu Val Asn Ala Gln Val Asp Phe Gln Ala Met Trp Tyr Ser 690 695 700 Asp Gln Asn His Gly Ile Leu Ser Gly Arg Ser Gln Asn His Leu Tyr 705 710 715 720 Thr His Met Thr His Phe Leu Lys Gln Cys Phe Ser Leu Ser Asp Gly 725 730 735 Lys Lys Lys Lys Lys Lys Gly His His His His His His 740 745 <![CDATA[<210> 89]]> <![CDATA[<211> 2247]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類FAP胞外域+聚-lys-標籤+his6-標籤]]> <![CDATA[<400> 89]]> cgtccctcaa gagtttacaa acctgaagga aacacaaaga gagctcttac cttgaaggat 60 attttaaatg gaacattctc atataaaaca tattttccca actggatttc agaacaagaa 120 tatcttcatc aatctgagga tgataacata gtattttata atattgaaac aagagaatca 180 tatatcattt tgagtaatag caccatgaaa agtgtgaatg ctacagatta tggtttgtca 240 cctgatcggc aatttgtgta tctagaaagt gattattcaa agctctggcg atattcatac 300 acagcgacat actacatcta cgaccttcag aatggggaat ttgtaagagg atacgagctc 360 cctcgtccaa ttcagtatct atgctggtcg cctgttggga gtaaattagc atatgtatat 420 caaaacaata tttatttgaa acaaagacca ggagatccac cttttcaaat aacttatact 480 ggaagagaaa atagaatatt taatggaata ccagactggg tttatgaaga ggaaatgctt 540 gccacaaaat atgctctttg gtggtctcca gatggaaaat ttttggcata tgtagaattt 600 aatgattcag atataccaat tattgcctat tcttattatg gtgatggaca gtatcctaga 660 actataaata ttccatatcc aaaggctggg gctaagaatc cggttgttcg tgtttttatt 720 gttgacacca cctaccctca ccacgtgggc ccaatggaag tgccagttcc agaaatgata 780 gcctcaagtg actattattt cagctggctc acatgggtgt ccagtgaacg agtatgcttg 840 cagtggctaa aaagagtgca gaatgtctca gtcctgtcta tatgtgattt cagggaagac 900 tggcatgcat gggaatgtcc aaagaaccag gagcatgtag aagaaagcag aacaggatgg 960 gctggtggat tctttgtttc gacaccagct tttagccagg atgccacttc ttactacaaa 1020 atatttagcg acaaggatgg ttacaaacat attcactaca tcaaagacac tgtggaaaat 1080 gctattcaaa ttacaagtgg caagtgggag gccatatata tattccgcgt aacacaggat 1140 tcactgtttt attctagcaa tgaatttgaa ggttaccctg gaagaagaaa catctacaga 1200 attagcattg gaaactctcc tccgagcaag aagtgtgtta cttgccatct aaggaaagaa 1260 aggtgccaat attacacagc aagtttcagc tacaaagcca agtactatgc actcgtctgc 1320 tatggccctg gcctccccat ttccaccctc catgatggcc gcacagacca agaaatacaa 1380 gtattagaag aaaacaaaga actggaaaat tctctgagaa atatccagct gcctaaagtg 1440 gagattaaga agctcaaaga cgggggactg actttctggt acaagatgat tctgcctcct 1500 cagtttgaca gatcaaagaa gtaccctttg ctaattcaag tgtatggtgg tccttgtagc 1560 cagagtgtta agtctgtgtt tgctgttaat tggataactt atctcgcaag taaggagggg 1620 atagtcattg ccctggtaga tggtcggggc actgctttcc aaggtgacaa attcctgcat 1680 gccgtgtatc gaaaactggg tgtatatgaa gttgaggacc agctcacagc tgtcagaaaa 1740 ttcatagaaa tgggtttcat tgatgaagaa agaatagcca tatggggctg gtcctacgga 1800 ggttatgttt catccctggc ccttgcatct ggaactggtc ttttcaaatg tggcatagca 1860 gtggctccag tctccagctg ggaatattac gcatctatct actcagagag attcatgggc 1920 ctcccaacaa aggacgacaa tctcgaacac tataaaaatt caactgtgat ggcaagagca 1980 gaatatttca gaaatgtaga ctatcttctc atccacggaa cagcagatga taatgtgcac 2040 tttcagaact cagcacagat tgctaaagct ttggttaatg cacaagtgga tttccaggcg 2100 atgtggtact ctgaccagaa ccatggtata ttatctgggc gctcccagaa tcatttatat 2160 acccacatga cgcacttcct caagcaatgc ttttctttat cagacggcaa aaagaaaaag 2220 aaaaagggcc accaccatca ccatcac 2247 <![CDATA[<210> 90]]> <![CDATA[<211> 748]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 獼猴FAP胞外域+聚-lys-標籤+his6-標籤]]> <![CDATA[<400> 90]]> Arg Pro Pro Arg Val His Asn Ser Glu Glu Asn Thr Met Arg Ala Leu 1 5 10 15 Thr Leu Lys Asp Ile Leu Asn Gly Thr Phe Ser Tyr Lys Thr Phe Phe 20 25 30 Pro Asn Trp Ile Ser Gly Gln Glu Tyr Leu His Gln Ser Ala Asp Asn 35 40 45 Asn Ile Val Leu Tyr Asn Ile Glu Thr Gly Gln Ser Tyr Thr Ile Leu 50 55 60 Ser Asn Arg Thr Met Lys Ser Val Asn Ala Ser Asn Tyr Gly Leu Ser 65 70 75 80 Pro Asp Arg Gln Phe Val Tyr Leu Glu Ser Asp Tyr Ser Lys Leu Trp 85 90 95 Arg Tyr Ser Tyr Thr Ala Thr Tyr Tyr Ile Tyr Asp Leu Ser Asn Gly 100 105 110 Glu Phe Val Arg Gly Asn Glu Leu Pro Arg Pro Ile Gln Tyr Leu Cys 115 120 125 Trp Ser Pro Val Gly Ser Lys Leu Ala Tyr Val Tyr Gln Asn Asn Ile 130 135 140 Tyr Leu Lys Gln Arg Pro Gly Asp Pro Pro Phe Gln Ile Thr Phe Asn 145 150 155 160 Gly Arg Glu Asn Lys Ile Phe Asn Gly Ile Pro Asp Trp Val Tyr Glu 165 170 175 Glu Glu Met Leu Ala Thr Lys Tyr Ala Leu Trp Trp Ser Pro Asn Gly 180 185 190 Lys Phe Leu Ala Tyr Ala Glu Phe Asn Asp Thr Asp Ile Pro Val Ile 195 200 205 Ala Tyr Ser Tyr Tyr Gly Asp Glu Gln Tyr Pro Arg Thr Ile Asn Ile 210 215 220 Pro Tyr Pro Lys Ala Gly Ala Lys Asn Pro Phe Val Arg Ile Phe Ile 225 230 235 240 Ile Asp Thr Thr Tyr Pro Ala Tyr Val Gly Pro Gln Glu Val Pro Val 245 250 255 Pro Ala Met Ile Ala Ser Ser Asp Tyr Tyr Phe Ser Trp Leu Thr Trp 260 265 270 Val Thr Asp Glu Arg Val Cys Leu Gln Trp Leu Lys Arg Val Gln Asn 275 280 285 Val Ser Val Leu Ser Ile Cys Asp Phe Arg Glu Asp Trp Gln Thr Trp 290 295 300 Asp Cys Pro Lys Thr Gln Glu His Ile Glu Glu Ser Arg Thr Gly Trp 305 310 315 320 Ala Gly Gly Phe Phe Val Ser Thr Pro Val Phe Ser Tyr Asp Ala Ile 325 330 335 Ser Tyr Tyr Lys Ile Phe Ser Asp Lys Asp Gly Tyr Lys His Ile His 340 345 350 Tyr Ile Lys Asp Thr Val Glu Asn Ala Ile Gln Ile Thr Ser Gly Lys 355 360 365 Trp Glu Ala Ile Asn Ile Phe Arg Val Thr Gln Asp Ser Leu Phe Tyr 370 375 380 Ser Ser Asn Glu Phe Glu Asp Tyr Pro Gly Arg Arg Asn Ile Tyr Arg 385 390 395 400 Ile Ser Ile Gly Ser Tyr Pro Pro Ser Lys Lys Cys Val Thr Cys His 405 410 415 Leu Arg Lys Glu Arg Cys Gln Tyr Tyr Thr Ala Ser Phe Ser Asp Tyr 420 425 430 Ala Lys Tyr Tyr Ala Leu Val Cys Tyr Gly Pro Gly Ile Pro Ile Ser 435 440 445 Thr Leu His Asp Gly Arg Thr Asp Gln Glu Ile Lys Ile Leu Glu Glu 450 455 460 Asn Lys Glu Leu Glu Asn Ala Leu Lys Asn Ile Gln Leu Pro Lys Glu 465 470 475 480 Glu Ile Lys Lys Leu Glu Val Asp Glu Ile Thr Leu Trp Tyr Lys Met 485 490 495 Ile Leu Pro Pro Gln Phe Asp Arg Ser Lys Lys Tyr Pro Leu Leu Ile 500 505 510 Gln Val Tyr Gly Gly Pro Cys Ser Gln Ser Val Arg Ser Val Phe Ala 515 520 525 Val Asn Trp Ile Ser Tyr Leu Ala Ser Lys Glu Gly Met Val Ile Ala 530 535 540 Leu Val Asp Gly Arg Gly Thr Ala Phe Gln Gly Asp Lys Leu Leu Tyr 545 550 555 560 Ala Val Tyr Arg Lys Leu Gly Val Tyr Glu Val Glu Asp Gln Ile Thr 565 570 575 Ala Val Arg Lys Phe Ile Glu Met Gly Phe Ile Asp Glu Lys Arg Ile 580 585 590 Ala Ile Trp Gly Trp Ser Tyr Gly Gly Tyr Val Ser Ser Leu Ala Leu 595 600 605 Ala Ser Gly Thr Gly Leu Phe Lys Cys Gly Ile Ala Val Ala Pro Val 610 615 620 Ser Ser Trp Glu Tyr Tyr Ala Ser Val Tyr Thr Glu Arg Phe Met Gly 625 630 635 640 Leu Pro Thr Lys Asp Asp Asn Leu Glu His Tyr Lys Asn Ser Thr Val 645 650 655 Met Ala Arg Ala Glu Tyr Phe Arg Asn Val Asp Tyr Leu Leu Ile His 660 665 670 Gly Thr Ala Asp Asp Asn Val His Phe Gln Asn Ser Ala Gln Ile Ala 675 680 685 Lys Ala Leu Val Asn Ala Gln Val Asp Phe Gln Ala Met Trp Tyr Ser 690 695 700 Asp Gln Asn His Gly Leu Ser Gly Leu Ser Thr Asn His Leu Tyr Thr 705 710 715 720 His Met Thr His Phe Leu Lys Gln Cys Phe Ser Leu Ser Asp Gly Lys 725 730 735 Lys Lys Lys Lys Lys Gly His His His His His His 740 745 <![CDATA[<210> 91]]> <![CDATA[<211> 2244]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 獼猴FAP胞外域+聚-lys-標籤+his6-標籤]]> <![CDATA[<400> 91]]> cgccctccaa gagttcataa ctctgaagaa aatacaatga gagcactcac actgaaggat 60 attttaaatg ggacattttc ttataaaaca ttttttccaa actggatttc aggacaagaa 120 tatcttcatc aatctgcaga taacaatata gtactttata atattgaaac aggacaatca 180 tataccattt tgagtaacag aaccatgaaa agtgtgaatg cttcaaatta tggcttatca 240 cctgatcggc aatttgtata tctagaaagt gattattcaa agctttggag atactcttac 300 acagcaacat attacatcta tgaccttagc aatggagaat ttgtaagagg aaatgagctt 360 cctcgtccaa ttcagtattt atgctggtcg cctgttggga gtaaattagc atatgtctat 420 caaaacaata tctatttgaa acaaagacca ggagatccac cttttcaaat aacatttaat 480 ggaagagaaa ataaaatatt taatggaatc ccagactggg tttatgaaga ggaaatgctt 540 gctacaaaat atgctctctg gtggtctcct aatggaaaat ttttggcata tgcggaattt 600 aatgatacag atataccagt tattgcctat tcctattatg gcgatgaaca atatcccaga 660 acaataaata ttccataccc aaaggccgga gctaagaatc cttttgttcg gatatttatt 720 atcgatacca cttaccctgc gtatgtaggt ccccaggaag tgcctgttcc agcaatgata 780 gcctcaagtg attattattt cagttggctc acgtgggtta ctgatgaacg agtatgtttg 840 cagtggctaa aaagagtcca gaatgtttcg gtcttgtcta tatgtgattt cagggaagac 900 tggcagacat gggattgtcc aaagacccag gagcatatag aagaaagcag aactggatgg 960 gctggtggat tctttgtttc aacaccagtt ttcagctatg atgccatttc atactacaaa 1020 atatttagtg acaaggatgg ctacaaacat attcactata tcaaagacac tgtggaaaat 1080 gctattcaaa ttacaagtgg caagtgggag gccataaata tattcagagt aacacaggat 1140 tcactgtttt attctagcaa tgaatttgaa gattaccctg gaagaagaaa catctacaga 1200 attagcattg gaagctatcc tccaagcaag aagtgtgtta cttgccatct aaggaaagaa 1260 aggtgccaat attacacagc aagtttcagc gactacgcca agtactatgc acttgtctgc 1320 tatggcccag gcatccccat ttccaccctt catgacggac gcactgatca agaaattaaa 1380 atcctggaag aaaacaagga attggaaaat gctttgaaaa atatccagct gcctaaagag 1440 gaaattaaga aacttgaagt agatgaaatt actttatggt acaagatgat tcttcctcct 1500 caatttgaca gatcaaagaa gtatcccttg ctaattcaag tgtatggtgg tccctgcagt 1560 cagagtgtaa ggtctgtatt tgctgttaat tggatatctt atcttgcaag taaggaaggg 1620 atggtcattg ccttggtgga tggtcgggga acagctttcc aaggtgacaa actcctgtat 1680 gcagtgtatc gaaagctggg tgtttatgaa gttgaagacc agattacagc tgtcagaaaa 1740 ttcatagaaa tgggtttcat tgatgaaaaa agaatagcca tatggggctg gtcctatgga 1800 ggatatgttt catcactggc ccttgcatct ggaactggtc ttttcaaatg tgggatagca 1860 gtggctccag tctccagctg ggaatattac gcgtctgtct acacagagag attcatgggt 1920 ctcccaacaa aggatgataa tcttgagcac tataagaatt caactgtgat ggcaagagca 1980 gaatatttca gaaatgtaga ctatcttctc atccacggaa cagcagatga taatgtgcac 2040 tttcaaaact cagcacagat tgctaaagct ctggttaatg cacaagtgga tttccaggca 2100 atgtggtact ctgaccagaa ccacggctta tccggcctgt ccacgaacca cttatacacc 2160 cacatgaccc acttcctaaa gcagtgtttc tctttgtcag acggcaaaaa gaaaaagaaa 2220 aagggccacc accatcacca tcac 2244 <![CDATA[<210> 92]]> <![CDATA[<211> 702]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 92]]> Met Glu Ser Pro Ser Ala Pro Pro His Arg Trp Cys Ile Pro Trp Gln 1 5 10 15 Arg Leu Leu Leu Thr Ala Ser Leu Leu Thr Phe Trp Asn Pro Pro Thr 20 25 30 Thr Ala Lys Leu Thr Ile Glu Ser Thr Pro Phe Asn Val Ala Glu Gly 35 40 45 Lys Glu Val Leu Leu Leu Val His Asn Leu Pro Gln His Leu Phe Gly 50 55 60 Tyr Ser Trp Tyr Lys Gly Glu Arg Val Asp Gly Asn Arg Gln Ile Ile 65 70 75 80 Gly Tyr Val Ile Gly Thr Gln Gln Ala Thr Pro Gly Pro Ala Tyr Ser 85 90 95 Gly Arg Glu Ile Ile Tyr Pro Asn Ala Ser Leu Leu Ile Gln Asn Ile 100 105 110 Ile Gln Asn Asp Thr Gly Phe Tyr Thr Leu His Val Ile Lys Ser Asp 115 120 125 Leu Val Asn Glu Glu Ala Thr Gly Gln Phe Arg Val Tyr Pro Glu Leu 130 135 140 Pro Lys Pro Ser Ile Ser Ser Asn Asn Ser Lys Pro Val Glu Asp Lys 145 150 155 160 Asp Ala Val Ala Phe Thr Cys Glu Pro Glu Thr Gln Asp Ala Thr Tyr 165 170 175 Leu Trp Trp Val Asn Asn Gln Ser Leu Pro Val Ser Pro Arg Leu Gln 180 185 190 Leu Ser Asn Gly Asn Arg Thr Leu Thr Leu Phe Asn Val Thr Arg Asn 195 200 205 Asp Thr Ala Ser Tyr Lys Cys Glu Thr Gln Asn Pro Val Ser Ala Arg 210 215 220 Arg Ser Asp Ser Val Ile Leu Asn Val Leu Tyr Gly Pro Asp Ala Pro 225 230 235 240 Thr Ile Ser Pro Leu Asn Thr Ser Tyr Arg Ser Gly Glu Asn Leu Asn 245 250 255 Leu Ser Cys His Ala Ala Ser Asn Pro Pro Ala Gln Tyr Ser Trp Phe 260 265 270 Val Asn Gly Thr Phe Gln Gln Ser Thr Gln Glu Leu Phe Ile Pro Asn 275 280 285 Ile Thr Val Asn Asn Ser Gly Ser Tyr Thr Cys Gln Ala His Asn Ser 290 295 300 Asp Thr Gly Leu Asn Arg Thr Thr Val Thr Thr Ile Thr Val Tyr Ala 305 310 315 320 Glu Pro Pro Lys Pro Phe Ile Thr Ser Asn Asn Ser Asn Pro Val Glu 325 330 335 Asp Glu Asp Ala Val Ala Leu Thr Cys Glu Pro Glu Ile Gln Asn Thr 340 345 350 Thr Tyr Leu Trp Trp Val Asn Asn Gln Ser Leu Pro Val Ser Pro Arg 355 360 365 Leu Gln Leu Ser Asn Asp Asn Arg Thr Leu Thr Leu Leu Ser Val Thr 370 375 380 Arg Asn Asp Val Gly Pro Tyr Glu Cys Gly Ile Gln Asn Lys Leu Ser 385 390 395 400 Val Asp His Ser Asp Pro Val Ile Leu Asn Val Leu Tyr Gly Pro Asp 405 410 415 Asp Pro Thr Ile Ser Pro Ser Tyr Thr Tyr Tyr Arg Pro Gly Val Asn 420 425 430 Leu Ser Leu Ser Cys His Ala Ala Ser Asn Pro Pro Ala Gln Tyr Ser 435 440 445 Trp Leu Ile Asp Gly Asn Ile Gln Gln His Thr Gln Glu Leu Phe Ile 450 455 460 Ser Asn Ile Thr Glu Lys Asn Ser Gly Leu Tyr Thr Cys Gln Ala Asn 465 470 475 480 Asn Ser Ala Ser Gly His Ser Arg Thr Thr Val Lys Thr Ile Thr Val 485 490 495 Ser Ala Glu Leu Pro Lys Pro Ser Ile Ser Ser Asn Asn Ser Lys Pro 500 505 510 Val Glu Asp Lys Asp Ala Val Ala Phe Thr Cys Glu Pro Glu Ala Gln 515 520 525 Asn Thr Thr Tyr Leu Trp Trp Val Asn Gly Gln Ser Leu Pro Val Ser 530 535 540 Pro Arg Leu Gln Leu Ser Asn Gly Asn Arg Thr Leu Thr Leu Phe Asn 545 550 555 560 Val Thr Arg Asn Asp Ala Arg Ala Tyr Val Cys Gly Ile Gln Asn Ser 565 570 575 Val Ser Ala Asn Arg Ser Asp Pro Val Thr Leu Asp Val Leu Tyr Gly 580 585 590 Pro Asp Thr Pro Ile Ile Ser Pro Pro Asp Ser Ser Tyr Leu Ser Gly 595 600 605 Ala Asn Leu Asn Leu Ser Cys His Ser Ala Ser Asn Pro Ser Pro Gln 610 615 620 Tyr Ser Trp Arg Ile Asn Gly Ile Pro Gln Gln His Thr Gln Val Leu 625 630 635 640 Phe Ile Ala Lys Ile Thr Pro Asn Asn Asn Gly Thr Tyr Ala Cys Phe 645 650 655 Val Ser Asn Leu Ala Thr Gly Arg Asn Asn Ser Ile Val Lys Ser Ile 660 665 670 Thr Val Ser Ala Ser Gly Thr Ser Pro Gly Leu Ser Ala Gly Ala Thr 675 680 685 Val Gly Ile Met Ile Gly Val Leu Val Gly Val Ala Leu Ile 690 695 700 <![CDATA[<210> 93]]> <![CDATA[<211> 2322]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 93]]> Met Gln Ser Gly Pro Arg Pro Pro Leu Pro Ala Pro Gly Leu Ala Leu 1 5 10 15 Ala Leu Thr Leu Thr Met Leu Ala Arg Leu Ala Ser Ala Ala Ser Phe 20 25 30 Phe Gly Glu Asn His Leu Glu Val Pro Val Ala Thr Ala Leu Thr Asp 35 40 45 Ile Asp Leu Gln Leu Gln Phe Ser Thr Ser Gln Pro Glu Ala Leu Leu 50 55 60 Leu Leu Ala Ala Gly Pro Ala Asp His Leu Leu Leu Gln Leu Tyr Ser 65 70 75 80 Gly Arg Leu Gln Val Arg Leu Val Leu Gly Gln Glu Glu Leu Arg Leu 85 90 95 Gln Thr Pro Ala Glu Thr Leu Leu Ser Asp Ser Ile Pro His Thr Val 100 105 110 Val Leu Thr Val Val Glu Gly Trp Ala Thr Leu Ser Val Asp Gly Phe 115 120 125 Leu Asn Ala Ser Ser Ala Val Pro Gly Ala Pro Leu Glu Val Pro Tyr 130 135 140 Gly Leu Phe Val Gly Gly Thr Gly Thr Leu Gly Leu Pro Tyr Leu Arg 145 150 155 160 Gly Thr Ser Arg Pro Leu Arg Gly Cys Leu His Ala Ala Thr Leu Asn 165 170 175 Gly Arg Ser Leu Leu Arg Pro Leu Thr Pro Asp Val His Glu Gly Cys 180 185 190 Ala Glu Glu Phe Ser Ala Ser Asp Asp Val Ala Leu Gly Phe Ser Gly 195 200 205 Pro His Ser Leu Ala Ala Phe Pro Ala Trp Gly Thr Gln Asp Glu Gly 210 215 220 Thr Leu Glu Phe Thr Leu Thr Thr Gln Ser Arg Gln Ala Pro Leu Ala 225 230 235 240 Phe Gln Ala Gly Gly Arg Arg Gly Asp Phe Ile Tyr Val Asp Ile Phe 245 250 255 Glu Gly His Leu Arg Ala Val Val Glu Lys Gly Gln Gly Thr Val Leu 260 265 270 Leu His Asn Ser Val Pro Val Ala Asp Gly Gln Pro His Glu Val Ser 275 280 285 Val His Ile Asn Ala His Arg Leu Glu Ile Ser Val Asp Gln Tyr Pro 290 295 300 Thr His Thr Ser Asn Arg Gly Val Leu Ser Tyr Leu Glu Pro Arg Gly 305 310 315 320 Ser Leu Leu Leu Gly Gly Leu Asp Ala Glu Ala Ser Arg His Leu Gln 325 330 335 Glu His Arg Leu Gly Leu Thr Pro Glu Ala Thr Asn Ala Ser Leu Leu 340 345 350 Gly Cys Met Glu Asp Leu Ser Val Asn Gly Gln Arg Arg Gly Leu Arg 355 360 365 Glu Ala Leu Leu Thr Arg Asn Met Ala Ala Gly Cys Arg Leu Glu Glu 370 375 380 Glu Glu Tyr Glu Asp Asp Ala Tyr Gly His Tyr Glu Ala Phe Ser Thr 385 390 395 400 Leu Ala Pro Glu Ala Trp Pro Ala Met Glu Leu Pro Glu Pro Cys Val 405 410 415 Pro Glu Pro Gly Leu Pro Pro Val Phe Ala Asn Phe Thr Gln Leu Leu 420 425 430 Thr Ile Ser Pro Leu Val Val Ala Glu Gly Gly Thr Ala Trp Leu Glu 435 440 445 Trp Arg His Val Gln Pro Thr Leu Asp Leu Met Glu Ala Glu Leu Arg 450 455 460 Lys Ser Gln Val Leu Phe Ser Val Thr Arg Gly Ala Arg His Gly Glu 465 470 475 480 Leu Glu Leu Asp Ile Pro Gly Ala Gln Ala Arg Lys Met Phe Thr Leu 485 490 495 Leu Asp Val Val Asn Arg Lys Ala Arg Phe Ile His Asp Gly Ser Glu 500 505 510 Asp Thr Ser Asp Gln Leu Val Leu Glu Val Ser Val Thr Ala Arg Val 515 520 525 Pro Met Pro Ser Cys Leu Arg Arg Gly Gln Thr Tyr Leu Leu Pro Ile 530 535 540 Gln Val Asn Pro Val Asn Asp Pro Pro His Ile Ile Phe Pro His Gly 545 550 555 560 Ser Leu Met Val Ile Leu Glu His Thr Gln Lys Pro Leu Gly Pro Glu 565 570 575 Val Phe Gln Ala Tyr Asp Pro Asp Ser Ala Cys Glu Gly Leu Thr Phe 580 585 590 Gln Val Leu Gly Thr Ser Ser Gly Leu Pro Val Glu Arg Arg Asp Gln 595 600 605 Pro Gly Glu Pro Ala Thr Glu Phe Ser Cys Arg Glu Leu Glu Ala Gly 610 615 620 Ser Leu Val Tyr Val His Arg Gly Gly Pro Ala Gln Asp Leu Thr Phe 625 630 635 640 Arg Val Ser Asp Gly Leu Gln Ala Ser Pro Pro Ala Thr Leu Lys Val 645 650 655 Val Ala Ile Arg Pro Ala Ile Gln Ile His Arg Ser Thr Gly Leu Arg 660 665 670 Leu Ala Gln Gly Ser Ala Met Pro Ile Leu Pro Ala Asn Leu Ser Val 675 680 685 Glu Thr Asn Ala Val Gly Gln Asp Val Ser Val Leu Phe Arg Val Thr 690 695 700 Gly Ala Leu Gln Phe Gly Glu Leu Gln Lys Gln Gly Ala Gly Gly Val 705 710 715 720 Glu Gly Ala Glu Trp Trp Ala Thr Gln Ala Phe His Gln Arg Asp Val 725 730 735 Glu Gln Gly Arg Val Arg Tyr Leu Ser Thr Asp Pro Gln His His Ala 740 745 750 Tyr Asp Thr Val Glu Asn Leu Ala Leu Glu Val Gln Val Gly Gln Glu 755 760 765 Ile Leu Ser Asn Leu Ser Phe Pro Val Thr Ile Gln Arg Ala Thr Val 770 775 780 Trp Met Leu Arg Leu Glu Pro Leu His Thr Gln Asn Thr Gln Gln Glu 785 790 795 800 Thr Leu Thr Thr Ala His Leu Glu Ala Thr Leu Glu Glu Ala Gly Pro 805 810 815 Ser Pro Pro Thr Phe His Tyr Glu Val Val Gln Ala Pro Arg Lys Gly 820 825 830 Asn Leu Gln Leu Gln Gly Thr Arg Leu Ser Asp Gly Gln Gly Phe Thr 835 840 845 Gln Asp Asp Ile Gln Ala Gly Arg Val Thr Tyr Gly Ala Thr Ala Arg 850 855 860 Ala Ser Glu Ala Val Glu Asp Thr Phe Arg Phe Arg Val Thr Ala Pro 865 870 875 880 Pro Tyr Phe Ser Pro Leu Tyr Thr Phe Pro Ile His Ile Gly Gly Asp 885 890 895 Pro Asp Ala Pro Val Leu Thr Asn Val Leu Leu Val Val Pro Glu Gly 900 905 910 Gly Glu Gly Val Leu Ser Ala Asp His Leu Phe Val Lys Ser Leu Asn 915 920 925 Ser Ala Ser Tyr Leu Tyr Glu Val Met Glu Arg Pro Arg His Gly Arg 930 935 940 Leu Ala Trp Arg Gly Thr Gln Asp Lys Thr Thr Met Val Thr Ser Phe 945 950 955 960 Thr Asn Glu Asp Leu Leu Arg Gly Arg Leu Val Tyr Gln His Asp Asp 965 970 975 Ser Glu Thr Thr Glu Asp Asp Ile Pro Phe Val Ala Thr Arg Gln Gly 980 985 990 Glu Ser Ser Gly Asp Met Ala Trp Glu Glu Val Arg Gly Val Phe Arg 995 1000 1005 Val Ala Ile Gln Pro Val Asn Asp His Ala Pro Val Gln Thr Ile 1010 1015 1020 Ser Arg Ile Phe His Val Ala Arg Gly Gly Arg Arg Leu Leu Thr 1025 1030 1035 Thr Asp Asp Val Ala Phe Ser Asp Ala Asp Ser Gly Phe Ala Asp 1040 1045 1050 Ala Gln Leu Val Leu Thr Arg Lys Asp Leu Leu Phe Gly Ser Ile 1055 1060 1065 Val Ala Val Asp Glu Pro Thr Arg Pro Ile Tyr Arg Phe Thr Gln 1070 1075 1080 Glu Asp Leu Arg Lys Arg Arg Val Leu Phe Val His Ser Gly Ala 1085 1090 1095 Asp Arg Gly Trp Ile Gln Leu Gln Val Ser Asp Gly Gln His Gln 1100 1105 1110 Ala Thr Ala Leu Leu Glu Val Gln Ala Ser Glu Pro Tyr Leu Arg 1115 1120 1125 Val Ala Asn Gly Ser Ser Leu Val Val Pro Gln Gly Gly Gln Gly 1130 1135 1140 Thr Ile Asp Thr Ala Val Leu His Leu Asp Thr Asn Leu Asp Ile 1145 1150 1155 Arg Ser Gly Asp Glu Val His Tyr His Val Thr Ala Gly Pro Arg 1160 1165 1170 Trp Gly Gln Leu Val Arg Ala Gly Gln Pro Ala Thr Ala Phe Ser 1175 1180 1185 Gln Gln Asp Leu Leu Asp Gly Ala Val Leu Tyr Ser His Asn Gly 1190 1195 1200 Ser Leu Ser Pro Arg Asp Thr Met Ala Phe Ser Val Glu Ala Gly 1205 1210 1215 Pro Val His Thr Asp Ala Thr Leu Gln Val Thr Ile Ala Leu Glu 1220 1225 1230 Gly Pro Leu Ala Pro Leu Lys Leu Val Arg His Lys Lys Ile Tyr 1235 1240 1245 Val Phe Gln Gly Glu Ala Ala Glu Ile Arg Arg Asp Gln Leu Glu 1250 1255 1260 Ala Ala Gln Glu Ala Val Pro Pro Ala Asp Ile Val Phe Ser Val 1265 1270 1275 Lys Ser Pro Pro Ser Ala Gly Tyr Leu Val Met Val Ser Arg Gly 1280 1285 1290 Ala Leu Ala Asp Glu Pro Pro Ser Leu Asp Pro Val Gln Ser Phe 1295 1300 1305 Ser Gln Glu Ala Val Asp Thr Gly Arg Val Leu Tyr Leu His Ser 1310 1315 1320 Arg Pro Glu Ala Trp Ser Asp Ala Phe Ser Leu Asp Val Ala Ser 1325 1330 1335 Gly Leu Gly Ala Pro Leu Glu Gly Val Leu Val Glu Leu Glu Val 1340 1345 1350 Leu Pro Ala Ala Ile Pro Leu Glu Ala Gln Asn Phe Ser Val Pro 1355 1360 1365 Glu Gly Gly Ser Leu Thr Leu Ala Pro Pro Leu Leu Arg Val Ser 1370 1375 1380 Gly Pro Tyr Phe Pro Thr Leu Leu Gly Leu Ser Leu Gln Val Leu 1385 1390 1395 Glu Pro Pro Gln His Gly Ala Leu Gln Lys Glu Asp Gly Pro Gln 1400 1405 1410 Ala Arg Thr Leu Ser Ala Phe Ser Trp Arg Met Val Glu Glu Gln 1415 1420 1425 Leu Ile Arg Tyr Val His Asp Gly Ser Glu Thr Leu Thr Asp Ser 1430 1435 1440 Phe Val Leu Met Ala Asn Ala Ser Glu Met Asp Arg Gln Ser His 1445 1450 1455 Pro Val Ala Phe Thr Val Thr Val Leu Pro Val Asn Asp Gln Pro 1460 1465 1470 Pro Ile Leu Thr Thr Asn Thr Gly Leu Gln Met Trp Glu Gly Ala 1475 1480 1485 Thr Ala Pro Ile Pro Ala Glu Ala Leu Arg Ser Thr Asp Gly Asp 1490 1495 1500 Ser Gly Ser Glu Asp Leu Val Tyr Thr Ile Glu Gln Pro Ser Asn 1505 1510 1515 Gly Arg Val Val Leu Arg Gly Ala Pro Gly Thr Glu Val Arg Ser 1520 1525 1530 Phe Thr Gln Ala Gln Leu Asp Gly Gly Leu Val Leu Phe Ser His 1535 1540 1545 Arg Gly Thr Leu Asp Gly Gly Phe Arg Phe Arg Leu Ser Asp Gly 1550 1555 1560 Glu His Thr Ser Pro Gly His Phe Phe Arg Val Thr Ala Gln Lys 1565 1570 1575 Gln Val Leu Leu Ser Leu Lys Gly Ser Gln Thr Leu Thr Val Cys 1580 1585 1590 Pro Gly Ser Val Gln Pro Leu Ser Ser Gln Thr Leu Arg Ala Ser 1595 1600 1605 Ser Ser Ala Gly Thr Asp Pro Gln Leu Leu Leu Tyr Arg Val Val 1610 1615 1620 Arg Gly Pro Gln Leu Gly Arg Leu Phe His Ala Gln Gln Asp Ser 1625 1630 1635 Thr Gly Glu Ala Leu Val Asn Phe Thr Gln Ala Glu Val Tyr Ala 1640 1645 1650 Gly Asn Ile Leu Tyr Glu His Glu Met Pro Pro Glu Pro Phe Trp 1655 1660 1665 Glu Ala His Asp Thr Leu Glu Leu Gln Leu Ser Ser Pro Pro Ala 1670 1675 1680 Arg Asp Val Ala Ala Thr Leu Ala Val Ala Val Ser Phe Glu Ala 1685 1690 1695 Ala Cys Pro Gln Arg Pro Ser His Leu Trp Lys Asn Lys Gly Leu 1700 1705 1710 Trp Val Pro Glu Gly Gln Arg Ala Arg Ile Thr Val Ala Ala Leu 1715 1720 1725 Asp Ala Ser Asn Leu Leu Ala Ser Val Pro Ser Pro Gln Arg Ser 1730 1735 1740 Glu His Asp Val Leu Phe Gln Val Thr Gln Phe Pro Ser Arg Gly 1745 1750 1755 Gln Leu Leu Val Ser Glu Glu Pro Leu His Ala Gly Gln Pro His 1760 1765 1770 Phe Leu Gln Ser Gln Leu Ala Ala Gly Gln Leu Val Tyr Ala His 1775 1780 1785 Gly Gly Gly Gly Thr Gln Gln Asp Gly Phe His Phe Arg Ala His 1790 1795 1800 Leu Gln Gly Pro Ala Gly Ala Ser Val Ala Gly Pro Gln Thr Ser 1805 1810 1815 Glu Ala Phe Ala Ile Thr Val Arg Asp Val Asn Glu Arg Pro Pro 1820 1825 1830 Gln Pro Gln Ala Ser Val Pro Leu Arg Leu Thr Arg Gly Ser Arg 1835 1840 1845 Ala Pro Ile Ser Arg Ala Gln Leu Ser Val Val Asp Pro Asp Ser 1850 1855 1860 Ala Pro Gly Glu Ile Glu Tyr Glu Val Gln Arg Ala Pro His Asn 1865 1870 1875 Gly Phe Leu Ser Leu Val Gly Gly Gly Leu Gly Pro Val Thr Arg 1880 1885 1890 Phe Thr Gln Ala Asp Val Asp Ser Gly Arg Leu Ala Phe Val Ala 1895 1900 1905 Asn Gly Ser Ser Val Ala Gly Ile Phe Gln Leu Ser Met Ser Asp 1910 1915 1920 Gly Ala Ser Pro Pro Leu Pro Met Ser Leu Ala Val Asp Ile Leu 1925 1930 1935 Pro Ser Ala Ile Glu Val Gln Leu Arg Ala Pro Leu Glu Val Pro 1940 1945 1950 Gln Ala Leu Gly Arg Ser Ser Leu Ser Gln Gln Gln Leu Arg Val 1955 1960 1965 Val Ser Asp Arg Glu Glu Pro Glu Ala Ala Tyr Arg Leu Ile Gln 1970 1975 1980 Gly Pro Gln Tyr Gly His Leu Leu Val Gly Gly Arg Pro Thr Ser 1985 1990 1995 Ala Phe Ser Gln Phe Gln Ile Asp Gln Gly Glu Val Val Phe Ala 2000 2005 2010 Phe Thr Asn Phe Ser Ser Ser His Asp His Phe Arg Val Leu Ala 2015 2020 2025 Leu Ala Arg Gly Val Asn Ala Ser Ala Val Val Asn Val Thr Val 2030 2035 2040 Arg Ala Leu Leu His Val Trp Ala Gly Gly Pro Trp Pro Gln Gly 2045 2050 2055 Ala Thr Leu Arg Leu Asp Pro Thr Val Leu Asp Ala Gly Glu Leu 2060 2065 2070 Ala Asn Arg Thr Gly Ser Val Pro Arg Phe Arg Leu Leu Glu Gly 2075 2080 2085 Pro Arg His Gly Arg Val Val Arg Val Pro Arg Ala Arg Thr Glu 2090 2095 2100 Pro Gly Gly Ser Gln Leu Val Glu Gln Phe Thr Gln Gln Asp Leu 2105 2110 2115 Glu Asp Gly Arg Leu Gly Leu Glu Val Gly Arg Pro Glu Gly Arg 2120 2125 2130 Ala Pro Gly Pro Ala Gly Asp Ser Leu Thr Leu Glu Leu Trp Ala 2135 2140 2145 Gln Gly Val Pro Pro Ala Val Ala Ser Leu Asp Phe Ala Thr Glu 2150 2155 2160 Pro Tyr Asn Ala Ala Arg Pro Tyr Ser Val Ala Leu Leu Ser Val 2165 2170 2175 Pro Glu Ala Ala Arg Thr Glu Ala Gly Lys Pro Glu Ser Ser Thr 2180 2185 2190 Pro Thr Gly Glu Pro Gly Pro Met Ala Ser Ser Pro Glu Pro Ala 2195 2200 2205 Val Ala Lys Gly Gly Phe Leu Ser Phe Leu Glu Ala Asn Met Phe 2210 2215 2220 Ser Val Ile Ile Pro Met Cys Leu Val Leu Leu Leu Leu Ala Leu 2225 2230 2235 Ile Leu Pro Leu Leu Phe Tyr Leu Arg Lys Arg Asn Lys Thr Gly 2240 2245 2250 Lys His Asp Val Gln Val Leu Thr Ala Lys Pro Arg Asn Gly Leu 2255 2260 2265 Ala Gly Asp Thr Glu Thr Phe Arg Lys Val Glu Pro Gly Gln Ala 2270 2275 2280 Ile Pro Leu Thr Ala Val Pro Gly Gln Gly Pro Pro Pro Gly Gly 2285 2290 2295 Gln Pro Asp Pro Glu Leu Leu Gln Phe Cys Arg Thr Pro Asn Pro 2300 2305 2310 Ala Leu Lys Asn Gly Gln Tyr Trp Val 2315 2320 <![CDATA[<210> 94]]> <![CDATA[<211> 1210]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 94]]> Met Arg Pro Ser Gly Thr Ala Gly Ala Ala Leu Leu Ala Leu Leu Ala 1 5 10 15 Ala Leu Cys Pro Ala Ser Arg Ala Leu Glu Glu Lys Lys Val Cys Gln 20 25 30 Gly Thr Ser Asn Lys Leu Thr Gln Leu Gly Thr Phe Glu Asp His Phe 35 40 45 Leu Ser Leu Gln Arg Met Phe Asn Asn Cys Glu Val Val Leu Gly Asn 50 55 60 Leu Glu Ile Thr Tyr Val Gln Arg Asn Tyr Asp Leu Ser Phe Leu Lys 65 70 75 80 Thr Ile Gln Glu Val Ala Gly Tyr Val Leu Ile Ala Leu Asn Thr Val 85 90 95 Glu Arg Ile Pro Leu Glu Asn Leu Gln Ile Ile Arg Gly Asn Met Tyr 100 105 110 Tyr Glu Asn Ser Tyr Ala Leu Ala Val Leu Ser Asn Tyr Asp Ala Asn 115 120 125 Lys Thr Gly Leu Lys Glu Leu Pro Met Arg Asn Leu Gln Glu Ile Leu 130 135 140 His Gly Ala Val Arg Phe Ser Asn Asn Pro Ala Leu Cys Asn Val Glu 145 150 155 160 Ser Ile Gln Trp Arg Asp Ile Val Ser Ser Asp Phe Leu Ser Asn Met 165 170 175 Ser Met Asp Phe Gln Asn His Leu Gly Ser Cys Gln Lys Cys Asp Pro 180 185 190 Ser Cys Pro Asn Gly Ser Cys Trp Gly Ala Gly Glu Glu Asn Cys Gln 195 200 205 Lys Leu Thr Lys Ile Ile Cys Ala Gln Gln Cys Ser Gly Arg Cys Arg 210 215 220 Gly Lys Ser Pro Ser Asp Cys Cys His Asn Gln Cys Ala Ala Gly Cys 225 230 235 240 Thr Gly Pro Arg Glu Ser Asp Cys Leu Val Cys Arg Lys Phe Arg Asp 245 250 255 Glu Ala Thr Cys Lys Asp Thr Cys Pro Pro Leu Met Leu Tyr Asn Pro 260 265 270 Thr Thr Tyr Gln Met Asp Val Asn Pro Glu Gly Lys Tyr Ser Phe Gly 275 280 285 Ala Thr Cys Val Lys Lys Cys Pro Arg Asn Tyr Val Val Thr Asp His 290 295 300 Gly Ser Cys Val Arg Ala Cys Gly Ala Asp Ser Tyr Glu Met Glu Glu 305 310 315 320 Asp Gly Val Arg Lys Cys Lys Lys Cys Glu Gly Pro Cys Arg Lys Val 325 330 335 Cys Asn Gly Ile Gly Ile Gly Glu Phe Lys Asp Ser Leu Ser Ile Asn 340 345 350 Ala Thr Asn Ile Lys His Phe Lys Asn Cys Thr Ser Ile Ser Gly Asp 355 360 365 Leu His Ile Leu Pro Val Ala Phe Arg Gly Asp Ser Phe Thr His Thr 370 375 380 Pro Pro Leu Asp Pro Gln Glu Leu Asp Ile Leu Lys Thr Val Lys Glu 385 390 395 400 Ile Thr Gly Phe Leu Leu Ile Gln Ala Trp Pro Glu Asn Arg Thr Asp 405 410 415 Leu His Ala Phe Glu Asn Leu Glu Ile Ile Arg Gly Arg Thr Lys Gln 420 425 430 His Gly Gln Phe Ser Leu Ala Val Val Ser Leu Asn Ile Thr Ser Leu 435 440 445 Gly Leu Arg Ser Leu Lys Glu Ile Ser Asp Gly Asp Val Ile Ile Ser 450 455 460 Gly Asn Lys Asn Leu Cys Tyr Ala Asn Thr Ile Asn Trp Lys Lys Leu 465 470 475 480 Phe Gly Thr Ser Gly Gln Lys Thr Lys Ile Ile Ser Asn Arg Gly Glu 485 490 495 Asn Ser Cys Lys Ala Thr Gly Gln Val Cys His Ala Leu Cys Ser Pro 500 505 510 Glu Gly Cys Trp Gly Pro Glu Pro Arg Asp Cys Val Ser Cys Arg Asn 515 520 525 Val Ser Arg Gly Arg Glu Cys Val Asp Lys Cys Asn Leu Leu Glu Gly 530 535 540 Glu Pro Arg Glu Phe Val Glu Asn Ser Glu Cys Ile Gln Cys His Pro 545 550 555 560 Glu Cys Leu Pro Gln Ala Met Asn Ile Thr Cys Thr Gly Arg Gly Pro 565 570 575 Asp Asn Cys Ile Gln Cys Ala His Tyr Ile Asp Gly Pro His Cys Val 580 585 590 Lys Thr Cys Pro Ala Gly Val Met Gly Glu Asn Asn Thr Leu Val Trp 595 600 605 Lys Tyr Ala Asp Ala Gly His Val Cys His Leu Cys His Pro Asn Cys 610 615 620 Thr Tyr Gly Cys Thr Gly Pro Gly Leu Glu Gly Cys Pro Thr Asn Gly 625 630 635 640 Pro Lys Ile Pro Ser Ile Ala Thr Gly Met Val Gly Ala Leu Leu Leu 645 650 655 Leu Leu Val Val Ala Leu Gly Ile Gly Leu Phe Met Arg Arg Arg His 660 665 670 Ile Val Arg Lys Arg Thr Leu Arg Arg Leu Leu Gln Glu Arg Glu Leu 675 680 685 Val Glu Pro Leu Thr Pro Ser Gly Glu Ala Pro Asn Gln Ala Leu Leu 690 695 700 Arg Ile Leu Lys Glu Thr Glu Phe Lys Lys Ile Lys Val Leu Gly Ser 705 710 715 720 Gly Ala Phe Gly Thr Val Tyr Lys Gly Leu Trp Ile Pro Glu Gly Glu 725 730 735 Lys Val Lys Ile Pro Val Ala Ile Lys Glu Leu Arg Glu Ala Thr Ser 740 745 750 Pro Lys Ala Asn Lys Glu Ile Leu Asp Glu Ala Tyr Val Met Ala Ser 755 760 765 Val Asp Asn Pro His Val Cys Arg Leu Leu Gly Ile Cys Leu Thr Ser 770 775 780 Thr Val Gln Leu Ile Thr Gln Leu Met Pro Phe Gly Cys Leu Leu Asp 785 790 795 800 Tyr Val Arg Glu His Lys Asp Asn Ile Gly Ser Gln Tyr Leu Leu Asn 805 810 815 Trp Cys Val Gln Ile Ala Lys Gly Met Asn Tyr Leu Glu Asp Arg Arg 820 825 830 Leu Val His Arg Asp Leu Ala Ala Arg Asn Val Leu Val Lys Thr Pro 835 840 845 Gln His Val Lys Ile Thr Asp Phe Gly Leu Ala Lys Leu Leu Gly Ala 850 855 860 Glu Glu Lys Glu Tyr His Ala Glu Gly Gly Lys Val Pro Ile Lys Trp 865 870 875 880 Met Ala Leu Glu Ser Ile Leu His Arg Ile Tyr Thr His Gln Ser Asp 885 890 895 Val Trp Ser Tyr Gly Val Thr Val Trp Glu Leu Met Thr Phe Gly Ser 900 905 910 Lys Pro Tyr Asp Gly Ile Pro Ala Ser Glu Ile Ser Ser Ile Leu Glu 915 920 925 Lys Gly Glu Arg Leu Pro Gln Pro Pro Ile Cys Thr Ile Asp Val Tyr 930 935 940 Met Ile Met Val Lys Cys Trp Met Ile Asp Ala Asp Ser Arg Pro Lys 945 950 955 960 Phe Arg Glu Leu Ile Ile Glu Phe Ser Lys Met Ala Arg Asp Pro Gln 965 970 975 Arg Tyr Leu Val Ile Gln Gly Asp Glu Arg Met His Leu Pro Ser Pro 980 985 990 Thr Asp Ser Asn Phe Tyr Arg Ala Leu Met Asp Glu Glu Asp Met Asp 995 1000 1005 Asp Val Val Asp Ala Asp Glu Tyr Leu Ile Pro Gln Gln Gly Phe 1010 1015 1020 Phe Ser Ser Pro Ser Thr Ser Arg Thr Pro Leu Leu Ser Ser Leu 1025 1030 1035 Ser Ala Thr Ser Asn Asn Ser Thr Val Ala Cys Ile Asp Arg Asn 1040 1045 1050 Gly Leu Gln Ser Cys Pro Ile Lys Glu Asp Ser Phe Leu Gln Arg 1055 1060 1065 Tyr Ser Ser Asp Pro Thr Gly Ala Leu Thr Glu Asp Ser Ile Asp 1070 1075 1080 Asp Thr Phe Leu Pro Val Pro Glu Tyr Ile Asn Gln Ser Val Pro 1085 1090 1095 Lys Arg Pro Ala Gly Ser Val Gln Asn Pro Val Tyr His Asn Gln 1100 1105 1110 Pro Leu Asn Pro Ala Pro Ser Arg Asp Pro His Tyr Gln Asp Pro 1115 1120 1125 His Ser Thr Ala Val Gly Asn Pro Glu Tyr Leu Asn Thr Val Gln 1130 1135 1140 Pro Thr Cys Val Asn Ser Thr Phe Asp Ser Pro Ala His Trp Ala 1145 1150 1155 Gln Lys Gly Ser His Gln Ile Ser Leu Asp Asn Pro Asp Tyr Gln 1160 1165 1170 Gln Asp Phe Phe Pro Lys Glu Ala Lys Pro Asn Gly Ile Phe Lys 1175 1180 1185 Gly Ser Thr Ala Glu Asn Ala Glu Tyr Leu Arg Val Ala Pro Gln 1190 1195 1200 Ser Ser Glu Phe Ile Gly Ala 1205 1210 <![CDATA[<210> 95]]> <![CDATA[<211> 556]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 95]]> Met Pro Pro Pro Arg Leu Leu Phe Phe Leu Leu Phe Leu Thr Pro Met 1 5 10 15 Glu Val Arg Pro Glu Glu Pro Leu Val Val Lys Val Glu Glu Gly Asp 20 25 30 Asn Ala Val Leu Gln Cys Leu Lys Gly Thr Ser Asp Gly Pro Thr Gln 35 40 45 Gln Leu Thr Trp Ser Arg Glu Ser Pro Leu Lys Pro Phe Leu Lys Leu 50 55 60 Ser Leu Gly Leu Pro Gly Leu Gly Ile His Met Arg Pro Leu Ala Ile 65 70 75 80 Trp Leu Phe Ile Phe Asn Val Ser Gln Gln Met Gly Gly Phe Tyr Leu 85 90 95 Cys Gln Pro Gly Pro Pro Ser Glu Lys Ala Trp Gln Pro Gly Trp Thr 100 105 110 Val Asn Val Glu Gly Ser Gly Glu Leu Phe Arg Trp Asn Val Ser Asp 115 120 125 Leu Gly Gly Leu Gly Cys Gly Leu Lys Asn Arg Ser Ser Glu Gly Pro 130 135 140 Ser Ser Pro Ser Gly Lys Leu Met Ser Pro Lys Leu Tyr Val Trp Ala 145 150 155 160 Lys Asp Arg Pro Glu Ile Trp Glu Gly Glu Pro Pro Cys Leu Pro Pro 165 170 175 Arg Asp Ser Leu Asn Gln Ser Leu Ser Gln Asp Leu Thr Met Ala Pro 180 185 190 Gly Ser Thr Leu Trp Leu Ser Cys Gly Val Pro Pro Asp Ser Val Ser 195 200 205 Arg Gly Pro Leu Ser Trp Thr His Val His Pro Lys Gly Pro Lys Ser 210 215 220 Leu Leu Ser Leu Glu Leu Lys Asp Asp Arg Pro Ala Arg Asp Met Trp 225 230 235 240 Val Met Glu Thr Gly Leu Leu Leu Pro Arg Ala Thr Ala Gln Asp Ala 245 250 255 Gly Lys Tyr Tyr Cys His Arg Gly Asn Leu Thr Met Ser Phe His Leu 260 265 270 Glu Ile Thr Ala Arg Pro Val Leu Trp His Trp Leu Leu Arg Thr Gly 275 280 285 Gly Trp Lys Val Ser Ala Val Thr Leu Ala Tyr Leu Ile Phe Cys Leu 290 295 300 Cys Ser Leu Val Gly Ile Leu His Leu Gln Arg Ala Leu Val Leu Arg 305 310 315 320 Arg Lys Arg Lys Arg Met Thr Asp Pro Thr Arg Arg Phe Phe Lys Val 325 330 335 Thr Pro Pro Pro Gly Ser Gly Pro Gln Asn Gln Tyr Gly Asn Val Leu 340 345 350 Ser Leu Pro Thr Pro Thr Ser Gly Leu Gly Arg Ala Gln Arg Trp Ala 355 360 365 Ala Gly Leu Gly Gly Thr Ala Pro Ser Tyr Gly Asn Pro Ser Ser Asp 370 375 380 Val Gln Ala Asp Gly Ala Leu Gly Ser Arg Ser Pro Pro Gly Val Gly 385 390 395 400 Pro Glu Glu Glu Glu Gly Glu Gly Tyr Glu Glu Pro Asp Ser Glu Glu 405 410 415 Asp Ser Glu Phe Tyr Glu Asn Asp Ser Asn Leu Gly Gln Asp Gln Leu 420 425 430 Ser Gln Asp Gly Ser Gly Tyr Glu Asn Pro Glu Asp Glu Pro Leu Gly 435 440 445 Pro Glu Asp Glu Asp Ser Phe Ser Asn Ala Glu Ser Tyr Glu Asn Glu 450 455 460 Asp Glu Glu Leu Thr Gln Pro Val Ala Arg Thr Met Asp Phe Leu Ser 465 470 475 480 Pro His Gly Ser Ala Trp Asp Pro Ser Arg Glu Ala Thr Ser Leu Gly 485 490 495 Ser Gln Ser Tyr Glu Asp Met Arg Gly Ile Leu Tyr Ala Ala Pro Gln 500 505 510 Leu Arg Ser Ile Arg Gly Gln Pro Gly Pro Asn His Glu Glu Asp Ala 515 520 525 Asp Ser Tyr Glu Asn Met Asp Asn Pro Asp Gly Pro Asp Pro Ala Trp 530 535 540 Gly Gly Gly Gly Arg Met Gly Thr Trp Ser Thr Arg 545 550 555 <![CDATA[<210> 96]]> <![CDATA[<211> 297]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 96]]> Met Thr Thr Pro Arg Asn Ser Val Asn Gly Thr Phe Pro Ala Glu Pro 1 5 10 15 Met Lys Gly Pro Ile Ala Met Gln Ser Gly Pro Lys Pro Leu Phe Arg 20 25 30 Arg Met Ser Ser Leu Val Gly Pro Thr Gln Ser Phe Phe Met Arg Glu 35 40 45 Ser Lys Thr Leu Gly Ala Val Gln Ile Met Asn Gly Leu Phe His Ile 50 55 60 Ala Leu Gly Gly Leu Leu Met Ile Pro Ala Gly Ile Tyr Ala Pro Ile 65 70 75 80 Cys Val Thr Val Trp Tyr Pro Leu Trp Gly Gly Ile Met Tyr Ile Ile 85 90 95 Ser Gly Ser Leu Leu Ala Ala Thr Glu Lys Asn Ser Arg Lys Cys Leu 100 105 110 Val Lys Gly Lys Met Ile Met Asn Ser Leu Ser Leu Phe Ala Ala Ile 115 120 125 Ser Gly Met Ile Leu Ser Ile Met Asp Ile Leu Asn Ile Lys Ile Ser 130 135 140 His Phe Leu Lys Met Glu Ser Leu Asn Phe Ile Arg Ala His Thr Pro 145 150 155 160 Tyr Ile Asn Ile Tyr Asn Cys Glu Pro Ala Asn Pro Ser Glu Lys Asn 165 170 175 Ser Pro Ser Thr Gln Tyr Cys Tyr Ser Ile Gln Ser Leu Phe Leu Gly 180 185 190 Ile Leu Ser Val Met Leu Ile Phe Ala Phe Phe Gln Glu Leu Val Ile 195 200 205 Ala Gly Ile Val Glu Asn Glu Trp Lys Arg Thr Cys Ser Arg Pro Lys 210 215 220 Ser Asn Ile Val Leu Leu Ser Ala Glu Glu Lys Lys Glu Gln Thr Ile 225 230 235 240 Glu Ile Lys Glu Glu Val Val Gly Leu Thr Glu Thr Ser Ser Gln Pro 245 250 255 Lys Asn Glu Glu Asp Ile Glu Ile Ile Pro Ile Gln Glu Glu Glu Glu 260 265 270 Glu Glu Thr Glu Thr Asn Phe Pro Glu Pro Pro Gln Asp Gln Glu Ser 275 280 285 Ser Pro Ile Glu Asn Asp Ser Ser Pro 290 295 <![CDATA[<210> 97]]> <![CDATA[<211> 364]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 97]]> Met Pro Leu Leu Leu Leu Leu Pro Leu Leu Trp Ala Gly Ala Leu Ala 1 5 10 15 Met Asp Pro Asn Phe Trp Leu Gln Val Gln Glu Ser Val Thr Val Gln 20 25 30 Glu Gly Leu Cys Val Leu Val Pro Cys Thr Phe Phe His Pro Ile Pro 35 40 45 Tyr Tyr Asp Lys Asn Ser Pro Val His Gly Tyr Trp Phe Arg Glu Gly 50 55 60 Ala Ile Ile Ser Arg Asp Ser Pro Val Ala Thr Asn Lys Leu Asp Gln 65 70 75 80 Glu Val Gln Glu Glu Thr Gln Gly Arg Phe Arg Leu Leu Gly Asp Pro 85 90 95 Ser Arg Asn Asn Cys Ser Leu Ser Ile Val Asp Ala Arg Arg Arg Asp 100 105 110 Asn Gly Ser Tyr Phe Phe Arg Met Glu Arg Gly Ser Thr Lys Tyr Ser 115 120 125 Tyr Lys Ser Pro Gln Leu Ser Val His Val Thr Asp Leu Thr His Arg 130 135 140 Pro Lys Ile Leu Ile Pro Gly Thr Leu Glu Pro Gly His Ser Lys Asn 145 150 155 160 Leu Thr Cys Ser Val Ser Trp Ala Cys Glu Gln Gly Thr Pro Pro Ile 165 170 175 Phe Ser Trp Leu Ser Ala Ala Pro Thr Ser Leu Gly Pro Arg Thr Thr 180 185 190 His Ser Ser Val Leu Ile Ile Thr Pro Arg Pro Gln Asp His Gly Thr 195 200 205 Asn Leu Thr Cys Gln Val Lys Phe Ala Gly Ala Gly Val Thr Thr Glu 210 215 220 Arg Thr Ile Gln Leu Asn Val Thr Tyr Val Pro Gln Asn Pro Thr Thr 225 230 235 240 Gly Ile Phe Pro Gly Asp Gly Ser Gly Lys Gln Glu Thr Arg Ala Gly 245 250 255 Val Val His Gly Ala Ile Gly Gly Ala Gly Val Thr Ala Leu Leu Ala 260 265 270 Leu Cys Leu Cys Leu Ile Phe Phe Ile Val Lys Thr His Arg Arg Lys 275 280 285 Ala Ala Arg Thr Ala Val Gly Arg Asn Asp Thr His Pro Thr Thr Gly 290 295 300 Ser Ala Ser Pro Lys His Gln Lys Lys Ser Lys Leu His Gly Pro Thr 305 310 315 320 Glu Thr Ser Ser Cys Ser Gly Ala Ala Pro Thr Val Glu Met Asp Glu 325 330 335 Glu Leu His Tyr Ala Ser Leu Asn Phe His Gly Met Asn Pro Ser Lys 340 345 350 Asp Thr Ser Thr Glu Tyr Ser Glu Val Arg Thr Gln 355 360 <![CDATA[<210> 98]]> <![CDATA[<211> 277]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 98]]> Met Cys Val Gly Ala Arg Arg Leu Gly Arg Gly Pro Cys Ala Ala Leu 1 5 10 15 Leu Leu Leu Gly Leu Gly Leu Ser Thr Val Thr Gly Leu His Cys Val 20 25 30 Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys His Glu Cys Arg Pro 35 40 45 Gly Asn Gly Met Val Ser Arg Cys Ser Arg Ser Gln Asn Thr Val Cys 50 55 60 Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val Ser Ser Lys Pro 65 70 75 80 Cys Lys Pro Cys Thr Trp Cys Asn Leu Arg Ser Gly Ser Glu Arg Lys 85 90 95 Gln Leu Cys Thr Ala Thr Gln Asp Thr Val Cys Arg Cys Arg Ala Gly 100 105 110 Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp Cys Ala Pro Cys 115 120 125 Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala Cys Lys Pro Trp 130 135 140 Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln Pro Ala Ser Asn 145 150 155 160 Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro Ala Thr Gln Pro 165 170 175 Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Ile Thr Val Gln Pro Thr 180 185 190 Glu Ala Trp Pro Arg Thr Ser Gln Gly Pro Ser Thr Arg Pro Val Glu 195 200 205 Val Pro Gly Gly Arg Ala Val Ala Ala Ile Leu Gly Leu Gly Leu Val 210 215 220 Leu Gly Leu Leu Gly Pro Leu Ala Ile Leu Leu Ala Leu Tyr Leu Leu 225 230 235 240 Arg Arg Asp Gln Arg Leu Pro Pro Asp Ala His Lys Pro Pro Gly Gly 245 250 255 Gly Ser Phe Arg Thr Pro Ile Gln Glu Glu Gln Ala Asp Ala His Ser 260 265 270 Thr Leu Ala Lys Ile 275 <![CDATA[<210> 99]]> <![CDATA[<211> 255]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 99]]> Met Gly Asn Ser Cys Tyr Asn Ile Val Ala Thr Leu Leu Leu Val Leu 1 5 10 15 Asn Phe Glu Arg Thr Arg Ser Leu Gln Asp Pro Cys Ser Asn Cys Pro 20 25 30 Ala Gly Thr Phe Cys Asp Asn Asn Arg Asn Gln Ile Cys Ser Pro Cys 35 40 45 Pro Pro Asn Ser Phe Ser Ser Ala Gly Gly Gln Arg Thr Cys Asp Ile 50 55 60 Cys Arg Gln Cys Lys Gly Val Phe Arg Thr Arg Lys Glu Cys Ser Ser 65 70 75 80 Thr Ser Asn Ala Glu Cys Asp Cys Thr Pro Gly Phe His Cys Leu Gly 85 90 95 Ala Gly Cys Ser Met Cys Glu Gln Asp Cys Lys Gln Gly Gln Glu Leu 100 105 110 Thr Lys Lys Gly Cys Lys Asp Cys Cys Phe Gly Thr Phe Asn Asp Gln 115 120 125 Lys Arg Gly Ile Cys Arg Pro Trp Thr Asn Cys Ser Leu Asp Gly Lys 130 135 140 Ser Val Leu Val Asn Gly Thr Lys Glu Arg Asp Val Val Cys Gly Pro 145 150 155 160 Ser Pro Ala Asp Leu Ser Pro Gly Ala Ser Ser Val Thr Pro Pro Ala 165 170 175 Pro Ala Arg Glu Pro Gly His Ser Pro Gln Ile Ile Ser Phe Phe Leu 180 185 190 Ala Leu Thr Ser Thr Ala Leu Leu Phe Leu Leu Phe Phe Leu Thr Leu 195 200 205 Arg Phe Ser Val Val Lys Arg Gly Arg Lys Lys Leu Leu Tyr Ile Phe 210 215 220 Lys Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly 225 230 235 240 Cys Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 245 250 255 <![CDATA[<210> 100]]> <![CDATA[<211> 260]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 100]]> Met Ala Arg Pro His Pro Trp Trp Leu Cys Val Leu Gly Thr Leu Val 1 5 10 15 Gly Leu Ser Ala Thr Pro Ala Pro Lys Ser Cys Pro Glu Arg His Tyr 20 25 30 Trp Ala Gln Gly Lys Leu Cys Cys Gln Met Cys Glu Pro Gly Thr Phe 35 40 45 Leu Val Lys Asp Cys Asp Gln His Arg Lys Ala Ala Gln Cys Asp Pro 50 55 60 Cys Ile Pro Gly Val Ser Phe Ser Pro Asp His His Thr Arg Pro His 65 70 75 80 Cys Glu Ser Cys Arg His Cys Asn Ser Gly Leu Leu Val Arg Asn Cys 85 90 95 Thr Ile Thr Ala Asn Ala Glu Cys Ala Cys Arg Asn Gly Trp Gln Cys 100 105 110 Arg Asp Lys Glu Cys Thr Glu Cys Asp Pro Leu Pro Asn Pro Ser Leu 115 120 125 Thr Ala Arg Ser Ser Gln Ala Leu Ser Pro His Pro Gln Pro Thr His 130 135 140 Leu Pro Tyr Val Ser Glu Met Leu Glu Ala Arg Thr Ala Gly His Met 145 150 155 160 Gln Thr Leu Ala Asp Phe Arg Gln Leu Pro Ala Arg Thr Leu Ser Thr 165 170 175 His Trp Pro Pro Gln Arg Ser Leu Cys Ser Ser Asp Phe Ile Arg Ile 180 185 190 Leu Val Ile Phe Ser Gly Met Phe Leu Val Phe Thr Leu Ala Gly Ala 195 200 205 Leu Phe Leu His Gln Arg Arg Lys Tyr Arg Ser Asn Lys Gly Glu Ser 210 215 220 Pro Val Glu Pro Ala Glu Pro Cys His Tyr Ser Cys Pro Arg Glu Glu 225 230 235 240 Glu Gly Ser Thr Ile Pro Ile Gln Glu Asp Tyr Arg Lys Pro Glu Pro 245 250 255 Ala Cys Ser Pro 260 <![CDATA[<210> 101]]> <![CDATA[<211> 283]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 101]]> Met Glu Pro Pro Gly Asp Trp Gly Pro Pro Pro Trp Arg Ser Thr Pro 1 5 10 15 Lys Thr Asp Val Leu Arg Leu Val Leu Tyr Leu Thr Phe Leu Gly Ala 20 25 30 Pro Cys Tyr Ala Pro Ala Leu Pro Ser Cys Lys Glu Asp Glu Tyr Pro 35 40 45 Val Gly Ser Glu Cys Cys Pro Lys Cys Ser Pro Gly Tyr Arg Val Lys 50 55 60 Glu Ala Cys Gly Glu Leu Thr Gly Thr Val Cys Glu Pro Cys Pro Pro 65 70 75 80 Gly Thr Tyr Ile Ala His Leu Asn Gly Leu Ser Lys Cys Leu Gln Cys 85 90 95 Gln Met Cys Asp Pro Ala Met Gly Leu Arg Ala Ser Arg Asn Cys Ser 100 105 110 Arg Thr Glu Asn Ala Val Cys Gly Cys Ser Pro Gly His Phe Cys Ile 115 120 125 Val Gln Asp Gly Asp His Cys Ala Ala Cys Arg Ala Tyr Ala Thr Ser 130 135 140 Ser Pro Gly Gln Arg Val Gln Lys Gly Gly Thr Glu Ser Gln Asp Thr 145 150 155 160 Leu Cys Gln Asn Cys Pro Pro Gly Thr Phe Ser Pro Asn Gly Thr Leu 165 170 175 Glu Glu Cys Gln His Gln Thr Lys Cys Ser Trp Leu Val Thr Lys Ala 180 185 190 Gly Ala Gly Thr Ser Ser Ser His Trp Val Trp Trp Phe Leu Ser Gly 195 200 205 Ser Leu Val Ile Val Ile Val Cys Ser Thr Val Gly Leu Ile Ile Cys 210 215 220 Val Lys Arg Arg Lys Pro Arg Gly Asp Val Val Lys Val Ile Val Ser 225 230 235 240 Val Gln Arg Lys Arg Gln Glu Ala Glu Gly Glu Ala Thr Val Ile Glu 245 250 255 Ala Leu Gln Ala Pro Pro Asp Val Thr Thr Val Ala Val Glu Glu Thr 260 265 270 Ile Pro Ser Phe Thr Gly Arg Ser Pro Asn His 275 280 <![CDATA[<210> 102]]> <![CDATA[<211> 595]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 102]]> Met Arg Val Leu Leu Ala Ala Leu Gly Leu Leu Phe Leu Gly Ala Leu 1 5 10 15 Arg Ala Phe Pro Gln Asp Arg Pro Phe Glu Asp Thr Cys His Gly Asn 20 25 30 Pro Ser His Tyr Tyr Asp Lys Ala Val Arg Arg Cys Cys Tyr Arg Cys 35 40 45 Pro Met Gly Leu Phe Pro Thr Gln Gln Cys Pro Gln Arg Pro Thr Asp 50 55 60 Cys Arg Lys Gln Cys Glu Pro Asp Tyr Tyr Leu Asp Glu Ala Asp Arg 65 70 75 80 Cys Thr Ala Cys Val Thr Cys Ser Arg Asp Asp Leu Val Glu Lys Thr 85 90 95 Pro Cys Ala Trp Asn Ser Ser Arg Val Cys Glu Cys Arg Pro Gly Met 100 105 110 Phe Cys Ser Thr Ser Ala Val Asn Ser Cys Ala Arg Cys Phe Phe His 115 120 125 Ser Val Cys Pro Ala Gly Met Ile Val Lys Phe Pro Gly Thr Ala Gln 130 135 140 Lys Asn Thr Val Cys Glu Pro Ala Ser Pro Gly Val Ser Pro Ala Cys 145 150 155 160 Ala Ser Pro Glu Asn Cys Lys Glu Pro Ser Ser Gly Thr Ile Pro Gln 165 170 175 Ala Lys Pro Thr Pro Val Ser Pro Ala Thr Ser Ser Ala Ser Thr Met 180 185 190 Pro Val Arg Gly Gly Thr Arg Leu Ala Gln Glu Ala Ala Ser Lys Leu 195 200 205 Thr Arg Ala Pro Asp Ser Pro Ser Ser Val Gly Arg Pro Ser Ser Asp 210 215 220 Pro Gly Leu Ser Pro Thr Gln Pro Cys Pro Glu Gly Ser Gly Asp Cys 225 230 235 240 Arg Lys Gln Cys Glu Pro Asp Tyr Tyr Leu Asp Glu Ala Gly Arg Cys 245 250 255 Thr Ala Cys Val Ser Cys Ser Arg Asp Asp Leu Val Glu Lys Thr Pro 260 265 270 Cys Ala Trp Asn Ser Ser Arg Thr Cys Glu Cys Arg Pro Gly Met Ile 275 280 285 Cys Ala Thr Ser Ala Thr Asn Ser Cys Ala Arg Cys Val Pro Tyr Pro 290 295 300 Ile Cys Ala Ala Glu Thr Val Thr Lys Pro Gln Asp Met Ala Glu Lys 305 310 315 320 Asp Thr Thr Phe Glu Ala Pro Pro Leu Gly Thr Gln Pro Asp Cys Asn 325 330 335 Pro Thr Pro Glu Asn Gly Glu Ala Pro Ala Ser Thr Ser Pro Thr Gln 340 345 350 Ser Leu Leu Val Asp Ser Gln Ala Ser Lys Thr Leu Pro Ile Pro Thr 355 360 365 Ser Ala Pro Val Ala Leu Ser Ser Thr Gly Lys Pro Val Leu Asp Ala 370 375 380 Gly Pro Val Leu Phe Trp Val Ile Leu Val Leu Val Val Val Val Gly 385 390 395 400 Ser Ser Ala Phe Leu Leu Cys His Arg Arg Ala Cys Arg Lys Arg Ile 405 410 415 Arg Gln Lys Leu His Leu Cys Tyr Pro Val Gln Thr Ser Gln Pro Lys 420 425 430 Leu Glu Leu Val Asp Ser Arg Pro Arg Arg Ser Ser Thr Gln Leu Arg 435 440 445 Ser Gly Ala Ser Val Thr Glu Pro Val Ala Glu Glu Arg Gly Leu Met 450 455 460 Ser Gln Pro Leu Met Glu Thr Cys His Ser Val Gly Ala Ala Tyr Leu 465 470 475 480 Glu Ser Leu Pro Leu Gln Asp Ala Ser Pro Ala Gly Gly Pro Ser Ser 485 490 495 Pro Arg Asp Leu Pro Glu Pro Arg Val Ser Thr Glu His Thr Asn Asn 500 505 510 Lys Ile Glu Lys Ile Tyr Ile Met Lys Ala Asp Thr Val Ile Val Gly 515 520 525 Thr Val Lys Ala Glu Leu Pro Glu Gly Arg Gly Leu Ala Gly Pro Ala 530 535 540 Glu Pro Glu Leu Glu Glu Glu Leu Glu Ala Asp His Thr Pro His Tyr 545 550 555 560 Pro Glu Gln Glu Thr Glu Pro Pro Leu Gly Ser Cys Ser Asp Val Met 565 570 575 Leu Ser Val Glu Glu Glu Gly Lys Glu Asp Pro Leu Pro Thr Ala Ala 580 585 590 Ser Gly Lys 595 <![CDATA[<210> 103]]> <![CDATA[<211> 241]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 智人]]> <![CDATA[<400> 103]]> Met Ala Gln His Gly Ala Met Gly Ala Phe Arg Ala Leu Cys Gly Leu 1 5 10 15 Ala Leu Leu Cys Ala Leu Ser Leu Gly Gln Arg Pro Thr Gly Gly Pro 20 25 30 Gly Cys Gly Pro Gly Arg Leu Leu Leu Gly Thr Gly Thr Asp Ala Arg 35 40 45 Cys Cys Arg Val His Thr Thr Arg Cys Cys Arg Asp Tyr Pro Gly Glu 50 55 60 Glu Cys Cys Ser Glu Trp Asp Cys Met Cys Val Gln Pro Glu Phe His 65 70 75 80 Cys Gly Asp Pro Cys Cys Thr Thr Cys Arg His His Pro Cys Pro Pro 85 90 95 Gly Gln Gly Val Gln Ser Gln Gly Lys Phe Ser Phe Gly Phe Gln Cys 100 105 110 Ile Asp Cys Ala Ser Gly Thr Phe Ser Gly Gly His Glu Gly His Cys 115 120 125 Lys Pro Trp Thr Asp Cys Thr Gln Phe Gly Phe Leu Thr Val Phe Pro 130 135 140 Gly Asn Lys Thr His Asn Ala Val Cys Val Pro Gly Ser Pro Pro Ala 145 150 155 160 Glu Pro Leu Gly Trp Leu Thr Val Val Leu Leu Ala Val Ala Ala Cys 165 170 175 Val Leu Leu Leu Thr Ser Ala Gln Leu Gly Leu His Ile Trp Gln Leu 180 185 190 Arg Ser Gln Cys Met Trp Pro Arg Glu Thr Gln Leu Leu Leu Glu Val 195 200 205 Pro Pro Ser Thr Glu Asp Ala Arg Ser Cys Gln Phe Pro Glu Glu Glu 210 215 220 Arg Gly Glu Arg Ser Ala Glu Glu Lys Gly Arg Leu Gly Asp Leu Trp 225 230 235 240 Val <![CDATA[<210> 104]]> <![CDATA[<211> 272]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 小家鼠]]> <![CDATA[<400> 104]]> Met Tyr Val Trp Val Gln Gln Pro Thr Ala Leu Leu Leu Leu Ala Leu 1 5 10 15 Thr Leu Gly Val Thr Ala Arg Arg Leu Asn Cys Val Lys His Thr Tyr 20 25 30 Pro Ser Gly His Lys Cys Cys Arg Glu Cys Gln Pro Gly His Gly Met 35 40 45 Val Ser Arg Cys Asp His Thr Arg Asp Thr Leu Cys His Pro Cys Glu 50 55 60 Thr Gly Phe Tyr Asn Glu Ala Val Asn Tyr Asp Thr Cys Lys Gln Cys 65 70 75 80 Thr Gln Cys Asn His Arg Ser Gly Ser Glu Leu Lys Gln Asn Cys Thr 85 90 95 Pro Thr Gln Asp Thr Val Cys Arg Cys Arg Pro Gly Thr Gln Pro Arg 100 105 110 Gln Asp Ser Gly Tyr Lys Leu Gly Val Asp Cys Val Pro Cys Pro Pro 115 120 125 Gly His Phe Ser Pro Gly Asn Asn Gln Ala Cys Lys Pro Trp Thr Asn 130 135 140 Cys Thr Leu Ser Gly Lys Gln Thr Arg His Pro Ala Ser Asp Ser Leu 145 150 155 160 Asp Ala Val Cys Glu Asp Arg Ser Leu Leu Ala Thr Leu Leu Trp Glu 165 170 175 Thr Gln Arg Pro Thr Phe Arg Pro Thr Thr Val Gln Ser Thr Thr Val 180 185 190 Trp Pro Arg Thr Ser Glu Leu Pro Ser Pro Pro Thr Leu Val Thr Pro 195 200 205 Glu Gly Pro Ala Phe Ala Val Leu Leu Gly Leu Gly Leu Gly Leu Leu 210 215 220 Ala Pro Leu Thr Val Leu Leu Ala Leu Tyr Leu Leu Arg Lys Ala Trp 225 230 235 240 Arg Leu Pro Asn Thr Pro Lys Pro Cys Trp Gly Asn Ser Phe Arg Thr 245 250 255 Pro Ile Gln Glu Glu His Thr Asp Ala His Phe Thr Leu Ala Lys Ile 260 265 270 <![CDATA[<210> 105]]> <![CDATA[<211> 256]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 小家鼠]]> <![CDATA[<400> 105]]> Met Gly Asn Asn Cys Tyr Asn Val Val Val Ile Val Leu Leu Leu Val 1 5 10 15 Gly Cys Glu Lys Val Gly Ala Val Gln Asn Ser Cys Asp Asn Cys Gln 20 25 30 Pro Gly Thr Phe Cys Arg Lys Tyr Asn Pro Val Cys Lys Ser Cys Pro 35 40 45 Pro Ser Thr Phe Ser Ser Ile Gly Gly Gln Pro Asn Cys Asn Ile Cys 50 55 60 Arg Val Cys Ala Gly Tyr Phe Arg Phe Lys Lys Phe Cys Ser Ser Thr 65 70 75 80 His Asn Ala Glu Cys Glu Cys Ile Glu Gly Phe His Cys Leu Gly Pro 85 90 95 Gln Cys Thr Arg Cys Glu Lys Asp Cys Arg Pro Gly Gln Glu Leu Thr 100 105 110 Lys Gln Gly Cys Lys Thr Cys Ser Leu Gly Thr Phe Asn Asp Gln Asn 115 120 125 Gly Thr Gly Val Cys Arg Pro Trp Thr Asn Cys Ser Leu Asp Gly Arg 130 135 140 Ser Val Leu Lys Thr Gly Thr Thr Glu Lys Asp Val Val Cys Gly Pro 145 150 155 160 Pro Val Val Ser Phe Ser Pro Ser Thr Thr Ile Ser Val Thr Pro Glu 165 170 175 Gly Gly Pro Gly Gly His Ser Leu Gln Val Leu Thr Leu Phe Leu Ala 180 185 190 Leu Thr Ser Ala Leu Leu Leu Ala Leu Ile Phe Ile Thr Leu Leu Phe 195 200 205 Ser Val Leu Lys Trp Ile Arg Lys Lys Phe Pro His Ile Phe Lys Gln 210 215 220 Pro Phe Lys Lys Thr Thr Gly Ala Ala Gln Glu Glu Asp Ala Cys Ser 225 230 235 240 Cys Arg Cys Pro Gln Glu Glu Glu Gly Gly Gly Gly Gly Tyr Glu Leu 245 250 255 <![CDATA[<210> 106]]> <![CDATA[<211> 254]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 獼猴]]> <![CDATA[<400> 106]]> Met Gly Asn Ser Cys Tyr Asn Ile Val Ala Thr Leu Leu Leu Val Leu 1 5 10 15 Asn Phe Glu Arg Thr Arg Ser Leu Gln Asp Leu Cys Ser Asn Cys Pro 20 25 30 Ala Gly Thr Phe Cys Asp Asn Asn Arg Ser Gln Ile Cys Ser Pro Cys 35 40 45 Pro Pro Asn Ser Phe Ser Ser Ala Gly Gly Gln Arg Thr Cys Asp Ile 50 55 60 Cys Arg Gln Cys Lys Gly Val Phe Lys Thr Arg Lys Glu Cys Ser Ser 65 70 75 80 Thr Ser Asn Ala Glu Cys Asp Cys Ile Ser Gly Tyr His Cys Leu Gly 85 90 95 Ala Glu Cys Ser Met Cys Glu Gln Asp Cys Lys Gln Gly Gln Glu Leu 100 105 110 Thr Lys Lys Gly Cys Lys Asp Cys Cys Phe Gly Thr Phe Asn Asp Gln 115 120 125 Lys Arg Gly Ile Cys Arg Pro Trp Thr Asn Cys Ser Leu Asp Gly Lys 130 135 140 Ser Val Leu Val Asn Gly Thr Lys Glu Arg Asp Val Val Cys Gly Pro 145 150 155 160 Ser Pro Ala Asp Leu Ser Pro Gly Ala Ser Ser Ala Thr Pro Pro Ala 165 170 175 Pro Ala Arg Glu Pro Gly His Ser Pro Gln Ile Ile Phe Phe Leu Ala 180 185 190 Leu Thr Ser Thr Val Val Leu Phe Leu Leu Phe Phe Leu Val Leu Arg 195 200 205 Phe Ser Val Val Lys Arg Ser Arg Lys Lys Leu Leu Tyr Ile Phe Lys 210 215 220 Gln Pro Phe Met Arg Pro Val Gln Thr Thr Gln Glu Glu Asp Gly Cys 225 230 235 240 Ser Cys Arg Phe Pro Glu Glu Glu Glu Gly Gly Cys Glu Leu 245 250 <![CDATA[<210> 107]]> <![CDATA[<211> 5]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 107]]> Gly Gly Gly Gly Ser 1 5 <![CDATA[<210> 108]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 108]]> Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 <![CDATA[<210> 109]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 109]]> Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 1 5 10 <![CDATA[<210> 110]]> <![CDATA[<211> 14]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 110]]> Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 1 5 10 <![CDATA[<210> 111]]> <![CDATA[<211> 10]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 111]]> Gly Ser Pro Gly Ser Ser Ser Ser Gly Ser 1 5 10 <![CDATA[<210> 112]]> <![CDATA[<211> 15]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 112]]> Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 1 5 10 15 <![CDATA[<210> 113]]> <![CDATA[<211> 20]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 113]]> Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 1 5 10 15 Gly Gly Gly Ser 20 <![CDATA[<210> 114]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 114]]> Gly Ser Gly Ser Gly Ser Gly Ser 1 5 <![CDATA[<210> 115]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 115]]> Gly Ser Gly Ser Gly Asn Gly Ser 1 5 <![CDATA[<210> 116]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 116]]> Gly Gly Ser Gly Ser Gly Ser Gly 1 5 <![CDATA[<210> 117]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 117]]> Gly Gly Ser Gly Ser Gly 1 5 <![CDATA[<210> 118]]> <![CDATA[<211> 4]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 118]]> Gly Gly Ser Gly 1 <![CDATA[<210> 119]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 119]]> Gly Gly Ser Gly Asn Gly Ser Gly 1 5 <![CDATA[<210> 120]]> <![CDATA[<211> 8]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 120]]> Gly Gly Asn Gly Ser Gly Ser Gly 1 5 <![CDATA[<210> 121]]> <![CDATA[<211> 6]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 肽連接子]]> <![CDATA[<400> 121]]> Gly Gly Asn Gly Ser Gly 1 5 <![CDATA[<210> 122]]> <![CDATA[<211> 186]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 獼猴]]> <![CDATA[<400> 122]]> Leu His Cys Val Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys Gln 1 5 10 15 Glu Cys Arg Pro Gly Asn Gly Met Val Ser Arg Cys Asn Arg Ser Gln 20 25 30 Asn Thr Val Cys Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val 35 40 45 Ser Ala Lys Pro Cys Lys Ala Cys Thr Trp Cys Asn Leu Arg Ser Gly 50 55 60 Ser Glu Arg Lys Gln Pro Cys Thr Ala Thr Gln Asp Thr Val Cys Arg 65 70 75 80 Cys Arg Ala Gly Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp 85 90 95 Cys Ala Pro Cys Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala 100 105 110 Cys Lys Pro Trp Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln 115 120 125 Pro Ala Ser Asn Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro 130 135 140 Pro Thr Gln Pro Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Thr Thr 145 150 155 160 Val Gln Pro Thr Glu Ala Trp Pro Arg Thr Ser Gln Arg Pro Ser Thr 165 170 175 Arg Pro Val Glu Val Pro Arg Gly Pro Ala 180 185 <![CDATA[<210> 123]]> <![CDATA[<211> 192]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 小家鼠]]> <![CDATA[<400> 123]]> Val Thr Ala Arg Arg Leu Asn Cys Val Lys His Thr Tyr Pro Ser Gly 1 5 10 15 His Lys Cys Cys Arg Glu Cys Gln Pro Gly His Gly Met Val Ser Arg 20 25 30 Cys Asp His Thr Arg Asp Thr Leu Cys His Pro Cys Glu Thr Gly Phe 35 40 45 Tyr Asn Glu Ala Val Asn Tyr Asp Thr Cys Lys Gln Cys Thr Gln Cys 50 55 60 Asn His Arg Ser Gly Ser Glu Leu Lys Gln Asn Cys Thr Pro Thr Gln 65 70 75 80 Asp Thr Val Cys Arg Cys Arg Pro Gly Thr Gln Pro Arg Gln Asp Ser 85 90 95 Gly Tyr Lys Leu Gly Val Asp Cys Val Pro Cys Pro Pro Gly His Phe 100 105 110 Ser Pro Gly Asn Asn Gln Ala Cys Lys Pro Trp Thr Asn Cys Thr Leu 115 120 125 Ser Gly Lys Gln Thr Arg His Pro Ala Ser Asp Ser Leu Asp Ala Val 130 135 140 Cys Glu Asp Arg Ser Leu Leu Ala Thr Leu Leu Trp Glu Thr Gln Arg 145 150 155 160 Pro Thr Phe Arg Pro Thr Thr Val Gln Ser Thr Thr Val Trp Pro Arg 165 170 175 Thr Ser Glu Leu Pro Ser Pro Pro Thr Leu Val Thr Pro Glu Gly Pro 180 185 190 <![CDATA[<210> 124]]> <![CDATA[<211> 681]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 核苷酸序列Fc臼鏈]]> <![CDATA[<400> 124]]> gacaaaactc acacatgccc accgtgccca gcacctgaac tcctgggggg accgtcagtc 60 ttcctcttcc ccccaaaacc caaggacacc ctcatgatct cccggacccc tgaggtcaca 120 tgcgtggtgg tggacgtgag ccacgaagac cctgaggtca agttcaactg gtacgtggac 180 ggcgtggagg tgcataatgc caagacaaag ccgcgggagg agcagtacaa cagcacgtac 240 cgtgtggtca gcgtcctcac cgtcctgcac caggactggc tgaatggcaa ggagtacaag 300 tgcaaggtct ccaacaaagc cctcccagcc cccatcgaga aaaccatctc caaagccaaa 360 gggcagcccc gagaaccaca ggtgtgcacc ctgcccccat cccgggatga gctgaccaag 420 aaccaggtca gcctctcgtg cgcagtcaaa ggcttctatc ccagcgacat cgccgtggag 480 tgggagagca atgggcagcc ggagaacaac tacaagacca cgcctcccgt gctggactcc 540 gacggctcct tcttcctcgt gagcaagctc accgtggaca agagcaggtg gcagcagggg 600 aacgtcttct catgctccgt gatgcatgag gctctgcaca accactacac gcagaagagc 660 ctctccctgt ctccgggtaa a 681 <![CDATA[<210> 125]]> <![CDATA[<211> 1335]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 核苷酸序列人類OX40抗原Fc杵鏈]]> <![CDATA[<400> 125]]> ctgcactgcg tgggcgacac ctaccccagc aacgaccggt gctgccacga gtgcagaccc 60 ggcaacggca tggtgtcccg gtgcagccgg tcccagaaca ccgtgtgcag accttgcggc 120 cctggcttct acaacgacgt ggtgtccagc aagccctgca agccttgtac ctggtgcaac 180 ctgcggagcg gcagcgagcg gaagcagctg tgtaccgcca cccaggatac cgtgtgccgg 240 tgtagagccg gcacccagcc cctggacagc tacaaacccg gcgtggactg cgccccttgc 300 cctcctggcc acttcagccc tggcgacaac caggcctgca agccttggac caactgcacc 360 ctggccggca agcacaccct gcagcccgcc agcaatagca gcgacgccat ctgcgaggac 420 cgggatcctc ctgccaccca gcctcaggaa acccagggcc ctcccgccag acccatcacc 480 gtgcagccta cagaggcctg gcccagaacc agccaggggc ctagcaccag acccgtggaa 540 gtgcctggcg gcagagccgt cgacgaacag ttatattttc agggcggctc acccaaatct 600 gcagacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 660 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 720 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 780 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 840 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 900 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 960 aaagggcagc cccgagaacc acaggtgtac accctgcccc catgccggga tgagctgacc 1020 aagaaccagg tcagcctgtg gtgcctggtc aaaggcttct atcccagcga catcgccgtg 1080 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1140 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1200 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1260 agcctctccc tgtctccggg taaatccgga ggcctgaacg acatcttcga ggcccagaag 1320 attgaatggc acgag 1335 <![CDATA[<210> 126]]> <![CDATA[<211> 1335]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 核苷酸序列獼猴OX40抗原Fc杵鏈]]> <![CDATA[<400> 126]]> ctccactgtg tcggggacac ctaccccagc aacgaccggt gctgtcagga gtgcaggcca 60 ggcaacggga tggtgagccg ctgcaaccgc tcccagaaca cggtgtgccg tccgtgcggg 120 cccggcttct acaacgacgt ggtcagcgcc aagccctgca aggcctgcac atggtgcaac 180 ctcagaagtg ggagtgagcg gaaacagccg tgcacggcca cacaggacac agtctgccgc 240 tgccgggcgg gcacccagcc cctggacagc tacaagcctg gagttgactg tgccccctgc 300 cctccagggc acttctcccc gggcgacaac caggcctgca agccctggac caactgcacc 360 ttggccggga agcacaccct gcagccagcc agcaatagct cggacgccat ctgtgaggac 420 agggaccccc cacccacaca gccccaggag acccagggcc ccccggccag gcccaccact 480 gtccagccca ctgaagcctg gcccagaacc tcacagagac cctccacccg gcccgtggag 540 gtccccaggg gccctgcggt cgacgaacag ttatattttc agggcggctc acccaaatct 600 gcagacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 660 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 720 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 780 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 840 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 900 aagtgcaagg tctccaacaa agccctccca gcccccatcg agaaaaccat ctccaaagcc 960 aaagggcagc cccgagaacc acaggtgtac accctgcccc catgccggga tgagctgacc 1020 aagaaccagg tcagcctgtg gtgcctggtc aaaggcttct atcccagcga catcgccgtg 1080 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1140 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1200 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1260 agcctctccc tgtctccggg taaatccgga ggcctgaacg acatcttcga ggcccagaag 1320 attgaatggc acgag 1335 <![CDATA[<210> 127]]> <![CDATA[<211> 1353]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 核苷酸序列鼠類OX40抗原Fc杵鏈]]> <![CDATA[<400> 127]]> gtgaccgcca gacggctgaa ctgcgtgaag cacacctacc ccagcggcca caagtgctgc 60 agagagtgcc agcccggcca cggcatggtg tccagatgcg accacacacg ggacaccctg 120 tgccaccctt gcgagacagg cttctacaac gaggccgtga actacgatac ctgcaagcag 180 tgcacccagt gcaaccacag aagcggcagc gagctgaagc agaactgcac ccccacccag 240 gataccgtgt gcagatgcag acccggcacc cagcccagac aggacagcgg ctacaagctg 300 ggcgtggact gcgtgccctg ccctcctggc cacttcagcc ccggcaacaa ccaggcctgc 360 aagccctgga ccaactgcac cctgagcggc aagcagacca gacaccccgc cagcgacagc 420 ctggatgccg tgtgcgagga cagaagcctg ctggccaccc tgctgtggga gacacagcgg 480 cccaccttca gacccaccac cgtgcagagc accaccgtgt ggcccagaac cagcgagctg 540 cccagtcctc ctaccctcgt gacacctgag ggccccgtcg acgaacagtt atattttcag 600 ggcggctcac ccaaatctgc agacaaaact cacacatgcc caccgtgccc agcacctgaa 660 ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac cctcatgatc 720 tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga ccctgaggtc 780 aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa gccgcgggag 840 gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca ccaggactgg 900 ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc ccccatcgag 960 aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtacac cctgccccca 1020 tgccgggatg agctgaccaa gaaccaggtc agcctgtggt gcctggtcaa aggcttctat 1080 cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa ctacaagacc 1140 acgcctcccg tgctggactc cgacggctcc ttcttcctct acagcaagct caccgtggac 1200 aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga ggctctgcac 1260 aaccactaca cgcagaagag cctctccctg tctccgggta aatccggagg cctgaacgac 1320 atcttcgagg cccagaagat tgaatggcac gag 1353 <![CDATA[<210> 128]]> <![CDATA[<211> 227]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fc臼鏈]]> <![CDATA[<400> 128]]> Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly 1 5 10 15 Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met 20 25 30 Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His 35 40 45 Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val 50 55 60 His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr 65 70 75 80 Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly 85 90 95 Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile 100 105 110 Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val 115 120 125 Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser 130 135 140 Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu 145 150 155 160 Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro 165 170 175 Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val 180 185 190 Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met 195 200 205 His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser 210 215 220 Pro Gly Lys 225 <![CDATA[<210> 129]]> <![CDATA[<211> 445]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 人類OX40抗原Fc杵鏈]]> <![CDATA[<400> 129]]> Leu His Cys Val Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys His 1 5 10 15 Glu Cys Arg Pro Gly Asn Gly Met Val Ser Arg Cys Ser Arg Ser Gln 20 25 30 Asn Thr Val Cys Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val 35 40 45 Ser Ser Lys Pro Cys Lys Pro Cys Thr Trp Cys Asn Leu Arg Ser Gly 50 55 60 Ser Glu Arg Lys Gln Leu Cys Thr Ala Thr Gln Asp Thr Val Cys Arg 65 70 75 80 Cys Arg Ala Gly Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp 85 90 95 Cys Ala Pro Cys Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala 100 105 110 Cys Lys Pro Trp Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln 115 120 125 Pro Ala Ser Asn Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro 130 135 140 Ala Thr Gln Pro Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Ile Thr 145 150 155 160 Val Gln Pro Thr Glu Ala Trp Pro Arg Thr Ser Gln Gly Pro Ser Thr 165 170 175 Arg Pro Val Glu Val Pro Gly Gly Arg Ala Val Asp Glu Gln Leu Tyr 180 185 190 Phe Gln Gly Gly Ser Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro 195 200 205 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 210 215 220 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 225 230 235 240 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 245 250 255 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 260 265 270 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 275 280 285 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 290 295 300 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 305 310 315 320 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg 325 330 335 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly 340 345 350 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 355 360 365 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 370 375 380 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 385 390 395 400 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 405 410 415 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Ser Gly Gly Leu 420 425 430 Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 435 440 445 <![CDATA[<210> 130]]> <![CDATA[<211> 445]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 獼猴OX40抗原]]> Fc杵鏈 <![CDATA[<400> 130]]> Leu His Cys Val Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys Gln 1 5 10 15 Glu Cys Arg Pro Gly Asn Gly Met Val Ser Arg Cys Asn Arg Ser Gln 20 25 30 Asn Thr Val Cys Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val 35 40 45 Ser Ala Lys Pro Cys Lys Ala Cys Thr Trp Cys Asn Leu Arg Ser Gly 50 55 60 Ser Glu Arg Lys Gln Pro Cys Thr Ala Thr Gln Asp Thr Val Cys Arg 65 70 75 80 Cys Arg Ala Gly Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp 85 90 95 Cys Ala Pro Cys Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala 100 105 110 Cys Lys Pro Trp Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln 115 120 125 Pro Ala Ser Asn Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro 130 135 140 Pro Thr Gln Pro Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Thr Thr 145 150 155 160 Val Gln Pro Thr Glu Ala Trp Pro Arg Thr Ser Gln Arg Pro Ser Thr 165 170 175 Arg Pro Val Glu Val Pro Arg Gly Pro Ala Val Asp Glu Gln Leu Tyr 180 185 190 Phe Gln Gly Gly Ser Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro 195 200 205 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 210 215 220 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 225 230 235 240 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 245 250 255 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 260 265 270 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 275 280 285 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 290 295 300 Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 305 310 315 320 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg 325 330 335 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly 340 345 350 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 355 360 365 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 370 375 380 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 385 390 395 400 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 405 410 415 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Ser Gly Gly Leu 420 425 430 Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp His Glu 435 440 445 <![CDATA[<210> 131]]> <![CDATA[<211> 451]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類OX40抗原Fc杵鏈]]> <![CDATA[<400> 131]]> Val Thr Ala Arg Arg Leu Asn Cys Val Lys His Thr Tyr Pro Ser Gly 1 5 10 15 His Lys Cys Cys Arg Glu Cys Gln Pro Gly His Gly Met Val Ser Arg 20 25 30 Cys Asp His Thr Arg Asp Thr Leu Cys His Pro Cys Glu Thr Gly Phe 35 40 45 Tyr Asn Glu Ala Val Asn Tyr Asp Thr Cys Lys Gln Cys Thr Gln Cys 50 55 60 Asn His Arg Ser Gly Ser Glu Leu Lys Gln Asn Cys Thr Pro Thr Gln 65 70 75 80 Asp Thr Val Cys Arg Cys Arg Pro Gly Thr Gln Pro Arg Gln Asp Ser 85 90 95 Gly Tyr Lys Leu Gly Val Asp Cys Val Pro Cys Pro Pro Gly His Phe 100 105 110 Ser Pro Gly Asn Asn Gln Ala Cys Lys Pro Trp Thr Asn Cys Thr Leu 115 120 125 Ser Gly Lys Gln Thr Arg His Pro Ala Ser Asp Ser Leu Asp Ala Val 130 135 140 Cys Glu Asp Arg Ser Leu Leu Ala Thr Leu Leu Trp Glu Thr Gln Arg 145 150 155 160 Pro Thr Phe Arg Pro Thr Thr Val Gln Ser Thr Thr Val Trp Pro Arg 165 170 175 Thr Ser Glu Leu Pro Ser Pro Pro Thr Leu Val Thr Pro Glu Gly Pro 180 185 190 Val Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Pro Lys Ser Ala Asp 195 200 205 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 210 215 220 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 225 230 235 240 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 245 250 255 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 260 265 270 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 275 280 285 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 290 295 300 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 305 310 315 320 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 325 330 335 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 340 345 350 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 355 360 365 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 370 375 380 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 385 390 395 400 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 405 410 415 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 420 425 430 Gly Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu 435 440 445 Trp His Glu 450 <![CDATA[<210> 132]]> <![CDATA[<211> 1613]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 文庫DP88-4之核苷酸序列]]> <![CDATA[<400> 132]]> tgaaatacct attgcctacg gcagccgctg gattgttatt actcgcggcc cagccggcca 60 tggccgacat ccagatgacc cagtctcctt ccaccctgtc tgcatctgta ggagaccgtg 120 tcaccatcac ttgccgtgcc agtcagagta ttagtagctg gttggcctgg tatcagcaga 180 aaccagggaa agcccctaag ctcctgatct atgatgcctc cagtttggaa agtggggtcc 240 catcacgttt cagcggcagt ggatccggga cagaattcac tctcaccatc agcagcttgc 300 agcctgatga ttttgcaact tattactgcc aacagtataa tagttattct acgtttggcc 360 agggcaccaa agtcgagatc aagcgtacgg tggctgcacc atctgtcttc atcttcccgc 420 catctgatga gcagttgaaa tctggaactg cctctgttgt gtgcctgctg aataacttct 480 atcccagaga ggccaaagta cagtggaagg tggataacgc cctccaatcg ggtaactccc 540 aggagagtgt cacagagcag gacagcaagg acagcaccta cagcctcagc agcaccctga 600 cgctgagcaa agcagactac gagaaacaca aagtctacgc ctgcgaagtc acccatcagg 660 gcctgagctc gcccgtcaca aagagcttca acaggggaga gtgtggagcc gcagaacaaa 720 aactcatctc agaagaggat ctgaatggag ccgcagacta caaggacgac gacgacaagg 780 gtgccgcata ataaggcgcg ccaattctat ttcaaggaga cagtcatatg aaatacctgc 840 tgccgaccgc tgctgctggt ctgctgctcc tcgctgccca gccggcgatg gcccaggtgc 900 aattggtgca gtctggggct gaggtgaaga agcctgggtc ctcggtgaag gtctcctgca 960 aggcctccgg aggcacattc agcagctacg ctataagctg ggtgcgacag gcccctggac 1020 aagggctcga gtggatggga gggatcatcc ctatctttgg tacagcaaac tacgcacaga 1080 agttccaggg cagggtcacc attactgcag acaaatccac gagcacagcc tacatggagc 1140 tgagcagcct gagatctgag gacaccgccg tgtattactg tgcgagacta tccccaggcg 1200 gttactatgt tatggatgcc tggggccaag ggaccaccgt gaccgtctcc tcagctagca 1260 ccaaaggccc atcggtcttc cccctggcac cctcctccaa gagcacctct gggggcacag 1320 cggccctggg ctgcctggtc aaggactact tccccgaacc ggtgacggtg tcgtggaact 1380 caggcgccct gaccagcggc gtgcacacct tcccggctgt cctacagtcc tcaggactct 1440 actccctcag cagcgtggtg accgtgccct ccagcagctt gggcacccag acctacatct 1500 gcaacgtgaa tcacaagccc agcaacacca aagtggacaa gaaagttgag cccaaatctt 1560 gtgacgcggc cgcaagcact agtgcccatc accatcacca tcacgccgcg gca 1613 <![CDATA[<210> 133]]> <![CDATA[<211> 723]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab輕鏈Vk1_5之核苷酸序列]]> <![CDATA[<400> 133]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tataatagtt attctacgtt tggccagggc 300 accaaagtcg agatcaagcg tacggtggct gcaccatctg tcttcatctt cccgccatct 360 gatgagcagt tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc 420 agagaggcca aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag 480 agtgtcacag agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg 540 agcaaagcag actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg 600 agctcgcccg tcacaaagag cttcaacagg ggagagtgtg gagccgcaga acaaaaactc 660 atctcagaag aggatctgaa tggagccgca gactacaagg acgacgacga caagggtgcc 720 gca 723 <![CDATA[<210> 134]]> <![CDATA[<211> 241]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab輕鏈Vk1_5]]> <![CDATA[<400> 134]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Ser Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys Gly Ala Ala Glu Gln Lys Leu Ile Ser Glu Glu 210 215 220 Asp Leu Asn Gly Ala Ala Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala 225 230 235 240 Ala <![CDATA[<210> 135]]> <![CDATA[<211> 720]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab重鏈VH1_69之核苷酸序列]]> <![CDATA[<400> 135]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagactatcc 300 ccaggcggtt actatgttat ggatgcctgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca aaggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaaag tggacaagaa agttgagccc 660 aaatcttgtg acgcggccgc aagcactagt gcccatcacc atcaccatca cgccgcggca 720 <![CDATA[<210> 136]]> <![CDATA[<211> 240]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab重鏈VH1_69]]> <![CDATA[<400> 136]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Leu Ser Pro Gly Gly Tyr Tyr Val Met Asp Ala Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Ala Ala Ala Ser Thr Ser Ala His His His His His His Ala Ala Ala 225 230 235 240 <![CDATA[<210> 137]]> <![CDATA[<211> 26]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> LMB3引子]]> <![CDATA[<400> 137]]> caggaaacag ctatgaccat gattac 26 <![CDATA[<210> 138]]> <![CDATA[<211> 71]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vk1_5_L3r_S引子]]> <![CDATA[<400> 138]]> ctcgactttg gtgccctggc caaacgtsba atacgaatta tactgttggc agtaataagt 60 tgcaaaatca t 71 <![CDATA[<210> 139]]> <![CDATA[<211> 74]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vk1_5_L3r_SY引子]]> <![CDATA[<400> 139]]> ctcgactttg gtgccctggc caaacgtmhr sgratacgaa ttatactgtt ggcagtaata 60 agttgcaaaa tcat 74 <![CDATA[<210> 140]]> <![CDATA[<211> 77]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vk1_5_L3r_SPY]]> <![CDATA[<400> 140]]> ctcgactttg gtgccctggc caaacgtmhh msssgratac gaattatact gttggcagta 60 ataagttgca aaatcat 77 <![CDATA[<210> 141]]> <![CDATA[<211> 27]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> RJH31引子]]> <![CDATA[<400> 141]]> acgtttggcc agggcaccaa agtcgag 27 <![CDATA[<210> 142]]> <![CDATA[<211> 28]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> RJH32引子]]> <![CDATA[<400> 142]]> tctcgcacag taatacacgg cggtgtcc 28 <![CDATA[<210> 143]]> <![CDATA[<211> 64]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP88-v4-4]]> <![CDATA[<400> 143]]> ggacaccgcc gtgtattact gtgcgagaga ctactggggc caagggacca ccgtgaccgt 60 ctcc 64 <![CDATA[<210> 144]]> <![CDATA[<211> 64]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP88-v4-6]]> <![CDATA[<400> 144]]> ggacaccgcc gtgtattact gtgcgagaga ctactggggc caagggacca ccgtgaccgt 60 ctcc 64 <![CDATA[<210> 145]]> <![CDATA[<211> 64]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP88-v4-8]]> <![CDATA[<400> 145]]> ggacaccgcc gtgtattact gtgcgagaga ctactggggc caagggacca ccgtgaccgt 60 ctcc 64 <![CDATA[<210> 146]]> <![CDATA[<211> 30]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> fdseqlong引子]]> <![CDATA[<400> 146]]> gacgttagta aatgaatttt ctgtatgagg 30 <![CDATA[<210> 147]]> <![CDATA[<211> 1596]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (Vk3_20/VH3_23)模板]]> <![CDATA[<400> 147]]> atgaaatacc tattgcctac ggcagccgct ggattgttat tactcgcggc ccagccggcc 60 atggccgaaa tcgtgttaac gcagtctcca ggcaccctgt ctttgtctcc aggggaaaga 120 gccaccctct cttgcagggc cagtcagagt gttagcagca gctacttagc ctggtaccag 180 cagaaacctg gccaggctcc caggctcctc atctatggag catccagcag ggccactggc 240 atcccagaca ggttcagtgg cagtggatcc gggacagact tcactctcac catcagcaga 300 ctggagcctg aagattttgc agtgtattac tgtcagcagt atggtagctc accgctgacg 360 ttcggccagg ggaccaaagt ggaaatcaaa cgtacggtgg ctgcaccatc tgtcttcatc 420 ttcccgccat ctgatgagca gttgaaatct ggaactgcct ctgttgtgtg cctgctgaat 480 aacttctatc ccagagaggc caaagtacag tggaaggtgg ataacgccct ccaatcgggt 540 aactcccagg agagtgtcac agagcaggac agcaaggaca gcacctacag cctcagcagc 600 accctgacgc tgagcaaagc agactacgag aaacacaaag tctacgcctg cgaagtcacc 660 catcagggcc tgagctcgcc cgtcacaaag agcttcaaca ggggagagtg tggagccgca 720 catcaccatc accatcacgg agccgcagac tacaaggacg acgacgacaa gggtgccgca 780 taataaggcg cgccaattct atttcaagga gacagtcata tgaaatacct gctgccgacc 840 gctgctgctg gtctgctgct cctcgctgcc cagccggcga tggccgaggt gcaattgctg 900 gagtctgggg gaggcttggt acagcctggg gggtccctga gactctcctg tgcagcctcc 960 ggattcacct ttagcagtta tgccatgagc tgggtccgcc aggctccagg gaaggggctg 1020 gagtgggtct cagctattag tggtagtggt ggtagcacat actacgcaga ctccgtgaag 1080 ggccggttca ccatctccag agacaattcc aagaacacgc tgtatctgca gatgaacagc 1140 ctgagagccg aggacacggc cgtatattac tgtgcgaaac cgtttccgta ttttgactac 1200 tggggccaag gaaccctggt caccgtctcg agtgctagca ccaaaggccc atcggtcttc 1260 cccctggcac cctcctccaa gagcacctct gggggcacag cggccctggg ctgcctggtc 1320 aaggactact tccccgaacc ggtgacggtg tcgtggaact caggcgccct gaccagcggc 1380 gtgcacacct tcccggctgt cctacagtcc tcaggactct actccctcag cagcgtggtg 1440 accgtgccct ccagcagctt gggcacccag acctacatct gcaacgtgaa tcacaagccc 1500 agcaacacca aagtggacaa gaaagttgag cccaaatctt gtgacgcggc cgcagaacaa 1560 aaactcatct cagaagagga tctgaatgcc gcggca 1596 <![CDATA[<210> 148]]> <![CDATA[<211> 714]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab輕鏈Vk3_20]]> <![CDATA[<400> 148]]> gaaatcgtgt taacgcagtc tccaggcacc ctgtctttgt ctccagggga aagagccacc 60 ctctcttgca gggccagtca gagtgttagc agcagctact tagcctggta ccagcagaaa 120 cctggccagg ctcccaggct cctcatctat ggagcatcca gcagggccac tggcatccca 180 gacaggttca gtggcagtgg atccgggaca gacttcactc tcaccatcag cagactggag 240 cctgaagatt ttgcagtgta ttactgtcag cagtatggta gctcaccgct gacgttcggc 300 caggggacca aagtggaaat caaacgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgtggagc cgcacatcac 660 catcaccatc acggagccgc agactacaag gacgacgacg acaagggtgc cgca 714 <![CDATA[<210> 149]]> <![CDATA[<211> 238]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab輕鏈Vk3_20]]> <![CDATA[<400> 149]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys Gly Ala Ala His His His His His His 210 215 220 Gly Ala Ala Asp Tyr Lys Asp Asp Asp Asp Lys Gly Ala Ala 225 230 235 <![CDATA[<210> 150]]> <![CDATA[<211> 711]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab重鏈VH3_23]]> <![CDATA[<400> 150]]> gaggtgcaat tgctggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gaaaccgttt 300 ccgtattttg actactgggg ccaaggaacc ctggtcaccg tctcgagtgc tagcaccaaa 360 ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420 ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480 gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540 ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600 gtgaatcaca agcccagcaa caccaaagtg gacaagaaag ttgagcccaa atcttgtgac 660 gcggccgcag aacaaaaact catctcagaa gaggatctga atgccgcggc a 711 <![CDATA[<210> 151]]> <![CDATA[<211> 237]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab重鏈VH3_23 (DP47)]]> <![CDATA[<400> 151]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Lys Pro Phe Pro Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Ala Ala Ala Glu 210 215 220 Gln Lys Leu Ile Ser Glu Glu Asp Leu Asn Ala Ala Ala 225 230 235 <![CDATA[<210> 152]]> <![CDATA[<211> 25]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> MS64引子]]> <![CDATA[<400> 152]]> acgttcggcc aggggaccaa agtgg 25 <![CDATA[<210> 153]]> <![CDATA[<211> 25]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP47CDR3_ba引子]]> <![CDATA[<400> 153]]> cgcacagtaa tatacggccg tgtcc 25 <![CDATA[<210> 154]]> <![CDATA[<211> 64]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP47-v4-4]]> <![CDATA[<400> 154]]> cgaggacacg gccgtatatt actgtgcgga ctactggggc caaggaaccc tggtcaccgt 60 ctcg 64 <![CDATA[<210> 155]]> <![CDATA[<211> 64]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP47-v4-6]]> <![CDATA[<400> 155]]> cgaggacacg gccgtatatt actgtgcgga ctactggggc caaggaaccc tggtcaccgt 60 ctcg 64 <![CDATA[<210> 156]]> <![CDATA[<211> 64]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DP47-v4-8]]> <![CDATA[<400> 156]]> cgaggacacg gccgtatatt actgtgcgga ctactggggc caaggaaccc tggtcaccgt 60 ctcg 64 <![CDATA[<210> 157]]> <![CDATA[<211> 30]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> fdseqlong]]> <![CDATA[<400> 157]]> gacgttagta aatgaatttt ctgtatgagg 30 <![CDATA[<210> 158]]> <![CDATA[<211> 1605]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vl3_19/VH3_23文庫模板]]> <![CDATA[<400> 158]]> atgaaatacc tattgcctac ggcagccgct ggattgttat tactcgcggc ccagccggcc 60 atggcctcgt ctgagctgac tcaggaccct gctgtgtctg tggccttggg acagacagtc 120 aggatcacat gccaaggaga cagcctcaga agttattatg caagctggta ccagcagaag 180 ccaggacagg cccctgtact tgtcatctat ggtaaaaaca accggccctc agggatccca 240 gaccgattct ctggctccag ctcaggaaac acagcttcct tgaccatcac tggggctcag 300 gcggaagatg aggctgacta ttactgtaac tcccgtgata gtagcggtaa tcatgtggta 360 ttcggcggag ggaccaagct gaccgtccta ggacaaccca aggctgcccc cagcgtgacc 420 ctgttccccc ccagcagcga ggaattgcag gccaacaagg ccaccctggt ctgcctgatc 480 agcgacttct acccaggcgc cgtgaccgtg gcctggaagg ccgacagcag ccccgtgaag 540 gccggcgtgg agaccaccac ccccagcaag cagagcaaca acaagtacgc cgccagcagc 600 tacctgagcc tgacccccga gcagtggaag agccacaggt cctacagctg ccaggtgacc 660 cacgagggca gcaccgtgga gaaaaccgtg gcccccaccg agtgcagcgg agccgcagaa 720 caaaaactca tctcagaaga ggatctgaat ggagccgcag actacaagga cgacgacgac 780 aagggtgccg cataataagg cgcgccaatt ctatttcaag gagacagtca tatgaaatac 840 ctgctgccga ccgctgctgc tggtctgctg ctcctcgctg cccagccggc gatggccgag 900 gtgcaattgc tggagtctgg gggaggcttg gtacagcctg gggggtccct gagactctcc 960 tgtgcagcct ccggattcac ctttagcagt tatgccatga gctgggtccg ccaggctcca 1020 gggaaggggc tggagtgggt ctcagctatt agtggtagtg gtggtagcac atactacgca 1080 gactccgtga agggccggtt caccatctcc agagacaatt ccaagaacac gctgtatctg 1140 cagatgaaca gcctgagagc cgaggacacg gccgtatatt actgtgcgaa accgtttccg 1200 tattttgact actggggcca aggaaccctg gtcaccgtct cgagtgctag caccaaaggc 1260 ccatcggtct tccccctggc accctcctcc aagagcacct ctgggggcac agcggccctg 1320 ggctgcctgg tcaaggacta cttccccgaa ccggtgacgg tgtcgtggaa ctcaggcgcc 1380 ctgaccagcg gcgtgcacac cttcccggct gtcctacagt cctcaggact ctactccctc 1440 agcagcgtgg tgaccgtgcc ctccagcagc ttgggcaccc agacctacat ctgcaacgtg 1500 aatcacaagc ccagcaacac caaagtggac aagaaagttg agcccaaatc ttgtgacgcg 1560 gccgcaagca ctagtgccca tcaccatcac catcacgccg cggca 1605 <![CDATA[<210> 159]]> <![CDATA[<211> 726]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab輕鏈Vl3_19]]> <![CDATA[<400> 159]]> tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60 acatgccaag gagacagcct cagaagttat tatgcaagct ggtaccagca gaagccagga 120 caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga 180 ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa 240 gatgaggctg actattactg taactcccgt gatagtagcg gtaatcatgt ggtattcggc 300 ggagggacca agctgaccgt cctaggacaa cccaaggctg cccccagcgt gaccctgttc 360 ccccccagca gcgaggaatt gcaggccaac aaggccaccc tggtctgcct gatcagcgac 420 ttctacccag gcgccgtgac cgtggcctgg aaggccgaca gcagccccgt gaaggccggc 480 gtggagacca ccacccccag caagcagagc aacaacaagt acgccgccag cagctacctg 540 agcctgaccc ccgagcagtg gaagagccac aggtcctaca gctgccaggt gacccacgag 600 ggcagcaccg tggagaaaac cgtggccccc accgagtgca gcggagccgc agaacaaaaa 660 ctcatctcag aagaggatct gaatggagcc gcagactaca aggacgacga cgacaagggt 720 gccgca 726 <![CDATA[<210> 160]]> <![CDATA[<211> 242]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Fab輕鏈Vl3_19]]> <![CDATA[<400> 160]]> Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Asp Ser Ser Gly Asn His 85 90 95 Val Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys 100 105 110 Ala Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln 115 120 125 Ala Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly 130 135 140 Ala Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly 145 150 155 160 Val Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala 165 170 175 Ser Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser 180 185 190 Tyr Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val 195 200 205 Ala Pro Thr Glu Cys Ser Gly Ala Ala Glu Gln Lys Leu Ile Ser Glu 210 215 220 Glu Asp Leu Asn Gly Ala Ala Asp Tyr Lys Asp Asp Asp Asp Lys Gly 225 230 235 240 Ala Ala <![CDATA[<210> 161]]> <![CDATA[<211> 26]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> LMB3 λ-DP47文庫]]> <![CDATA[<400> 161]]> caggaaacag ctatgaccat gattac 26 <![CDATA[<210> 162]]> <![CDATA[<211> 85]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vl_3_19_L3r_V λ-DP47文庫]]> <![CDATA[<400> 162]]> ggacggtcag cttggtccct ccgccgaata cvhvattacc gctactatca cgggagttac 60 agtaatagtc agcctcatct tccgc 85 <![CDATA[<210> 163]]> <![CDATA[<211> 88]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vl_3_19_L3r_HV λ-DP47]]> <![CDATA[<400> 163]]> ggacggtcag cttggtccct ccgccgaata ccmmatgatt accgctacta tcacgggagt 60 tacagtaata gtcagcctca tcttccgc 88 <![CDATA[<210> 164]]> <![CDATA[<211> 91]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> Vl_3_19_L3r_HLV λ-DP47]]> <![CDATA[<400> 164]]> ggacggtcag cttggtccct ccgccgaata crhmvwgatg attaccgcta ctatcacggg 60 agttacagta atagtcagcc tcatcttccg c 91 <![CDATA[<210> 165]]> <![CDATA[<211> 28]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> RJH80 λ-DP47]]> <![CDATA[<400> 165]]> ttcggcggag ggaccaagct gaccgtcc 28 <![CDATA[<210> 166]]> <![CDATA[<211> 318]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (8H9 ) VL]]> <![CDATA[<400> 166]]> tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60 acatgccaag gagacagcct cagaagttat tatgcaagct ggtaccagca gaagccagga 120 caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga 180 ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa 240 gatgaggctg actattactg taactcccgt gttatgcctc ataatcgcgt attcggcgga 300 gggaccaagc tgaccgtc 318 <![CDATA[<210> 167]]> <![CDATA[<211> 360]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (8H9) VH]]> <![CDATA[<400> 167]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgttttc 300 taccgtggtg gtgtttctat ggactactgg ggccaaggaa ccctggtcac cgtctcgagt 360 <![CDATA[<210> 168]]> <![CDATA[<211> 321]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (49B4) VL]]> <![CDATA[<400> 168]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatagttcgc agccgtatac gtttggccag 300 ggcaccaaag tcgagatcaa g 321 <![CDATA[<210> 169]]> <![CDATA[<211> 354]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (49B4) VH]]> <![CDATA[<400> 169]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctca 354 <![CDATA[<210> 170]]> <![CDATA[<211> 324]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (1G4) VL]]> <![CDATA[<400> 170]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatatttcgt attccatgtt gacgtttggc 300 cagggcacca aagtcgagat caag 324 <![CDATA[<210> 171]]> <![CDATA[<211> 348]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (1G4) VH]]> <![CDATA[<400> 171]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttctatgg actactgggg ccaagggacc accgtgaccg tctcctca 348 <![CDATA[<210> 172]]> <![CDATA[<211> 321]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (20B7) VL]]> <![CDATA[<400> 172]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatcaggctt tttcgcttac gtttggccag 300 ggcaccaaag tcgagatcaa g 321 <![CDATA[<210> 173]]> <![CDATA[<211> 366]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (20B7) VH]]> <![CDATA[<400> 173]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagttaac 300 tacccgtact cttactgggg tgacttcgac tactggggcc aagggaccac cgtgaccgtc 360 tcctca 366 <![CDATA[<210> 174]]> <![CDATA[<211> 324]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-563) VL]]> <![CDATA[<400> 174]]> gagatcgtgc tgacccagag ccccggcaca ctctccctgt ctcctgggga aagggccacc 60 ctttcatgca gagccagcca gtccgtctct agtagctacc tggcatggta tcagcagaag 120 ccaggacaag ccccccgcct cctgatttac ggcgcttcct ctcgggcaac tggtatccct 180 gacaggttct cagggagcgg aagcggaaca gattttacct tgactatttc tagactggag 240 ccagaggact tcgccgtgta ttactgtcag cagtacggta gtagccccct cacctttggc 300 caggggacaa aagtcgaaat caag 324 <![CDATA[<210> 175]]> <![CDATA[<211> 348]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-563) VH]]> <![CDATA[<400> 175]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcttgacgtt 300 ggtgctttcg actactgggg ccaaggagcc ctggtcaccg tctcgagt 348 <![CDATA[<210> 176]]> <![CDATA[<211> 324]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-564) VL]]> <![CDATA[<400> 176]]> gagatcgtgc tgacccagag ccccggcaca ctctccctgt ctcctgggga aagggccacc 60 ctttcatgca gagccagcca gtccgtctct agtagctacc tggcatggta tcagcagaag 120 ccaggacaag ccccccgcct cctgatttac ggcgcttcct ctcgggcaac tggtatccct 180 gacaggttct cagggagcgg aagcggaaca gattttacct tgactatttc tagactggag 240 ccagaggact tcgccgtgta ttactgtcag cagtacggta gtagccccct cacctttggc 300 caggggacaa aagtcgaaat caag 324 <![CDATA[<210> 177]]> <![CDATA[<211> 348]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-564) VH]]> <![CDATA[<400> 177]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gttcgacgtt 300 ggtccgttcg actactgggg ccaaggaacc ctggtcaccg tctcgagt 348 <![CDATA[<210> 178]]> <![CDATA[<211> 318]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (17A9) VL]]> <![CDATA[<400> 178]]> tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60 acatgccaag gagacagcct cagaagttat tatgcaagct ggtaccagca gaagccagga 120 caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga 180 ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa 240 gatgaggctg actattactg taactcccgt gttatgcctc ataatcgcgt attcggcgga 300 gggaccaagc tgaccgtc 318 <![CDATA[<210> 179]]> <![CDATA[<211> 360]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (17A9) VH]]> <![CDATA[<400> 179]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgttttc 300 taccgtggtg gtgtttctat ggactactgg ggccaaggaa ccctggtcac cgtctcgagt 360 <![CDATA[<210> 180]]> <![CDATA[<211> 645]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 8H9 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 180]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatttgacgt attcgcggtt tacgtttggc 300 cagggcacca aagtcgagat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <![CDATA[<210> 181]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 8H9 P329GLALA IgG1重鏈]]> <![CDATA[<400> 181]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttggatgg actactgggg ccaagggacc accgtgaccg tctcctcagc tagcaccaag 360 ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420 ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480 gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540 ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600 gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660 aaaactcaca catgcccacc gtgcccagca cctgaagctg cagggggacc gtcagtcttc 720 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatccc gggatgagct gaccaagaac 1080 caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 182]]> <![CDATA[<211> 215]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 8H9 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 182]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Leu Thr Tyr Ser Arg 85 90 95 Phe Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <![CDATA[<210> 183]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 8H9 P329GLALA IgG1重鏈]]> <![CDATA[<400> 183]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Trp Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 184]]> <![CDATA[<211> 642]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 49B4 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 184]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatagttcgc agccgtatac gtttggccag 300 ggcaccaaag tcgagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <![CDATA[<210> 185]]> <![CDATA[<211> 1344]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 49B4 P329GLALA IgG1重鏈]]> <![CDATA[<400> 185]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <![CDATA[<210> 186]]> <![CDATA[<211> 214]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 49B4 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 186]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ser Ser Gln Pro Tyr 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <![CDATA[<210> 187]]> <![CDATA[<211> 448]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 49B4重鏈]]> <![CDATA[<400> 187]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 188]]> <![CDATA[<211> 645]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 1G4 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 188]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatatttcgt attccatgtt gacgtttggc 300 cagggcacca aagtcgagat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <![CDATA[<210> 189]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 1G4 P329GLALA IgG1重鏈]]> <![CDATA[<400> 189]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttctatgg actactgggg ccaagggacc accgtgaccg tctcctcagc tagcaccaag 360 ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420 ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480 gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540 ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600 gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660 aaaactcaca catgcccacc gtgcccagca cctgaagctg cagggggacc gtcagtcttc 720 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatccc gggatgagct gaccaagaac 1080 caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 190]]> <![CDATA[<211> 215]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 1G4 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 190]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Ile Ser Tyr Ser Met 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <![CDATA[<210> 191]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 1G4 P329GLALA IgG1重鏈]]> <![CDATA[<400> 191]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Ser Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 192]]> <![CDATA[<211> 642]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 20B7 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 192]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatcaggctt tttcgcttac gtttggccag 300 ggcaccaaag tcgagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <![CDATA[<210> 193]]> <![CDATA[<211> 1356]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 20B7 P329GLALA IgG1重鏈]]> <![CDATA[<400> 193]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagttaac 300 tacccgtact cttactgggg tgacttcgac tactggggcc aagggaccac cgtgaccgtc 360 tcctcagcta gcaccaaggg cccatcggtc ttccccctgg caccctcctc caagagcacc 420 tctgggggca cagcggccct gggctgcctg gtcaaggact acttccccga accggtgacg 480 gtgtcgtgga actcaggcgc cctgaccagc ggcgtgcaca ccttcccggc tgtcctacag 540 tcctcaggac tctactccct cagcagcgtg gtgaccgtgc cctccagcag cttgggcacc 600 cagacctaca tctgcaacgt gaatcacaag cccagcaaca ccaaggtgga caagaaagtt 660 gagcccaaat cttgtgacaa aactcacaca tgcccaccgt gcccagcacc tgaagctgca 720 gggggaccgt cagtcttcct cttcccccca aaacccaagg acaccctcat gatctcccgg 780 acccctgagg tcacatgcgt ggtggtggac gtgagccacg aagaccctga ggtcaagttc 840 aactggtacg tggacggcgt ggaggtgcat aatgccaaga caaagccgcg ggaggagcag 900 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 960 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcg gcgcccccat cgagaaaacc 1020 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatcccgg 1080 gatgagctga ccaagaacca ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc 1140 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 1200 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1260 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1320 tacacgcaga agagcctctc cctgtctccg ggtaaa 1356 <![CDATA[<210> 194]]> <![CDATA[<211> 214]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> 20B7 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 194]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Gln Ala Phe Ser Leu 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <![CDATA[<210> 195]]> <![CDATA[<211> 452]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> 20B7 P329GLALA IgG1重鏈]]> <![CDATA[<400> 195]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Asn Tyr Pro Tyr Ser Tyr Trp Gly Asp Phe Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <![CDATA[<210> 196]]> <![CDATA[<211> 645]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> DNA CLC-563 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 196]]> gagatcgtgc tgacccagag ccccggcaca ctctccctgt ctcctgggga aagggccacc 60 ctttcatgca gagccagcca gtccgtctct agtagctacc tggcatggta tcagcagaag 120 ccaggacaag ccccccgcct cctgatttac ggcgcttcct ctcgggcaac tggtatccct 180 gacaggttct cagggagcgg aagcggaaca gattttacct tgactatttc tagactggag 240 ccagaggact tcgccgtgta ttactgtcag cagtacggta gtagccccct cacctttggc 300 caggggacaa aagtcgaaat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <![CDATA[<210> 197]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> DNA CLC-563 P329GLALA IgG1重鏈]]> <![CDATA[<400> 197]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcttgacgtt 300 ggtgctttcg actactgggg ccaaggagcc ctggtcaccg tctcgagtgc tagcaccaag 360 ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420 ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480 gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540 ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600 gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660 aaaactcaca catgcccacc gtgcccagca cctgaagctg cagggggacc gtcagtcttc 720 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatccc gggatgagct gaccaagaac 1080 caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 198]]> <![CDATA[<211> 215]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> CLC-563 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 198]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <![CDATA[<210> 199]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> CLC-563 P329GLALA IgG1重鏈]]> <![CDATA[<400> 199]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Leu Asp Val Gly Ala Phe Asp Tyr Trp Gly Gln Gly Ala Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 200]]> <![CDATA[<211> 645]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> DNA CLC-564 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 200]]> gagatcgtgc tgacccagag ccccggcaca ctctccctgt ctcctgggga aagggccacc 60 ctttcatgca gagccagcca gtccgtctct agtagctacc tggcatggta tcagcagaag 120 ccaggacaag ccccccgcct cctgatttac ggcgcttcct ctcgggcaac tggtatccct 180 gacaggttct cagggagcgg aagcggaaca gattttacct tgactatttc tagactggag 240 ccagaggact tcgccgtgta ttactgtcag cagtacggta gtagccccct cacctttggc 300 caggggacaa aagtcgaaat caagcgtacg gtggctgcac catctgtctt catcttcccg 360 ccatctgatg agcagttgaa atctggaact gcctctgttg tgtgcctgct gaataacttc 420 tatcccagag aggccaaagt acagtggaag gtggataacg ccctccaatc gggtaactcc 480 caggagagtg tcacagagca ggacagcaag gacagcacct acagcctcag cagcaccctg 540 acgctgagca aagcagacta cgagaaacac aaagtctacg cctgcgaagt cacccatcag 600 ggcctgagct cgcccgtcac aaagagcttc aacaggggag agtgt 645 <![CDATA[<210> 201]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> DNA CLC-564 P329GLALA IgG1重鏈]]> <![CDATA[<400> 201]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gttcgacgtt 300 ggtccgttcg actactgggg ccaaggaacc ctggtcaccg tctcgagtgc tagcaccaag 360 ggcccatcgg tcttccccct ggcaccctcc tccaagagca cctctggggg cacagcggcc 420 ctgggctgcc tggtcaagga ctacttcccc gaaccggtga cggtgtcgtg gaactcaggc 480 gccctgacca gcggcgtgca caccttcccg gctgtcctac agtcctcagg actctactcc 540 ctcagcagcg tggtgaccgt gccctccagc agcttgggca cccagaccta catctgcaac 600 gtgaatcaca agcccagcaa caccaaggtg gacaagaaag ttgagcccaa atcttgtgac 660 aaaactcaca catgcccacc gtgcccagca cctgaagctg cagggggacc gtcagtcttc 720 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 780 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 840 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatccc gggatgagct gaccaagaac 1080 caggtcagcc tgacctgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 202]]> <![CDATA[<211> 215]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> CLC-564 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 202]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro 85 90 95 Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala 100 105 110 Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser 115 120 125 Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140 Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser 145 150 155 160 Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175 Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190 Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys 195 200 205 Ser Phe Asn Arg Gly Glu Cys 210 215 <![CDATA[<210> 203]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> CLC-564 P329GLALA IgG1重鏈]]> <![CDATA[<400> 203]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Phe Asp Val Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 204]]> <![CDATA[<211> 639]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 17A9 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 204]]> tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60 acatgccaag gagacagcct cagaagttat tatgcaagct ggtaccagca gaagccagga 120 caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga 180 ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa 240 gatgaggctg actattactg taactcccgt gttatgcctc ataatcgcgt attcggcgga 300 gggaccaagc tgaccgtcct aggtcaaccc aaggctgccc ccagcgtgac cctgttcccc 360 cccagcagcg aggaactgca ggccaacaag gccaccctgg tctgcctgat cagcgacttc 420 tacccaggcg ccgtgaccgt ggcctggaag gccgacagca gccccgtgaa ggccggcgtg 480 gagaccacca cccccagcaa gcagagcaac aacaagtacg ccgccagcag ctacctgagc 540 ctgacccccg agcagtggaa gagccacagg tcctacagct gccaggtgac ccacgagggc 600 agcaccgtgg agaaaaccgt ggcccccacc gagtgcagc 639 <![CDATA[<210> 205]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 17A9 P329GLALA IgG1重鏈]]> <![CDATA[<400> 205]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgttttc 300 taccgtggtg gtgtttctat ggactactgg ggccaaggaa ccctggtcac cgtctcgagt 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 206]]> <![CDATA[<211> 213]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> 17A9 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 206]]> Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Arg Val Met Pro His Asn Arg 85 90 95 Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys Ala 100 105 110 Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln Ala 115 120 125 Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly Ala 130 135 140 Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly Val 145 150 155 160 Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala Ser 165 170 175 Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser Tyr 180 185 190 Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val Ala 195 200 205 Pro Thr Glu Cys Ser 210 <![CDATA[<210> 207]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工]]> <![CDATA[<220>]]> <![CDATA[<223> 17A9 P329GLALA IgG1重鏈]]> <![CDATA[<400> 207]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Phe Tyr Arg Gly Gly Val Ser Met Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 208]]> <![CDATA[<211> 663]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 人類OX40 His核苷酸序列]]> <![CDATA[<400> 208]]> ctgcactgcg tgggcgacac ctaccccagc aacgaccggt gctgccacga gtgcagaccc 60 ggcaacggca tggtgtcccg gtgcagccgg tcccagaaca ccgtgtgcag accttgcggc 120 cctggcttct acaacgacgt ggtgtccagc aagccctgca agccttgtac ctggtgcaac 180 ctgcggagcg gcagcgagcg gaagcagctg tgtaccgcca cccaggatac cgtgtgccgg 240 tgtagagccg gcacccagcc cctggacagc tacaaacccg gcgtggactg cgccccttgc 300 cctcctggcc acttcagccc tggcgacaac caggcctgca agccttggac caactgcacc 360 ctggccggca agcacaccct gcagcccgcc agcaatagca gcgacgccat ctgcgaggac 420 cgggatcctc ctgccaccca gcctcaggaa acccagggcc ctcccgccag acccatcacc 480 gtgcagccta cagaggcctg gcccagaacc agccaggggc ctagcaccag acccgtggaa 540 gtgcctggcg gcagagccgt cgacgaacag ttatattttc agggcggctc aggcctgaac 600 gacatcttcg aggcccagaa gatcgagtgg cacgaggctc gagctcacca ccatcaccat 660 cac 663 <![CDATA[<210> 209]]> <![CDATA[<211> 221]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 人類OX40 His]]> <![CDATA[<400> 209]]> Leu His Cys Val Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys His 1 5 10 15 Glu Cys Arg Pro Gly Asn Gly Met Val Ser Arg Cys Ser Arg Ser Gln 20 25 30 Asn Thr Val Cys Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val 35 40 45 Ser Ser Lys Pro Cys Lys Pro Cys Thr Trp Cys Asn Leu Arg Ser Gly 50 55 60 Ser Glu Arg Lys Gln Leu Cys Thr Ala Thr Gln Asp Thr Val Cys Arg 65 70 75 80 Cys Arg Ala Gly Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp 85 90 95 Cys Ala Pro Cys Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala 100 105 110 Cys Lys Pro Trp Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln 115 120 125 Pro Ala Ser Asn Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro 130 135 140 Ala Thr Gln Pro Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Ile Thr 145 150 155 160 Val Gln Pro Thr Glu Ala Trp Pro Arg Thr Ser Gln Gly Pro Ser Thr 165 170 175 Arg Pro Val Glu Val Pro Gly Gly Arg Ala Val Asp Glu Gln Leu Tyr 180 185 190 Phe Gln Gly Gly Ser Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile 195 200 205 Glu Trp His Glu Ala Arg Ala His His His His His His 210 215 220 <![CDATA[<210> 210]]> <![CDATA[<211> 681]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA鼠類OX40 His]]> <![CDATA[<400> 210]]> gtgaccgcca gacggctgaa ctgcgtgaag cacacctacc ccagcggcca caagtgctgc 60 agagagtgcc agcccggcca cggcatggtg tccagatgcg accacacacg ggacaccctg 120 tgccaccctt gcgagacagg cttctacaac gaggccgtga actacgatac ctgcaagcag 180 tgcacccagt gcaaccacag aagcggcagc gagctgaagc agaactgcac ccccacccag 240 gataccgtgt gcagatgcag acccggcacc cagcccagac aggacagcgg ctacaagctg 300 ggcgtggact gcgtgccctg ccctcctggc cacttcagcc ccggcaacaa ccaggcctgc 360 aagccctgga ccaactgcac cctgagcggc aagcagacca gacaccccgc cagcgacagc 420 ctggatgccg tgtgcgagga cagaagcctg ctggccaccc tgctgtggga gacacagcgg 480 cccaccttca gacccaccac cgtgcagagc accaccgtgt ggcccagaac cagcgagctg 540 cccagtcctc ctaccctcgt gacacctgag ggccccgtcg acgaacagtt atattttcag 600 ggcggctcag gcctgaacga catcttcgag gcccagaaga tcgagtggca cgaggctcga 660 gctcaccacc atcaccatca c 681 <![CDATA[<210> 211]]> <![CDATA[<211> 226]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類OX40 His]]> <![CDATA[<400> 211]]> Thr Ala Arg Arg Leu Asn Cys Val Lys His Thr Tyr Pro Ser Gly His 1 5 10 15 Lys Cys Cys Arg Glu Cys Gln Pro Gly His Gly Met Val Ser Arg Cys 20 25 30 Asp His Thr Arg Asp Thr Leu Cys His Pro Cys Glu Thr Gly Phe Tyr 35 40 45 Asn Glu Ala Val Asn Tyr Asp Thr Cys Lys Gln Cys Thr Gln Cys Asn 50 55 60 His Arg Ser Gly Ser Glu Leu Lys Gln Asn Cys Thr Pro Thr Gln Asp 65 70 75 80 Thr Val Cys Arg Cys Arg Pro Gly Thr Gln Pro Arg Gln Asp Ser Gly 85 90 95 Tyr Lys Leu Gly Val Asp Cys Val Pro Cys Pro Pro Gly His Phe Ser 100 105 110 Pro Gly Asn Asn Gln Ala Cys Lys Pro Trp Thr Asn Cys Thr Leu Ser 115 120 125 Gly Lys Gln Thr Arg His Pro Ala Ser Asp Ser Leu Asp Ala Val Cys 130 135 140 Glu Asp Arg Ser Leu Leu Ala Thr Leu Leu Trp Glu Thr Gln Arg Pro 145 150 155 160 Thr Phe Arg Pro Thr Thr Val Gln Ser Thr Thr Val Trp Pro Arg Thr 165 170 175 Ser Glu Leu Pro Ser Pro Pro Thr Leu Val Thr Pro Glu Gly Pro Val 180 185 190 Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Gly Leu Asn Asp Ile Phe 195 200 205 Glu Ala Gln Lys Ile Glu Trp His Glu Ala Arg Ala His His His His 210 215 220 His His 225 <![CDATA[<210> 212]]> <![CDATA[<211> 1317]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 核苷酸序列二聚人類OX40抗原Fc]]> <![CDATA[<400> 212]]> ctgcactgcg tgggcgacac ctaccccagc aacgaccggt gctgccacga gtgcagaccc 60 ggcaacggca tggtgtcccg gtgcagccgg tcccagaaca ccgtgtgcag accttgcggc 120 cctggcttct acaacgacgt ggtgtccagc aagccctgca agccttgtac ctggtgcaac 180 ctgcggagcg gcagcgagcg gaagcagctg tgtaccgcca cccaggatac cgtgtgccgg 240 tgtagagccg gcacccagcc cctggacagc tacaaacccg gcgtggactg cgccccttgc 300 cctcctggcc acttcagccc tggcgacaac caggcctgca agccttggac caactgcacc 360 ctggccggca agcacaccct gcagcccgcc agcaatagca gcgacgccat ctgcgaggac 420 cgggatcctc ctgccaccca gcctcaggaa acccagggcc ctcccgccag acccatcacc 480 gtgcagccta cagaggcctg gcccagaacc agccaggggc ctagcaccag acccgtggaa 540 gtgcctggcg gcagagccgt cgacgaacag ttatattttc agggcggctc acccaaatct 600 gcagacaaaa ctcacacatg cccaccgtgc ccagcacctg aactcctggg gggaccgtca 660 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 720 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 780 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 840 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 900 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 960 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 1020 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1080 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1140 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1200 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1260 agcctctccc tgtctccggg taaatccggc tacccatacg atgttccaga ttacgct 1317 <![CDATA[<210> 213]]> <![CDATA[<211> 439]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 二聚人類OX40抗原Fc]]> <![CDATA[<400> 213]]> Leu His Cys Val Gly Asp Thr Tyr Pro Ser Asn Asp Arg Cys Cys His 1 5 10 15 Glu Cys Arg Pro Gly Asn Gly Met Val Ser Arg Cys Ser Arg Ser Gln 20 25 30 Asn Thr Val Cys Arg Pro Cys Gly Pro Gly Phe Tyr Asn Asp Val Val 35 40 45 Ser Ser Lys Pro Cys Lys Pro Cys Thr Trp Cys Asn Leu Arg Ser Gly 50 55 60 Ser Glu Arg Lys Gln Leu Cys Thr Ala Thr Gln Asp Thr Val Cys Arg 65 70 75 80 Cys Arg Ala Gly Thr Gln Pro Leu Asp Ser Tyr Lys Pro Gly Val Asp 85 90 95 Cys Ala Pro Cys Pro Pro Gly His Phe Ser Pro Gly Asp Asn Gln Ala 100 105 110 Cys Lys Pro Trp Thr Asn Cys Thr Leu Ala Gly Lys His Thr Leu Gln 115 120 125 Pro Ala Ser Asn Ser Ser Asp Ala Ile Cys Glu Asp Arg Asp Pro Pro 130 135 140 Ala Thr Gln Pro Gln Glu Thr Gln Gly Pro Pro Ala Arg Pro Ile Thr 145 150 155 160 Val Gln Pro Thr Glu Ala Trp Pro Arg Thr Ser Gln Gly Pro Ser Thr 165 170 175 Arg Pro Val Glu Val Pro Gly Gly Arg Ala Val Asp Glu Gln Leu Tyr 180 185 190 Phe Gln Gly Gly Ser Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro 195 200 205 Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 210 215 220 Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 225 230 235 240 Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 245 250 255 Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 260 265 270 Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 275 280 285 Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 290 295 300 Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 305 310 315 320 Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 325 330 335 Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 340 345 350 Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 355 360 365 Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 370 375 380 Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 385 390 395 400 Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 405 410 415 Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys Ser Gly Tyr Pro 420 425 430 Tyr Asp Val Pro Asp Tyr Ala 435 <![CDATA[<210> 214]]> <![CDATA[<211> 2064]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (8B9) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 214]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttggatgg actactgggg ccaagggacc accgtgaccg tctcctcagc tagcaccaag 360 ggcccatccg tgttccctct ggccccttcc agcaagtcta cctctggcgg cacagccgct 420 ctgggctgcc tcgtgaagga ctacttcccc gagcctgtga cagtgtcctg gaactctggc 480 gccctgacat ccggcgtgca cacctttcca gctgtgctgc agtcctccgg cctgtactcc 540 ctgtcctccg tcgtgacagt gccctccagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agccctccaa caccaaggtg gacaagaagg tggaacccaa gtcctgcgac 660 aagacccaca cctgtccccc ttgtcctgcc cctgaagctg ctggcggccc tagcgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatctccc ggacccccga agtgacctgc 780 gtggtggtgg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagcct agagaggaac agtacaactc cacctaccgg 900 gtggtgtccg tgctgaccgt gctgcaccag gattggctga acggcaaaga gtacaagtgc 960 aaggtgtcca acaaggccct gggagccccc atcgaaaaga ccatctccaa ggccaagggc 1020 cagcctcgcg agcctcaggt gtacaccctg ccccctagca gagatgagct gaccaagaac 1080 caggtgtccc tgacctgtct cgtgaaaggc ttctacccct ccgatatcgc cgtggaatgg 1140 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggactccgac 1200 ggctcattct tcctgtactc taagctgaca gtggacaagt cccggtggca gcagggcaac 1260 gtgttctcct gctccgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg 1320 tccctgtctc ccgggggagg cggaggatct ggcggaggcg gatccggtgg tggcggatct 1380 gggggcggtg gatctgaggt gcagctgctg gaatctgggg gaggactggt gcagccaggc 1440 ggatctctga ggctgtcctg cgctgcttcc ggctttacct tctccagcca cgccatgagt 1500 tgggtgcgcc aggcacccgg aaaaggactg gaatgggtgt cagccatctg ggcctccggc 1560 gagcagtact acgccgatag cgtgaagggc cggttcacca tctctcggga taacagcaag 1620 aatactctgt acctgcagat gaactccctg cgcgctgaag ataccgctgt gtattactgc 1680 gccaagggct ggctgggcaa cttcgattac tggggccagg gaaccctcgt gactgtctcg 1740 agcgcttctg tggccgctcc ctccgtgttc atcttcccac cttccgacga gcagctgaag 1800 tccggcactg cctctgtcgt gtgcctgctg aacaacttct accctcggga agccaaggtg 1860 cagtggaaag tggataacgc cctgcagtcc ggcaactccc aggaatccgt gaccgagcag 1920 gactccaagg acagcaccta ctccctgagc agcaccctga ccctgtccaa ggccgactac 1980 gagaagcaca aggtgtacgc ctgtgaagtg acccaccagg gcctgtccag ccccgtgacc 2040 aagtccttca accggggcga gtgc 2064 <![CDATA[<210> 215]]> <![CDATA[<211> 642]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA VLCH1-Light chain 2 (28H1)]]> <![CDATA[<400> 215]]> gagatcgtgc tgacccagtc tcccggcacc ctgagcctga gccctggcga gagagccacc 60 ctgagctgca gagccagcca gagcgtgagc cggagctacc tggcctggta tcagcagaag 120 cccggccagg cccccagact gctgatcatc ggcgccagca cccgggccac cggcatcccc 180 gatagattca gcggcagcgg ctccggcacc gacttcaccc tgaccatcag ccggctggaa 240 cccgaggact tcgccgtgta ctactgccag cagggccagg tgatcccccc caccttcggc 300 cagggcacca aggtggaaat caagagctcc gctagcacca agggcccctc cgtgtttcct 360 ctggccccca gcagcaagag cacctctggc ggaacagccg ccctgggctg cctggtgaaa 420 gactacttcc ccgagcccgt gaccgtgtcc tggaactctg gcgccctgac cagcggcgtg 480 cacacctttc cagccgtgct gcagagcagc ggcctgtact ccctgagcag cgtggtgaca 540 gtgccctcca gcagcctggg cacccagacc tacatctgca acgtgaacca caagcccagc 600 aacaccaaag tggacaagaa ggtggaaccc aagagctgcg ac 642 <![CDATA[<210> 216]]> <![CDATA[<211> 688]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (8B9) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 216]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Trp Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly 435 440 445 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 450 455 460 Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 465 470 475 480 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 485 490 495 His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 500 505 510 Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val 515 520 525 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 530 535 540 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 545 550 555 560 Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly Thr Leu 565 570 575 Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe Ile Phe 580 585 590 Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys 595 600 605 Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val 610 615 620 Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln 625 630 635 640 Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser 645 650 655 Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His 660 665 670 Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 675 680 685 <![CDATA[<210> 217]]> <![CDATA[<211> 214]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> VLCH1-Light chain 2 (28H1)]]> <![CDATA[<400> 217]]> Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly 1 5 10 15 Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Arg Ser 20 25 30 Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu 35 40 45 Ile Ile Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser 50 55 60 Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu 65 70 75 80 Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Gln Val Ile Pro 85 90 95 Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Ser Ser Ala Ser 100 105 110 Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr 115 120 125 Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro 130 135 140 Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val 145 150 155 160 His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser 165 170 175 Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile 180 185 190 Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val 195 200 205 Glu Pro Lys Ser Cys Asp 210 <![CDATA[<210> 218]]> <![CDATA[<211> 2070]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (49B4) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 218]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catccgtgtt ccctctggcc ccttccagca agtctacctc tggcggcaca 420 gccgctctgg gctgcctcgt gaaggactac ttccccgagc ctgtgacagt gtcctggaac 480 tctggcgccc tgacatccgg cgtgcacacc tttccagctg tgctgcagtc ctccggcctg 540 tactccctgt cctccgtcgt gacagtgccc tccagctctc tgggcaccca gacctacatc 600 tgcaacgtga accacaagcc ctccaacacc aaggtggaca agaaggtgga acccaagtcc 660 tgcgacaaga cccacacctg tcccccttgt cctgcccctg aagctgctgg cggccctagc 720 gtgttcctgt tccccccaaa gcccaaggac accctgatga tctcccggac ccccgaagtg 780 acctgcgtgg tggtggatgt gtcccacgag gaccctgaag tgaagttcaa ttggtacgtg 840 gacggcgtgg aagtgcacaa tgccaagacc aagcctagag aggaacagta caactccacc 900 taccgggtgg tgtccgtgct gaccgtgctg caccaggatt ggctgaacgg caaagagtac 960 aagtgcaagg tgtccaacaa ggccctggga gcccccatcg aaaagaccat ctccaaggcc 1020 aagggccagc ctcgcgagcc tcaggtgtac accctgcccc ctagcagaga tgagctgacc 1080 aagaaccagg tgtccctgac ctgtctcgtg aaaggcttct acccctccga tatcgccgtg 1140 gaatgggaga gcaacggcca gcccgagaac aactacaaga ccaccccccc tgtgctggac 1200 tccgacggct cattcttcct gtactctaag ctgacagtgg acaagtcccg gtggcagcag 1260 ggcaacgtgt tctcctgctc cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgtccc tgtctcccgg gggaggcgga ggatctggcg gaggcggatc cggtggtggc 1380 ggatctgggg gcggtggatc tgaggtgcag ctgctggaat ctgggggagg actggtgcag 1440 ccaggcggat ctctgaggct gtcctgcgct gcttccggct ttaccttctc cagccacgcc 1500 atgagttggg tgcgccaggc acccggaaaa ggactggaat gggtgtcagc catctgggcc 1560 tccggcgagc agtactacgc cgatagcgtg aagggccggt tcaccatctc tcgggataac 1620 agcaagaata ctctgtacct gcagatgaac tccctgcgcg ctgaagatac cgctgtgtat 1680 tactgcgcca agggctggct gggcaacttc gattactggg gccagggaac cctcgtgact 1740 gtctcgagcg cttctgtggc cgctccctcc gtgttcatct tcccaccttc cgacgagcag 1800 ctgaagtccg gcactgcctc tgtcgtgtgc ctgctgaaca acttctaccc tcgggaagcc 1860 aaggtgcagt ggaaagtgga taacgccctg cagtccggca actcccagga atccgtgacc 1920 gagcaggact ccaaggacag cacctactcc ctgagcagca ccctgaccct gtccaaggcc 1980 gactacgaga agcacaaggt gtacgcctgt gaagtgaccc accagggcct gtccagcccc 2040 gtgaccaagt ccttcaaccg gggcgagtgc 2070 <![CDATA[<210> 219]]> <![CDATA[<211> 690]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 219]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp 515 520 525 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 530 535 540 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 545 550 555 560 Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly 565 570 575 Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe 580 585 590 Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val 595 600 605 Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp 610 615 620 Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr 625 630 635 640 Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr 645 650 655 Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val 660 665 670 Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly 675 680 685 Glu Cys 690 <![CDATA[<210> 220]]> <![CDATA[<211> 2064]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (1G4) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 220]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttctatgg actactgggg ccaagggacc accgtgaccg tctcctcagc tagcaccaag 360 ggcccatccg tgttccctct ggccccttcc agcaagtcta cctctggcgg cacagccgct 420 ctgggctgcc tcgtgaagga ctacttcccc gagcctgtga cagtgtcctg gaactctggc 480 gccctgacat ccggcgtgca cacctttcca gctgtgctgc agtcctccgg cctgtactcc 540 ctgtcctccg tcgtgacagt gccctccagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agccctccaa caccaaggtg gacaagaagg tggaacccaa gtcctgcgac 660 aagacccaca cctgtccccc ttgtcctgcc cctgaagctg ctggcggccc tagcgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatctccc ggacccccga agtgacctgc 780 gtggtggtgg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagcct agagaggaac agtacaactc cacctaccgg 900 gtggtgtccg tgctgaccgt gctgcaccag gattggctga acggcaaaga gtacaagtgc 960 aaggtgtcca acaaggccct gggagccccc atcgaaaaga ccatctccaa ggccaagggc 1020 cagcctcgcg agcctcaggt gtacaccctg ccccctagca gagatgagct gaccaagaac 1080 caggtgtccc tgacctgtct cgtgaaaggc ttctacccct ccgatatcgc cgtggaatgg 1140 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggactccgac 1200 ggctcattct tcctgtactc taagctgaca gtggacaagt cccggtggca gcagggcaac 1260 gtgttctcct gctccgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg 1320 tccctgtctc ccgggggagg cggaggatct ggcggaggcg gatccggtgg tggcggatct 1380 gggggcggtg gatctgaggt gcagctgctg gaatctgggg gaggactggt gcagccaggc 1440 ggatctctga ggctgtcctg cgctgcttcc ggctttacct tctccagcca cgccatgagt 1500 tgggtgcgcc aggcacccgg aaaaggactg gaatgggtgt cagccatctg ggcctccggc 1560 gagcagtact acgccgatag cgtgaagggc cggttcacca tctctcggga taacagcaag 1620 aatactctgt acctgcagat gaactccctg cgcgctgaag ataccgctgt gtattactgc 1680 gccaagggct ggctgggcaa cttcgattac tggggccagg gaaccctcgt gactgtctcg 1740 agcgcttctg tggccgctcc ctccgtgttc atcttcccac cttccgacga gcagctgaag 1800 tccggcactg cctctgtcgt gtgcctgctg aacaacttct accctcggga agccaaggtg 1860 cagtggaaag tggataacgc cctgcagtcc ggcaactccc aggaatccgt gaccgagcag 1920 gactccaagg acagcaccta ctccctgagc agcaccctga ccctgtccaa ggccgactac 1980 gagaagcaca aggtgtacgc ctgtgaagtg acccaccagg gcctgtccag ccccgtgacc 2040 aagtccttca accggggcga gtgc 2064 <![CDATA[<210> 221]]> <![CDATA[<211> 688]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (1G4) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 221]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Ser Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly 435 440 445 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 450 455 460 Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 465 470 475 480 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 485 490 495 His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 500 505 510 Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val 515 520 525 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 530 535 540 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 545 550 555 560 Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly Thr Leu 565 570 575 Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe Ile Phe 580 585 590 Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys 595 600 605 Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val 610 615 620 Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln 625 630 635 640 Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser 645 650 655 Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His 660 665 670 Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 675 680 685 <![CDATA[<210> 222]]> <![CDATA[<211> 2082]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (20B7) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 222]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagttaac 300 tacccgtact cttactgggg tgacttcgac tactggggcc aagggaccac cgtgaccgtc 360 tcctcagcta gcaccaaggg cccatccgtg ttccctctgg ccccttccag caagtctacc 420 tctggcggca cagccgctct gggctgcctc gtgaaggact acttccccga gcctgtgaca 480 gtgtcctgga actctggcgc cctgacatcc ggcgtgcaca cctttccagc tgtgctgcag 540 tcctccggcc tgtactccct gtcctccgtc gtgacagtgc cctccagctc tctgggcacc 600 cagacctaca tctgcaacgt gaaccacaag ccctccaaca ccaaggtgga caagaaggtg 660 gaacccaagt cctgcgacaa gacccacacc tgtccccctt gtcctgcccc tgaagctgct 720 ggcggcccta gcgtgttcct gttcccccca aagcccaagg acaccctgat gatctcccgg 780 acccccgaag tgacctgcgt ggtggtggat gtgtcccacg aggaccctga agtgaagttc 840 aattggtacg tggacggcgt ggaagtgcac aatgccaaga ccaagcctag agaggaacag 900 tacaactcca cctaccgggt ggtgtccgtg ctgaccgtgc tgcaccagga ttggctgaac 960 ggcaaagagt acaagtgcaa ggtgtccaac aaggccctgg gagcccccat cgaaaagacc 1020 atctccaagg ccaagggcca gcctcgcgag cctcaggtgt acaccctgcc ccctagcaga 1080 gatgagctga ccaagaacca ggtgtccctg acctgtctcg tgaaaggctt ctacccctcc 1140 gatatcgccg tggaatggga gagcaacggc cagcccgaga acaactacaa gaccaccccc 1200 cctgtgctgg actccgacgg ctcattcttc ctgtactcta agctgacagt ggacaagtcc 1260 cggtggcagc agggcaacgt gttctcctgc tccgtgatgc acgaggccct gcacaaccac 1320 tacacccaga agtccctgtc cctgtctccc gggggaggcg gaggatctgg cggaggcgga 1380 tccggtggtg gcggatctgg gggcggtgga tctgaggtgc agctgctgga atctggggga 1440 ggactggtgc agccaggcgg atctctgagg ctgtcctgcg ctgcttccgg ctttaccttc 1500 tccagccacg ccatgagttg ggtgcgccag gcacccggaa aaggactgga atgggtgtca 1560 gccatctggg cctccggcga gcagtactac gccgatagcg tgaagggccg gttcaccatc 1620 tctcgggata acagcaagaa tactctgtac ctgcagatga actccctgcg cgctgaagat 1680 accgctgtgt attactgcgc caagggctgg ctgggcaact tcgattactg gggccaggga 1740 accctcgtga ctgtctcgag cgcttctgtg gccgctccct ccgtgttcat cttcccacct 1800 tccgacgagc agctgaagtc cggcactgcc tctgtcgtgt gcctgctgaa caacttctac 1860 cctcgggaag ccaaggtgca gtggaaagtg gataacgccc tgcagtccgg caactcccag 1920 gaatccgtga ccgagcagga ctccaaggac agcacctact ccctgagcag caccctgacc 1980 ctgtccaagg ccgactacga gaagcacaag gtgtacgcct gtgaagtgac ccaccagggc 2040 ctgtccagcc ccgtgaccaa gtccttcaac cggggcgagt gc 2082 <![CDATA[<210> 223]]> <![CDATA[<211> 694]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (20B7) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 223]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Asn Tyr Pro Tyr Ser Tyr Trp Gly Asp Phe Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val 355 360 365 Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly 450 455 460 Gly Ser Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly 465 470 475 480 Gly Leu Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser 485 490 495 Gly Phe Thr Phe Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro 500 505 510 Gly Lys Gly Leu Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln 515 520 525 Tyr Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn 530 535 540 Ser Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp 545 550 555 560 Thr Ala Val Tyr Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr 565 570 575 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala 580 585 590 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 595 600 605 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 610 615 620 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 625 630 635 640 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 645 650 655 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 660 665 670 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 675 680 685 Phe Asn Arg Gly Glu Cys 690 <![CDATA[<210> 224]]> <![CDATA[<211> 2064]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-563) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 224]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcttgacgtt 300 ggtgctttcg actactgggg ccaaggagcc ctggtcaccg tctcgagtgc tagcaccaag 360 ggcccatccg tgttccctct ggccccttcc agcaagtcta cctctggcgg cacagccgct 420 ctgggctgcc tcgtgaagga ctacttcccc gagcctgtga cagtgtcctg gaactctggc 480 gccctgacat ccggcgtgca cacctttcca gctgtgctgc agtcctccgg cctgtactcc 540 ctgtcctccg tcgtgacagt gccctccagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agccctccaa caccaaggtg gacaagaagg tggaacccaa gtcctgcgac 660 aagacccaca cctgtccccc ttgtcctgcc cctgaagctg ctggcggccc tagcgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatctccc ggacccccga agtgacctgc 780 gtggtggtgg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagcct agagaggaac agtacaactc cacctaccgg 900 gtggtgtccg tgctgaccgt gctgcaccag gattggctga acggcaaaga gtacaagtgc 960 aaggtgtcca acaaggccct gggagccccc atcgaaaaga ccatctccaa ggccaagggc 1020 cagcctcgcg agcctcaggt gtacaccctg ccccctagca gagatgagct gaccaagaac 1080 caggtgtccc tgacctgtct cgtgaaaggc ttctacccct ccgatatcgc cgtggaatgg 1140 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggactccgac 1200 ggctcattct tcctgtactc taagctgaca gtggacaagt cccggtggca gcagggcaac 1260 gtgttctcct gctccgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg 1320 tccctgtctc ccgggggagg cggaggatct ggcggaggcg gatccggtgg tggcggatct 1380 gggggcggtg gatctgaggt gcagctgctg gaatctgggg gaggactggt gcagccaggc 1440 ggatctctga ggctgtcctg cgctgcttcc ggctttacct tctccagcca cgccatgagt 1500 tgggtgcgcc aggcacccgg aaaaggactg gaatgggtgt cagccatctg ggcctccggc 1560 gagcagtact acgccgatag cgtgaagggc cggttcacca tctctcggga taacagcaag 1620 aatactctgt acctgcagat gaactccctg cgcgctgaag ataccgctgt gtattactgc 1680 gccaagggct ggctgggcaa cttcgattac tggggccagg gaaccctcgt gactgtctcg 1740 agcgcttctg tggccgctcc ctccgtgttc atcttcccac cttccgacga gcagctgaag 1800 tccggcactg cctctgtcgt gtgcctgctg aacaacttct accctcggga agccaaggtg 1860 cagtggaaag tggataacgc cctgcagtcc ggcaactccc aggaatccgt gaccgagcag 1920 gactccaagg acagcaccta ctccctgagc agcaccctga ccctgtccaa ggccgactac 1980 gagaagcaca aggtgtacgc ctgtgaagtg acccaccagg gcctgtccag ccccgtgacc 2040 aagtccttca accggggcga gtgc 2064 <![CDATA[<210> 225]]> <![CDATA[<211> 688]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (CLC-563) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 225]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Leu Asp Val Gly Ala Phe Asp Tyr Trp Gly Gln Gly Ala Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly 435 440 445 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 450 455 460 Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 465 470 475 480 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 485 490 495 His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 500 505 510 Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val 515 520 525 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 530 535 540 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 545 550 555 560 Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly Thr Leu 565 570 575 Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe Ile Phe 580 585 590 Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys 595 600 605 Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val 610 615 620 Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln 625 630 635 640 Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser 645 650 655 Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His 660 665 670 Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 675 680 685 <![CDATA[<210> 226]]> <![CDATA[<211> 2064]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-564) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 226]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gttcgacgtt 300 ggtccgttcg actactgggg ccaaggaacc ctggtcaccg tctcgagtgc tagcaccaag 360 ggcccatccg tgttccctct ggccccttcc agcaagtcta cctctggcgg cacagccgct 420 ctgggctgcc tcgtgaagga ctacttcccc gagcctgtga cagtgtcctg gaactctggc 480 gccctgacat ccggcgtgca cacctttcca gctgtgctgc agtcctccgg cctgtactcc 540 ctgtcctccg tcgtgacagt gccctccagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agccctccaa caccaaggtg gacaagaagg tggaacccaa gtcctgcgac 660 aagacccaca cctgtccccc ttgtcctgcc cctgaagctg ctggcggccc tagcgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatctccc ggacccccga agtgacctgc 780 gtggtggtgg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagcct agagaggaac agtacaactc cacctaccgg 900 gtggtgtccg tgctgaccgt gctgcaccag gattggctga acggcaaaga gtacaagtgc 960 aaggtgtcca acaaggccct gggagccccc atcgaaaaga ccatctccaa ggccaagggc 1020 cagcctcgcg agcctcaggt gtacaccctg ccccctagca gagatgagct gaccaagaac 1080 caggtgtccc tgacctgtct cgtgaaaggc ttctacccct ccgatatcgc cgtggaatgg 1140 gagagcaacg gccagcccga gaacaactac aagaccaccc cccctgtgct ggactccgac 1200 ggctcattct tcctgtactc taagctgaca gtggacaagt cccggtggca gcagggcaac 1260 gtgttctcct gctccgtgat gcacgaggcc ctgcacaacc actacaccca gaagtccctg 1320 tccctgtctc ccgggggagg cggaggatct ggcggaggcg gatccggtgg tggcggatct 1380 gggggcggtg gatctgaggt gcagctgctg gaatctgggg gaggactggt gcagccaggc 1440 ggatctctga ggctgtcctg cgctgcttcc ggctttacct tctccagcca cgccatgagt 1500 tgggtgcgcc aggcacccgg aaaaggactg gaatgggtgt cagccatctg ggcctccggc 1560 gagcagtact acgccgatag cgtgaagggc cggttcacca tctctcggga taacagcaag 1620 aatactctgt acctgcagat gaactccctg cgcgctgaag ataccgctgt gtattactgc 1680 gccaagggct ggctgggcaa cttcgattac tggggccagg gaaccctcgt gactgtctcg 1740 agcgcttctg tggccgctcc ctccgtgttc atcttcccac cttccgacga gcagctgaag 1800 tccggcactg cctctgtcgt gtgcctgctg aacaacttct accctcggga agccaaggtg 1860 cagtggaaag tggataacgc cctgcagtcc ggcaactccc aggaatccgt gaccgagcag 1920 gactccaagg acagcaccta ctccctgagc agcaccctga ccctgtccaa ggccgactac 1980 gagaagcaca aggtgtacgc ctgtgaagtg acccaccagg gcctgtccag ccccgtgacc 2040 aagtccttca accggggcga gtgc 2064 <![CDATA[<210> 227]]> <![CDATA[<211> 688]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (CLC-564) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 227]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Phe Asp Val Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly Gly Gly 435 440 445 Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly 450 455 460 Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly 465 470 475 480 Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser 485 490 495 His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp 500 505 510 Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val 515 520 525 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 530 535 540 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 545 550 555 560 Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly Thr Leu 565 570 575 Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe Ile Phe 580 585 590 Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys 595 600 605 Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val 610 615 620 Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln 625 630 635 640 Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser 645 650 655 Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His 660 665 670 Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 675 680 685 <![CDATA[<210> 228]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (28H1) VHCL-重鏈臼]]> <![CDATA[<400> 228]]> gaagtgcagc tgctggaatc cggcggaggc ctggtgcagc ctggcggatc tctgagactg 60 tcctgcgccg cctccggctt caccttctcc tcccacgcca tgtcctgggt ccgacaggct 120 cctggcaaag gcctggaatg ggtgtccgcc atctgggcct ccggcgagca gtactacgcc 180 gactctgtga agggccggtt caccatctcc cgggacaact ccaagaacac cctgtacctg 240 cagatgaact ccctgcgggc cgaggacacc gccgtgtact actgtgccaa gggctggctg 300 ggcaacttcg actactgggg acagggcacc ctggtcaccg tgtccagcgc tagcgtggcc 360 gctcccagcg tgttcatctt cccacccagc gacgagcagc tgaagtccgg cacagccagc 420 gtggtgtgcc tgctgaacaa cttctacccc cgcgaggcca aggtgcagtg gaaggtggac 480 aacgccctgc agagcggcaa cagccaggaa tccgtgaccg agcaggacag caaggactcc 540 acctacagcc tgagcagcac cctgaccctg agcaaggccg actacgagaa gcacaaggtg 600 tacgcctgcg aagtgaccca ccagggcctg tccagccccg tgaccaagag cttcaaccgg 660 ggcgagtgcg acaagaccca cacctgtccc ccttgccctg cccctgaagc tgctggtggc 720 ccttccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc 780 gaagtgacct gcgtggtggt cgatgtgtcc cacgaggacc ctgaagtgaa gttcaattgg 840 tacgtggacg gcgtggaagt gcacaatgcc aagaccaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtgcaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctctcgtgc gcagtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctcgtg agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 229]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (28H1) VHCL-重鏈臼]]> <![CDATA[<400> 229]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser His 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp Ser Val Lys 50 55 60 Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr Leu 65 70 75 80 Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala 85 90 95 Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe Ile Phe Pro 115 120 125 Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu 130 135 140 Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp 145 150 155 160 Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp 165 170 175 Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys 180 185 190 Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln 195 200 205 Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 230]]> <![CDATA[<211> 1343]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (49B4) VHCH1-重鏈臼]]> <![CDATA[<400> 230]]> aggtgcaatt ggtgcagtct ggggctgagg tgaagaagcc tgggtcctcg gtgaaggtct 60 cctgcaaggc ctccggaggc acattcagca gctacgctat aagctgggtg cgacaggccc 120 ctggacaagg gctcgagtgg atgggaggga tcatccctat ctttggtaca gcaaactacg 180 cacagaagtt ccagggcagg gtcaccatta ctgcagacaa atccacgagc acagcctaca 240 tggagctgag cagcctgaga tctgaggaca ccgccgtgta ttactgtgcg agagaatact 300 accgtggtcc gtacgactac tggggccaag ggaccaccgt gaccgtctcc tcagctagca 360 ccaagggccc tagcgtgttc cctctggccc ctagcagcaa gagcacaagt ggaggaacag 420 ccgccctggg ctgcctggtc aaggactact tccccgagcc cgtgaccgtg tcctggaatt 480 ctggcgccct gacaagcggc gtgcacacat ttccagccgt gctgcagagc agcggcctgt 540 actctctgag cagcgtcgtg accgtgccct ctagctctct gggcacccag acctacatct 600 gcaacgtgaa ccacaagccc agcaacacca aagtggacaa gaaggtggaa cccaagagct 660 gcgacaagac ccacacctgt cccccttgcc ctgcccctga agctgctggt ggcccttccg 720 tgttcctgtt ccccccaaag cccaaggaca ccctgatgat cagccggacc cccgaagtga 780 cctgcgtggt ggtcgatgtg tcccacgagg accctgaagt gaagttcaat tggtacgtgg 840 acggcgtgga agtgcacaat gccaagacca agccgcggga ggagcagtac aacagcacgt 900 accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc aaggagtaca 960 agtgcaaggt ctccaacaaa gccctcggcg cccccatcga gaaaaccatc tccaaagcca 1020 aagggcagcc ccgagaacca caggtgtaca ccctgccccc atgccgggat gagctgacca 1080 agaaccaggt cagcctgtgg tgcctggtca aaggcttcta tcccagcgac atcgccgtgg 1140 agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc gtgctggact 1200 ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg tggcagcagg 1260 ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac acgcagaaga 1320 gcctctccct gtctccgggt aaa 1343 <![CDATA[<210> 231]]> <![CDATA[<211> 448]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1-重鏈臼]]> <![CDATA[<400> 231]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 232]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (1G4) VHCH1-重鏈臼]]> <![CDATA[<400> 232]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttctatgg actactgggg ccaagggacc accgtgaccg tctcctcagc tagcaccaag 360 ggccctagcg tgttccctct ggcccctagc agcaagagca caagtggagg aacagccgcc 420 ctgggctgcc tggtcaagga ctacttcccc gagcccgtga ccgtgtcctg gaattctggc 480 gccctgacaa gcggcgtgca cacatttcca gccgtgctgc agagcagcgg cctgtactct 540 ctgagcagcg tcgtgaccgt gccctctagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agcccagcaa caccaaagtg gacaagaagg tggaacccaa gagctgcgac 660 aagacccaca cctgtccccc ttgccctgcc cctgaagctg ctggtggccc ttccgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 780 gtggtggtcg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatgcc gggatgagct gaccaagaac 1080 caggtcagcc tgtggtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 233]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (1G4) VHCH1-重鏈臼]]> <![CDATA[<400> 233]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Ser Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 234]]> <![CDATA[<211> 1356]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (20B7) VHCH1-重鏈臼]]> <![CDATA[<400> 234]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagttaac 300 tacccgtact cttactgggg tgacttcgac tactggggcc aagggaccac cgtgaccgtc 360 tcctcagcta gcaccaaggg ccctagcgtg ttccctctgg cccctagcag caagagcaca 420 agtggaggaa cagccgccct gggctgcctg gtcaaggact acttccccga gcccgtgacc 480 gtgtcctgga attctggcgc cctgacaagc ggcgtgcaca catttccagc cgtgctgcag 540 agcagcggcc tgtactctct gagcagcgtc gtgaccgtgc cctctagctc tctgggcacc 600 cagacctaca tctgcaacgt gaaccacaag cccagcaaca ccaaagtgga caagaaggtg 660 gaacccaaga gctgcgacaa gacccacacc tgtccccctt gccctgcccc tgaagctgct 720 ggtggccctt ccgtgttcct gttcccccca aagcccaagg acaccctgat gatcagccgg 780 acccccgaag tgacctgcgt ggtggtcgat gtgtcccacg aggaccctga agtgaagttc 840 aattggtacg tggacggcgt ggaagtgcac aatgccaaga ccaagccgcg ggaggagcag 900 tacaacagca cgtaccgtgt ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat 960 ggcaaggagt acaagtgcaa ggtctccaac aaagccctcg gcgcccccat cgagaaaacc 1020 atctccaaag ccaaagggca gccccgagaa ccacaggtgt acaccctgcc cccatgccgg 1080 gatgagctga ccaagaacca ggtcagcctg tggtgcctgg tcaaaggctt ctatcccagc 1140 gacatcgccg tggagtggga gagcaatggg cagccggaga acaactacaa gaccacgcct 1200 cccgtgctgg actccgacgg ctccttcttc ctctacagca agctcaccgt ggacaagagc 1260 aggtggcagc aggggaacgt cttctcatgc tccgtgatgc atgaggctct gcacaaccac 1320 tacacgcaga agagcctctc cctgtctccg ggtaaa 1356 <![CDATA[<210> 235]]> <![CDATA[<211> 452]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (20B7) VHCH1-重鏈臼]]> <![CDATA[<400> 235]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Val Asn Tyr Pro Tyr Ser Tyr Trp Gly Asp Phe Asp Tyr Trp 100 105 110 Gly Gln Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro 115 120 125 Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr 130 135 140 Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr 145 150 155 160 Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro 165 170 175 Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr 180 185 190 Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn 195 200 205 His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser 210 215 220 Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala 225 230 235 240 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 245 250 255 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 260 265 270 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 275 280 285 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 290 295 300 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 305 310 315 320 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro 325 330 335 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 340 345 350 Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val 355 360 365 Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 370 375 380 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 385 390 395 400 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 405 410 415 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 420 425 430 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 435 440 445 Ser Pro Gly Lys 450 <![CDATA[<210> 236]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-563) VHCH1-重鏈臼]]> <![CDATA[<400> 236]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcttgacgtt 300 ggtgctttcg actactgggg ccaaggagcc ctggtcaccg tctcgagtgc tagcaccaag 360 ggccctagcg tgttccctct ggcccctagc agcaagagca caagtggagg aacagccgcc 420 ctgggctgcc tggtcaagga ctacttcccc gagcccgtga ccgtgtcctg gaattctggc 480 gccctgacaa gcggcgtgca cacatttcca gccgtgctgc agagcagcgg cctgtactct 540 ctgagcagcg tcgtgaccgt gccctctagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agcccagcaa caccaaagtg gacaagaagg tggaacccaa gagctgcgac 660 aagacccaca cctgtccccc ttgccctgcc cctgaagctg ctggtggccc ttccgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 780 gtggtggtcg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatgcc gggatgagct gaccaagaac 1080 caggtcagcc tgtggtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 237]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (CLC-563) VHCH1-重鏈臼]]> <![CDATA[<400> 237]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Leu Asp Val Gly Ala Phe Asp Tyr Trp Gly Gln Gly Ala Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 238]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (CLC-564) VHCH1-重鏈臼]]> <![CDATA[<400> 238]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gttcgacgtt 300 ggtccgttcg actactgggg ccaaggaacc ctggtcaccg tctcgagtgc tagcaccaag 360 ggccctagcg tgttccctct ggcccctagc agcaagagca caagtggagg aacagccgcc 420 ctgggctgcc tggtcaagga ctacttcccc gagcccgtga ccgtgtcctg gaattctggc 480 gccctgacaa gcggcgtgca cacatttcca gccgtgctgc agagcagcgg cctgtactct 540 ctgagcagcg tcgtgaccgt gccctctagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agcccagcaa caccaaagtg gacaagaagg tggaacccaa gagctgcgac 660 aagacccaca cctgtccccc ttgccctgcc cctgaagctg ctggtggccc ttccgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 780 gtggtggtcg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatgcc gggatgagct gaccaagaac 1080 caggtcagcc tgtggtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 239]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (CLC-564) VHCH1-重鏈臼]]> <![CDATA[<400> 239]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Phe Asp Val Gly Pro Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 240]]> <![CDATA[<211> 163]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 獼猴]]> <![CDATA[<400> 240]]> Leu Gln Asp Leu Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Ser Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Lys Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Ile Ser Gly Tyr His Cys Leu Gly Ala Glu Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp 85 90 95 Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro 100 105 110 Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr 115 120 125 Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro 130 135 140 Gly Ala Ser Ser Ala Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His 145 150 155 160 Ser Pro Gln <![CDATA[<210> 241]]> <![CDATA[<211> 164]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 小家鼠]]> <![CDATA[<400> 241]]> Val Gln Asn Ser Cys Asp Asn Cys Gln Pro Gly Thr Phe Cys Arg Lys 1 5 10 15 Tyr Asn Pro Val Cys Lys Ser Cys Pro Pro Ser Thr Phe Ser Ser Ile 20 25 30 Gly Gly Gln Pro Asn Cys Asn Ile Cys Arg Val Cys Ala Gly Tyr Phe 35 40 45 Arg Phe Lys Lys Phe Cys Ser Ser Thr His Asn Ala Glu Cys Glu Cys 50 55 60 Ile Glu Gly Phe His Cys Leu Gly Pro Gln Cys Thr Arg Cys Glu Lys 65 70 75 80 Asp Cys Arg Pro Gly Gln Glu Leu Thr Lys Gln Gly Cys Lys Thr Cys 85 90 95 Ser Leu Gly Thr Phe Asn Asp Gln Asn Gly Thr Gly Val Cys Arg Pro 100 105 110 Trp Thr Asn Cys Ser Leu Asp Gly Arg Ser Val Leu Lys Thr Gly Thr 115 120 125 Thr Glu Lys Asp Val Val Cys Gly Pro Pro Val Val Ser Phe Ser Pro 130 135 140 Ser Thr Thr Ile Ser Val Thr Pro Glu Gly Gly Pro Gly Gly His Ser 145 150 155 160 Leu Gln Val Leu <![CDATA[<210> 242]]> <![CDATA[<211> 1266]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 人類4-1BB抗原Fc杵鏈]]> <![CDATA[<400> 242]]> ctgcaggacc cctgcagcaa ctgccctgcc ggcaccttct gcgacaacaa ccggaaccag 60 atctgcagcc cctgcccccc caacagcttc agctctgccg gcggacagcg gacctgcgac 120 atctgcagac agtgcaaggg cgtgttcaga acccggaaag agtgcagcag caccagcaac 180 gccgagtgcg actgcacccc cggcttccat tgtctgggag ccggctgcag catgtgcgag 240 caggactgca agcagggcca ggaactgacc aagaagggct gcaaggactg ctgcttcggc 300 accttcaacg accagaagcg gggcatctgc cggccctgga ccaactgtag cctggacggc 360 aagagcgtgc tggtcaacgg caccaaagaa cgggacgtcg tgtgcggccc cagccctgct 420 gatctgtctc ctggggccag cagcgtgacc cctcctgccc ctgccagaga gcctggccac 480 tctcctcagg tcgacgaaca gttatatttt cagggcggct cacccaaatc tgcagacaaa 540 actcacacat gcccaccgtg cccagcacct gaactcctgg ggggaccgtc agtcttcctc 600 ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 660 gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 720 gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg 780 gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 840 gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag 900 ccccgagaac cacaggtgta caccctgccc ccatgccggg atgagctgac caagaaccag 960 gtcagcctgt ggtgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 1020 agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc 1080 tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc 1140 ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc 1200 ctgtctccgg gtaaatccgg aggcctgaac gacatcttcg aggcccagaa gattgaatgg 1260 cacgag 1266 <![CDATA[<210> 243]]> <![CDATA[<211> 1266]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA cynomolgus 4-1BB抗原Fc杵鏈]]> <![CDATA[<400> 243]]> ttgcaggatc tgtgtagtaa ctgcccagct ggtacattct gtgataataa caggagtcag 60 atttgcagtc cctgtcctcc aaatagtttc tccagcgcag gtggacaaag gacctgtgac 120 atatgcaggc agtgtaaagg tgttttcaag accaggaagg agtgttcctc caccagcaat 180 gcagagtgtg actgcatttc agggtatcac tgcctggggg cagagtgcag catgtgtgaa 240 caggattgta aacaaggtca agaattgaca aaaaaaggtt gtaaagactg ttgctttggg 300 acatttaatg accagaaacg tggcatctgt cgcccctgga caaactgttc tttggatgga 360 aagtctgtgc ttgtgaatgg gacgaaggag agggacgtgg tctgcggacc atctccagcc 420 gacctctctc caggagcatc ctctgcgacc ccgcctgccc ctgcgagaga gccaggacac 480 tctccgcagg tcgacgaaca gttatatttt cagggcggct cacccaaatc tgcagacaaa 540 actcacacat gcccaccgtg cccagcacct gaactcctgg ggggaccgtc agtcttcctc 600 ttccccccaa aacccaagga caccctcatg atctcccgga cccctgaggt cacatgcgtg 660 gtggtggacg tgagccacga agaccctgag gtcaagttca actggtacgt ggacggcgtg 720 gaggtgcata atgccaagac aaagccgcgg gaggagcagt acaacagcac gtaccgtgtg 780 gtcagcgtcc tcaccgtcct gcaccaggac tggctgaatg gcaaggagta caagtgcaag 840 gtctccaaca aagccctccc agcccccatc gagaaaacca tctccaaagc caaagggcag 900 ccccgagaac cacaggtgta caccctgccc ccatgccggg atgagctgac caagaaccag 960 gtcagcctgt ggtgcctggt caaaggcttc tatcccagcg acatcgccgt ggagtgggag 1020 agcaatgggc agccggagaa caactacaag accacgcctc ccgtgctgga ctccgacggc 1080 tccttcttcc tctacagcaa gctcaccgtg gacaagagca ggtggcagca ggggaacgtc 1140 ttctcatgct ccgtgatgca tgaggctctg cacaaccact acacgcagaa gagcctctcc 1200 ctgtctccgg gtaaatccgg aggcctgaac gacatcttcg aggcccagaa gattgaatgg 1260 cacgag 1266 <![CDATA[<210> 244]]> <![CDATA[<211> 1269]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA鼠類4-1BB抗原Fc杵鏈]]> <![CDATA[<400> 244]]> gtgcagaaca gctgcgacaa ctgccagccc ggcaccttct gccggaagta caaccccgtg 60 tgcaagagct gcccccccag caccttcagc agcatcggcg gccagcccaa ctgcaacatc 120 tgcagagtgt gcgccggcta cttccggttc aagaagttct gcagcagcac ccacaacgcc 180 gagtgcgagt gcatcgaggg cttccactgc ctgggccccc agtgcaccag atgcgagaag 240 gactgcagac ccggccagga actgaccaag cagggctgta agacctgcag cctgggcacc 300 ttcaacgacc agaacgggac cggcgtgtgc cggccttgga ccaattgcag cctggacggg 360 agaagcgtgc tgaaaaccgg caccaccgag aaggacgtcg tgtgcggccc tcccgtggtg 420 tccttcagcc ctagcaccac catcagcgtg acccctgaag gcggccctgg cggacactct 480 ctgcaggtcc tggtcgacga acagttatat tttcagggcg gctcacccaa atctgcagac 540 aaaactcaca catgcccacc gtgcccagca cctgaactcc tggggggacc gtcagtcttc 600 ctcttccccc caaaacccaa ggacaccctc atgatctccc ggacccctga ggtcacatgc 660 gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt tcaactggta cgtggacggc 720 gtggaggtgc ataatgccaa gacaaagccg cgggaggagc agtacaacag cacgtaccgt 780 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 840 aaggtctcca acaaagccct cccagccccc atcgagaaaa ccatctccaa agccaaaggg 900 cagccccgag aaccacaggt gtacaccctg cccccatgcc gggatgagct gaccaagaac 960 caggtcagcc tgtggtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1020 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1080 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1140 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1200 tccctgtctc cgggtaaatc cggaggcctg aacgacatct tcgaggccca gaagattgaa 1260 tggcacgag 1269 <![CDATA[<210> 245]]> <![CDATA[<211> 422]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 人類4-1BB抗原Fc杵鏈]]> <![CDATA[<400> 245]]> Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp 85 90 95 Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro 100 105 110 Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr 115 120 125 Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro 130 135 140 Gly Ala Ser Ser Val Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His 145 150 155 160 Ser Pro Gln Val Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Pro Lys 165 170 175 Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln 405 410 415 Lys Ile Glu Trp His Glu 420 <![CDATA[<210> 246]]> <![CDATA[<211> 422]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 獼猴4-1BB抗原]]> Fc杵鏈 <![CDATA[<400> 246]]> Leu Gln Asp Leu Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Ser Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Lys Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Ile Ser Gly Tyr His Cys Leu Gly Ala Glu Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Asp 85 90 95 Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro 100 105 110 Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr 115 120 125 Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro 130 135 140 Gly Ala Ser Ser Ala Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His 145 150 155 160 Ser Pro Gln Val Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Pro Lys 165 170 175 Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu 180 185 190 Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr 195 200 205 Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val 210 215 220 Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val 225 230 235 240 Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser 245 250 255 Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu 260 265 270 Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala 275 280 285 Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro 290 295 300 Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln 305 310 315 320 Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala 325 330 335 Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr 340 345 350 Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu 355 360 365 Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser 370 375 380 Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser 385 390 395 400 Leu Ser Pro Gly Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln 405 410 415 Lys Ile Glu Trp His Glu 420 <![CDATA[<210> 247]]> <![CDATA[<211> 423]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類4-1BB抗原Fc杵鏈]]> <![CDATA[<400> 247]]> Val Gln Asn Ser Cys Asp Asn Cys Gln Pro Gly Thr Phe Cys Arg Lys 1 5 10 15 Tyr Asn Pro Val Cys Lys Ser Cys Pro Pro Ser Thr Phe Ser Ser Ile 20 25 30 Gly Gly Gln Pro Asn Cys Asn Ile Cys Arg Val Cys Ala Gly Tyr Phe 35 40 45 Arg Phe Lys Lys Phe Cys Ser Ser Thr His Asn Ala Glu Cys Glu Cys 50 55 60 Ile Glu Gly Phe His Cys Leu Gly Pro Gln Cys Thr Arg Cys Glu Lys 65 70 75 80 Asp Cys Arg Pro Gly Gln Glu Leu Thr Lys Gln Gly Cys Lys Thr Cys 85 90 95 Ser Leu Gly Thr Phe Asn Asp Gln Asn Gly Thr Gly Val Cys Arg Pro 100 105 110 Trp Thr Asn Cys Ser Leu Asp Gly Arg Ser Val Leu Lys Thr Gly Thr 115 120 125 Thr Glu Lys Asp Val Val Cys Gly Pro Pro Val Val Ser Phe Ser Pro 130 135 140 Ser Thr Thr Ile Ser Val Thr Pro Glu Gly Gly Pro Gly Gly His Ser 145 150 155 160 Leu Gln Val Leu Val Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Pro 165 170 175 Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu 180 185 190 Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp 195 200 205 Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp 210 215 220 Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly 225 230 235 240 Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn 245 250 255 Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp 260 265 270 Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro 275 280 285 Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu 290 295 300 Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn 305 310 315 320 Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile 325 330 335 Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr 340 345 350 Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys 355 360 365 Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys 370 375 380 Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu 385 390 395 400 Ser Leu Ser Pro Gly Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala 405 410 415 Gln Lys Ile Glu Trp His Glu 420 <![CDATA[<210> 248]]> <![CDATA[<211> 28]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 引子 MS63]]> <![CDATA[<400> 248]]> tttcgcacag taatatacgg ccgtgtcc 28 <![CDATA[<210> 249]]> <![CDATA[<211> 321]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(12B3) VL]]> <![CDATA[<400> 249]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatcattcgt atccgcagac gtttggccag 300 ggcaccaaag tcgagatcaa g 321 <![CDATA[<210> 250]]> <![CDATA[<211> 360]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(12B3) VH]]> <![CDATA[<400> 250]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 <![CDATA[<210> 251]]> <![CDATA[<211> 318]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(25G7) VL]]> <![CDATA[<400> 251]]> tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60 acatgccaag gagacagcct cagaagttat tatgcaagct ggtaccagca gaagccagga 120 caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga 180 ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa 240 gatgaggctg actattactg taactccctt gataggcgcg gtatgtgggt attcggcgga 300 gggaccaagc tgaccgtc 318 <![CDATA[<210> 252]]> <![CDATA[<211> 354]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(25G7) VH]]> <![CDATA[<400> 252]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgacgac 300 ccgtggccgc cgttcgacta ctggggccaa ggaaccctgg tcaccgtctc gagt 354 <![CDATA[<210> 253]]> <![CDATA[<211> 321]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(11D5) VL]]> <![CDATA[<400> 253]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag cttaattcgt atcctcagac gtttggccag 300 ggcaccaaag tcgagatcaa g 321 <![CDATA[<210> 254]]> <![CDATA[<211> 357]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(11D5) VH]]> <![CDATA[<400> 254]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctact 300 ctgatctacg gttacttcga ctactggggc caagggacca ccgtgaccgt ctcctca 357 <![CDATA[<210> 255]]> <![CDATA[<211> 321]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(9B11) VL]]> <![CDATA[<400> 255]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag gttaattctt atccgcagac gtttggccag 300 ggcaccaaag tcgagatcaa g 321 <![CDATA[<210> 256]]> <![CDATA[<211> 360]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(9B11) VH]]> <![CDATA[<400> 256]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 <![CDATA[<210> 257]]> <![CDATA[<211> 318]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(20G2) VL]]> <![CDATA[<400> 257]]> gacatccaga tgacccagtc tccatccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag cagcactcgt attatacgtt tggccagggc 300 accaaagtcg agatcaag 318 <![CDATA[<210> 258]]> <![CDATA[<211> 360]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 4-1BB(20G2) VH]]> <![CDATA[<400> 258]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttac 300 tactgggaat cttacccgtt cgactactgg ggccaaggga ccaccgtgac cgtctccagc 360 <![CDATA[<210> 259]]> <![CDATA[<211> 642]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 12B3 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 259]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag tatcattcgt atccgcagac gtttggccag 300 ggcaccaaag tcgagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <![CDATA[<210> 260]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 12B3 P329GLALA IgG1重鏈]]> <![CDATA[<400> 260]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 261]]> <![CDATA[<211> 214]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 12B3 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 261]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr His Ser Tyr Pro Gln 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <![CDATA[<210> 262]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 12B3 P329GLALA IgG1重鏈]]> <![CDATA[<400> 262]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 263]]> <![CDATA[<211> 639]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 25G7 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 263]]> tcgtctgagc tgactcagga ccctgctgtg tctgtggcct tgggacagac agtcaggatc 60 acatgccaag gagacagcct cagaagttat tatgcaagct ggtaccagca gaagccagga 120 caggcccctg tacttgtcat ctatggtaaa aacaaccggc cctcagggat cccagaccga 180 ttctctggct ccagctcagg aaacacagct tccttgacca tcactggggc tcaggcggaa 240 gatgaggctg actattactg taactccctt gataggcgcg gtatgtgggt attcggcgga 300 gggaccaagc tgaccgtcct aggtcaaccc aaggctgccc ccagcgtgac cctgttcccc 360 cccagcagcg aggaactgca ggccaacaag gccaccctgg tctgcctgat cagcgacttc 420 tacccaggcg ccgtgaccgt ggcctggaag gccgacagca gccccgtgaa ggccggcgtg 480 gagaccacca cccccagcaa gcagagcaac aacaagtacg ccgccagcag ctacctgagc 540 ctgacccccg agcagtggaa gagccacagg tcctacagct gccaggtgac ccacgagggc 600 agcaccgtgg agaaaaccgt ggcccccacc gagtgcagc 639 <![CDATA[<210> 264]]> <![CDATA[<211> 1344]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 25G7 P329GLALA IgG1重鏈]]> <![CDATA[<400> 264]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgacgac 300 ccgtggccgc cgttcgacta ctggggccaa ggaaccctgg tcaccgtctc gagtgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctgac ctgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <![CDATA[<210> 265]]> <![CDATA[<211> 213]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 25G7 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 265]]> Ser Ser Glu Leu Thr Gln Asp Pro Ala Val Ser Val Ala Leu Gly Gln 1 5 10 15 Thr Val Arg Ile Thr Cys Gln Gly Asp Ser Leu Arg Ser Tyr Tyr Ala 20 25 30 Ser Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Val Leu Val Ile Tyr 35 40 45 Gly Lys Asn Asn Arg Pro Ser Gly Ile Pro Asp Arg Phe Ser Gly Ser 50 55 60 Ser Ser Gly Asn Thr Ala Ser Leu Thr Ile Thr Gly Ala Gln Ala Glu 65 70 75 80 Asp Glu Ala Asp Tyr Tyr Cys Asn Ser Leu Asp Arg Arg Gly Met Trp 85 90 95 Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gln Pro Lys Ala 100 105 110 Ala Pro Ser Val Thr Leu Phe Pro Pro Ser Ser Glu Glu Leu Gln Ala 115 120 125 Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asp Phe Tyr Pro Gly Ala 130 135 140 Val Thr Val Ala Trp Lys Ala Asp Ser Ser Pro Val Lys Ala Gly Val 145 150 155 160 Glu Thr Thr Thr Pro Ser Lys Gln Ser Asn Asn Lys Tyr Ala Ala Ser 165 170 175 Ser Tyr Leu Ser Leu Thr Pro Glu Gln Trp Lys Ser His Arg Ser Tyr 180 185 190 Ser Cys Gln Val Thr His Glu Gly Ser Thr Val Glu Lys Thr Val Ala 195 200 205 Pro Thr Glu Cys Ser 210 <![CDATA[<210> 266]]> <![CDATA[<211> 448]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 25G7 P329GLALA IgG1重鏈]]> <![CDATA[<400> 266]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 267]]> <![CDATA[<211> 642]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 11D5 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 267]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag cttaattcgt atcctcagac gtttggccag 300 ggcaccaaag tcgagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <![CDATA[<210> 268]]> <![CDATA[<211> 1347]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 11D5 P329GLALA IgG1重鏈]]> <![CDATA[<400> 268]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctact 300 ctgatctacg gttacttcga ctactggggc caagggacca ccgtgaccgt ctcctcagct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaagctgc agggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ggcgccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatcccg ggatgagctg 1080 accaagaacc aggtcagcct gacctgcctg gtcaaaggct tctatcccag cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200 gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 1260 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 1320 aagagcctct ccctgtctcc gggtaaa 1347 <![CDATA[<210> 269]]> <![CDATA[<211> 214]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 11D5 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 269]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Leu Asn Ser Tyr Pro Gln 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <![CDATA[<210> 270]]> <![CDATA[<211> 449]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 11D5 P329GLALA IgG1重鏈]]> <![CDATA[<400> 270]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <![CDATA[<210> 271]]> <![CDATA[<211> 642]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 9B11 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 271]]> gacatccaga tgacccagtc tccttccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag gttaattctt atccgcagac gtttggccag 300 ggcaccaaag tcgagatcaa gcgtacggtg gctgcaccat ctgtcttcat cttcccgcca 360 tctgatgagc agttgaaatc tggaactgcc tctgttgtgt gcctgctgaa taacttctat 420 cccagagagg ccaaagtaca gtggaaggtg gataacgccc tccaatcggg taactcccag 480 gagagtgtca cagagcagga cagcaaggac agcacctaca gcctcagcag caccctgacg 540 ctgagcaaag cagactacga gaaacacaaa gtctacgcct gcgaagtcac ccatcagggc 600 ctgagctcgc ccgtcacaaa gagcttcaac aggggagagt gt 642 <![CDATA[<210> 272]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 9B11 P329GLALA IgG1重鏈]]> <![CDATA[<400> 272]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 273]]> <![CDATA[<211> 214]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 9B11 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 273]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Val Asn Ser Tyr Pro Gln 85 90 95 Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala 100 105 110 Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly 115 120 125 Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 130 135 140 Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln 145 150 155 160 Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 165 170 175 Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 180 185 190 Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser 195 200 205 Phe Asn Arg Gly Glu Cys 210 <![CDATA[<210> 274]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 9B11 P329GLALA IgG1重鏈]]> <![CDATA[<400> 274]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 275]]> <![CDATA[<211> 639]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 20G2 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 275]]> gacatccaga tgacccagtc tccatccacc ctgtctgcat ctgtaggaga ccgtgtcacc 60 atcacttgcc gtgccagtca gagtattagt agctggttgg cctggtatca gcagaaacca 120 gggaaagccc ctaagctcct gatctatgat gcctccagtt tggaaagtgg ggtcccatca 180 cgtttcagcg gcagtggatc cgggacagaa ttcactctca ccatcagcag cttgcagcct 240 gatgattttg caacttatta ctgccaacag cagcactcgt attatacgtt tggccagggc 300 accaaagtcg agatcaagcg tacggtggct gcaccatctg tcttcatctt cccgccatct 360 gatgagcagt tgaaatctgg aactgcctct gttgtgtgcc tgctgaataa cttctatccc 420 agagaggcca aagtacagtg gaaggtggat aacgccctcc aatcgggtaa ctcccaggag 480 agtgtcacag agcaggacag caaggacagc acctacagcc tcagcagcac cctgacgctg 540 agcaaagcag actacgagaa acacaaagtc tacgcctgcg aagtcaccca tcagggcctg 600 agctcgcccg tcacaaagag cttcaacagg ggagagtgt 639 <![CDATA[<210> 276]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA 20G2 P329GLALA IgG1重鏈]]> <![CDATA[<400> 276]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttac 300 tactgggaat cttacccgtt cgactactgg ggccaaggga ccaccgtgac cgtctccagc 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 277]]> <![CDATA[<211> 213]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 20G2 P329GLALA IgG1輕鏈]]> <![CDATA[<400> 277]]> Asp Ile Gln Met Thr Gln Ser Pro Ser Thr Leu Ser Ala Ser Val Gly 1 5 10 15 Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Ser Ile Ser Ser Trp 20 25 30 Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 35 40 45 Tyr Asp Ala Ser Ser Leu Glu Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60 Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 65 70 75 80 Asp Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Gln His Ser Tyr Tyr Thr 85 90 95 Phe Gly Gln Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro 100 105 110 Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr 115 120 125 Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys 130 135 140 Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu 145 150 155 160 Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser 165 170 175 Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala 180 185 190 Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe 195 200 205 Asn Arg Gly Glu Cys 210 <![CDATA[<210> 278]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 20G2 P329GLALA IgG1重鏈]]> <![CDATA[<400> 278]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Tyr Tyr Trp Glu Ser Tyr Pro Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 279]]> <![CDATA[<211> 421]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> mu4-1BB D1 / hu4-1BB D2 Fc杵]]> <![CDATA[<400> 279]]> Val Gln Asn Ser Cys Asp Asn Cys Gln Pro Gly Thr Phe Cys Arg Lys 1 5 10 15 Tyr Asn Pro Val Cys Lys Ser Cys Pro Pro Ser Thr Phe Ser Ser Ile 20 25 30 Gly Gly Gln Pro Asn Cys Asn Ile Cys Arg Val Cys Ala Gly Tyr Phe 35 40 45 Arg Phe Lys Lys Phe Cys Ser Ser Thr His Asn Ala Glu Cys Glu Cys 50 55 60 Ile Glu Gly Phe His Cys Leu Gly Pro Gln Cys Thr Arg Cys Glu Lys 65 70 75 80 Asp Cys Arg Pro Gly Gln Glu Leu Thr Lys Gln Gly Cys Lys Asp Cys 85 90 95 Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg Pro Trp 100 105 110 Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly Thr Lys 115 120 125 Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser Pro Gly 130 135 140 Ala Ser Ser Val Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly His Ser 145 150 155 160 Pro Gln Val Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Pro Lys Ser 165 170 175 Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu 180 185 190 Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu 195 200 205 Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser 210 215 220 His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu 225 230 235 240 Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr 245 250 255 Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn 260 265 270 Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro 275 280 285 Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln 290 295 300 Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val 305 310 315 320 Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val 325 330 335 Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro 340 345 350 Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr 355 360 365 Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val 370 375 380 Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu 385 390 395 400 Ser Pro Gly Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys 405 410 415 Ile Glu Trp His Glu 420 <![CDATA[<210> 280]]> <![CDATA[<211> 424]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> hu4-1BB D1 / mu4-1BB D2 Fc杵]]> <![CDATA[<400> 280]]> Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Thr 85 90 95 Cys Ser Leu Gly Thr Phe Asn Asp Gln Asn Gly Thr Gly Val Cys Arg 100 105 110 Pro Trp Thr Asn Cys Ser Leu Asp Gly Arg Ser Val Leu Lys Thr Gly 115 120 125 Thr Thr Glu Lys Asp Val Val Cys Gly Pro Pro Val Val Ser Phe Ser 130 135 140 Pro Ser Thr Thr Ile Ser Val Thr Pro Glu Gly Gly Pro Gly Gly His 145 150 155 160 Ser Leu Gln Val Leu Val Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser 165 170 175 Pro Lys Ser Ala Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 180 185 190 Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 195 200 205 Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 210 215 220 Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 225 230 235 240 Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 245 250 255 Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 260 265 270 Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 275 280 285 Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 290 295 300 Glu Pro Gln Val Tyr Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys 305 310 315 320 Asn Gln Val Ser Leu Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 325 330 335 Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 340 345 350 Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 355 360 365 Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 370 375 380 Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 385 390 395 400 Leu Ser Leu Ser Pro Gly Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu 405 410 415 Ala Gln Lys Ile Glu Trp His Glu 420 <![CDATA[<210> 281]]> <![CDATA[<211> 354]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> hu 4-1BB D1 Fc杵]]> <![CDATA[<400> 281]]> Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys Val 85 90 95 Asp Glu Gln Leu Tyr Phe Gln Gly Gly Ser Pro Lys Ser Ala Asp Lys 100 105 110 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro 115 120 125 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 130 135 140 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 145 150 155 160 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 165 170 175 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 180 185 190 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 195 200 205 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys 210 215 220 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 225 230 235 240 Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp 245 250 255 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 260 265 270 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 275 280 285 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 290 295 300 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 305 310 315 320 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 325 330 335 Lys Ser Gly Gly Leu Asn Asp Ile Phe Glu Ala Gln Lys Ile Glu Trp 340 345 350 His Glu <![CDATA[<210> 282]]> <![CDATA[<211> 94]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類4-1BB結構域D1 (N-端)]]> <![CDATA[<400> 282]]> Val Gln Asn Ser Cys Asp Asn Cys Gln Pro Gly Thr Phe Cys Arg Lys 1 5 10 15 Tyr Asn Pro Val Cys Lys Ser Cys Pro Pro Ser Thr Phe Ser Ser Ile 20 25 30 Gly Gly Gln Pro Asn Cys Asn Ile Cys Arg Val Cys Ala Gly Tyr Phe 35 40 45 Arg Phe Lys Lys Phe Cys Ser Ser Thr His Asn Ala Glu Cys Glu Cys 50 55 60 Ile Glu Gly Phe His Cys Leu Gly Pro Gln Cys Thr Arg Cys Glu Lys 65 70 75 80 Asp Cys Arg Pro Gly Gln Glu Leu Thr Lys Gln Gly Cys Lys 85 90 <![CDATA[<210> 283]]> <![CDATA[<211> 68]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 人類4-1BB結構域D2 (C-端)]]> <![CDATA[<400> 283]]> Asp Cys Cys Phe Gly Thr Phe Asn Asp Gln Lys Arg Gly Ile Cys Arg 1 5 10 15 Pro Trp Thr Asn Cys Ser Leu Asp Gly Lys Ser Val Leu Val Asn Gly 20 25 30 Thr Lys Glu Arg Asp Val Val Cys Gly Pro Ser Pro Ala Asp Leu Ser 35 40 45 Pro Gly Ala Ser Ser Val Thr Pro Pro Ala Pro Ala Arg Glu Pro Gly 50 55 60 His Ser Pro Gln 65 <![CDATA[<210> 284]]> <![CDATA[<211> 95]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 人類4-1BB結構域D1 (N-端)]]> <![CDATA[<400> 284]]> Leu Gln Asp Pro Cys Ser Asn Cys Pro Ala Gly Thr Phe Cys Asp Asn 1 5 10 15 Asn Arg Asn Gln Ile Cys Ser Pro Cys Pro Pro Asn Ser Phe Ser Ser 20 25 30 Ala Gly Gly Gln Arg Thr Cys Asp Ile Cys Arg Gln Cys Lys Gly Val 35 40 45 Phe Arg Thr Arg Lys Glu Cys Ser Ser Thr Ser Asn Ala Glu Cys Asp 50 55 60 Cys Thr Pro Gly Phe His Cys Leu Gly Ala Gly Cys Ser Met Cys Glu 65 70 75 80 Gln Asp Cys Lys Gln Gly Gln Glu Leu Thr Lys Lys Gly Cys Lys 85 90 95 <![CDATA[<210> 285]]> <![CDATA[<211> 70]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> 鼠類4-1BB結構域D2 (C-端)]]> <![CDATA[<400> 285]]> Thr Cys Ser Leu Gly Thr Phe Asn Asp Gln Asn Gly Thr Gly Val Cys 1 5 10 15 Arg Pro Trp Thr Asn Cys Ser Leu Asp Gly Arg Ser Val Leu Lys Thr 20 25 30 Gly Thr Thr Glu Lys Asp Val Val Cys Gly Pro Pro Val Val Ser Phe 35 40 45 Ser Pro Ser Thr Thr Ile Ser Val Thr Pro Glu Gly Gly Pro Gly Gly 50 55 60 His Ser Leu Gln Val Leu 65 70 <![CDATA[<210> 286]]> <![CDATA[<211> 2076]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (12B3) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 286]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc cgtgttccct ctggcccctt ccagcaagtc tacctctggc 420 ggcacagccg ctctgggctg cctcgtgaag gactacttcc ccgagcctgt gacagtgtcc 480 tggaactctg gcgccctgac atccggcgtg cacacctttc cagctgtgct gcagtcctcc 540 ggcctgtact ccctgtcctc cgtcgtgaca gtgccctcca gctctctggg cacccagacc 600 tacatctgca acgtgaacca caagccctcc aacaccaagg tggacaagaa ggtggaaccc 660 aagtcctgcg acaagaccca cacctgtccc ccttgtcctg cccctgaagc tgctggcggc 720 cctagcgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatctc ccggaccccc 780 gaagtgacct gcgtggtggt ggatgtgtcc cacgaggacc ctgaagtgaa gttcaattgg 840 tacgtggacg gcgtggaagt gcacaatgcc aagaccaagc ctagagagga acagtacaac 900 tccacctacc gggtggtgtc cgtgctgacc gtgctgcacc aggattggct gaacggcaaa 960 gagtacaagt gcaaggtgtc caacaaggcc ctgggagccc ccatcgaaaa gaccatctcc 1020 aaggccaagg gccagcctcg cgagcctcag gtgtacaccc tgccccctag cagagatgag 1080 ctgaccaaga accaggtgtc cctgacctgt ctcgtgaaag gcttctaccc ctccgatatc 1140 gccgtggaat gggagagcaa cggccagccc gagaacaact acaagaccac cccccctgtg 1200 ctggactccg acggctcatt cttcctgtac tctaagctga cagtggacaa gtcccggtgg 1260 cagcagggca acgtgttctc ctgctccgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagtccc tgtccctgtc tcccggggga ggcggaggat ctggcggagg cggatccggt 1380 ggtggcggat ctgggggcgg tggatctgag gtgcagctgc tggaatctgg gggaggactg 1440 gtgcagccag gcggatctct gaggctgtcc tgcgctgctt ccggctttac cttctccagc 1500 cacgccatga gttgggtgcg ccaggcaccc ggaaaaggac tggaatgggt gtcagccatc 1560 tgggcctccg gcgagcagta ctacgccgat agcgtgaagg gccggttcac catctctcgg 1620 gataacagca agaatactct gtacctgcag atgaactccc tgcgcgctga agataccgct 1680 gtgtattact gcgccaaggg ctggctgggc aacttcgatt actggggcca gggaaccctc 1740 gtgactgtct cgagcgcttc tgtggccgct ccctccgtgt tcatcttccc accttccgac 1800 gagcagctga agtccggcac tgcctctgtc gtgtgcctgc tgaacaactt ctaccctcgg 1860 gaagccaagg tgcagtggaa agtggataac gccctgcagt ccggcaactc ccaggaatcc 1920 gtgaccgagc aggactccaa ggacagcacc tactccctga gcagcaccct gaccctgtcc 1980 aaggccgact acgagaagca caaggtgtac gcctgtgaag tgacccacca gggcctgtcc 2040 agccccgtga ccaagtcctt caaccggggc gagtgc 2076 <![CDATA[<210> 287]]> <![CDATA[<211> 692]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (12B3) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 287]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu 465 470 475 480 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 485 490 495 Thr Phe Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys 500 505 510 Gly Leu Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr 515 520 525 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 530 535 540 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 545 550 555 560 Val Tyr Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly 565 570 575 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser 580 585 590 Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala 595 600 605 Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 610 615 620 Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 625 630 635 640 Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr 645 650 655 Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys 660 665 670 Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn 675 680 685 Arg Gly Glu Cys 690 <![CDATA[<210> 288]]> <![CDATA[<211> 2070]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (25G7) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 288]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgacgac 300 ccgtggccgc cgttcgacta ctggggccaa ggaaccctgg tcaccgtctc gagtgctagc 360 accaagggcc catccgtgtt ccctctggcc ccttccagca agtctacctc tggcggcaca 420 gccgctctgg gctgcctcgt gaaggactac ttccccgagc ctgtgacagt gtcctggaac 480 tctggcgccc tgacatccgg cgtgcacacc tttccagctg tgctgcagtc ctccggcctg 540 tactccctgt cctccgtcgt gacagtgccc tccagctctc tgggcaccca gacctacatc 600 tgcaacgtga accacaagcc ctccaacacc aaggtggaca agaaggtgga acccaagtcc 660 tgcgacaaga cccacacctg tcccccttgt cctgcccctg aagctgctgg cggccctagc 720 gtgttcctgt tccccccaaa gcccaaggac accctgatga tctcccggac ccccgaagtg 780 acctgcgtgg tggtggatgt gtcccacgag gaccctgaag tgaagttcaa ttggtacgtg 840 gacggcgtgg aagtgcacaa tgccaagacc aagcctagag aggaacagta caactccacc 900 taccgggtgg tgtccgtgct gaccgtgctg caccaggatt ggctgaacgg caaagagtac 960 aagtgcaagg tgtccaacaa ggccctggga gcccccatcg aaaagaccat ctccaaggcc 1020 aagggccagc ctcgcgagcc tcaggtgtac accctgcccc ctagcagaga tgagctgacc 1080 aagaaccagg tgtccctgac ctgtctcgtg aaaggcttct acccctccga tatcgccgtg 1140 gaatgggaga gcaacggcca gcccgagaac aactacaaga ccaccccccc tgtgctggac 1200 tccgacggct cattcttcct gtactctaag ctgacagtgg acaagtcccg gtggcagcag 1260 ggcaacgtgt tctcctgctc cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgtccc tgtctcccgg gggaggcgga ggatctggcg gaggcggatc cggtggtggc 1380 ggatctgggg gcggtggatc tgaggtgcag ctgctggaat ctgggggagg actggtgcag 1440 ccaggcggat ctctgaggct gtcctgcgct gcttccggct ttaccttctc cagccacgcc 1500 atgagttggg tgcgccaggc acccggaaaa ggactggaat gggtgtcagc catctgggcc 1560 tccggcgagc agtactacgc cgatagcgtg aagggccggt tcaccatctc tcgggataac 1620 agcaagaata ctctgtacct gcagatgaac tccctgcgcg ctgaagatac cgctgtgtat 1680 tactgcgcca agggctggct gggcaacttc gattactggg gccagggaac cctcgtgact 1740 gtctcgagcg cttctgtggc cgctccctcc gtgttcatct tcccaccttc cgacgagcag 1800 ctgaagtccg gcactgcctc tgtcgtgtgc ctgctgaaca acttctaccc tcgggaagcc 1860 aaggtgcagt ggaaagtgga taacgccctg cagtccggca actcccagga atccgtgacc 1920 gagcaggact ccaaggacag cacctactcc ctgagcagca ccctgaccct gtccaaggcc 1980 gactacgaga agcacaaggt gtacgcctgt gaagtgaccc accagggcct gtccagcccc 2040 gtgaccaagt ccttcaaccg gggcgagtgc 2070 <![CDATA[<210> 289]]> <![CDATA[<211> 690]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (25G7) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 289]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp 515 520 525 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 530 535 540 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 545 550 555 560 Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly 565 570 575 Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val Phe 580 585 590 Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser Val 595 600 605 Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln Trp 610 615 620 Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val Thr 625 630 635 640 Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu Thr 645 650 655 Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu Val 660 665 670 Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg Gly 675 680 685 Glu Cys 690 <![CDATA[<210> 290]]> <![CDATA[<211> 2073]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (11D5) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 290]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctact 300 ctgatctacg gttacttcga ctactggggc caagggacca ccgtgaccgt ctcctcagct 360 agcaccaagg gcccatccgt gttccctctg gccccttcca gcaagtctac ctctggcggc 420 acagccgctc tgggctgcct cgtgaaggac tacttccccg agcctgtgac agtgtcctgg 480 aactctggcg ccctgacatc cggcgtgcac acctttccag ctgtgctgca gtcctccggc 540 ctgtactccc tgtcctccgt cgtgacagtg ccctccagct ctctgggcac ccagacctac 600 atctgcaacg tgaaccacaa gccctccaac accaaggtgg acaagaaggt ggaacccaag 660 tcctgcgaca agacccacac ctgtccccct tgtcctgccc ctgaagctgc tggcggccct 720 agcgtgttcc tgttcccccc aaagcccaag gacaccctga tgatctcccg gacccccgaa 780 gtgacctgcg tggtggtgga tgtgtcccac gaggaccctg aagtgaagtt caattggtac 840 gtggacggcg tggaagtgca caatgccaag accaagccta gagaggaaca gtacaactcc 900 acctaccggg tggtgtccgt gctgaccgtg ctgcaccagg attggctgaa cggcaaagag 960 tacaagtgca aggtgtccaa caaggccctg ggagccccca tcgaaaagac catctccaag 1020 gccaagggcc agcctcgcga gcctcaggtg tacaccctgc cccctagcag agatgagctg 1080 accaagaacc aggtgtccct gacctgtctc gtgaaaggct tctacccctc cgatatcgcc 1140 gtggaatggg agagcaacgg ccagcccgag aacaactaca agaccacccc ccctgtgctg 1200 gactccgacg gctcattctt cctgtactct aagctgacag tggacaagtc ccggtggcag 1260 cagggcaacg tgttctcctg ctccgtgatg cacgaggccc tgcacaacca ctacacccag 1320 aagtccctgt ccctgtctcc cgggggaggc ggaggatctg gcggaggcgg atccggtggt 1380 ggcggatctg ggggcggtgg atctgaggtg cagctgctgg aatctggggg aggactggtg 1440 cagccaggcg gatctctgag gctgtcctgc gctgcttccg gctttacctt ctccagccac 1500 gccatgagtt gggtgcgcca ggcacccgga aaaggactgg aatgggtgtc agccatctgg 1560 gcctccggcg agcagtacta cgccgatagc gtgaagggcc ggttcaccat ctctcgggat 1620 aacagcaaga atactctgta cctgcagatg aactccctgc gcgctgaaga taccgctgtg 1680 tattactgcg ccaagggctg gctgggcaac ttcgattact ggggccaggg aaccctcgtg 1740 actgtctcga gcgcttctgt ggccgctccc tccgtgttca tcttcccacc ttccgacgag 1800 cagctgaagt ccggcactgc ctctgtcgtg tgcctgctga acaacttcta ccctcgggaa 1860 gccaaggtgc agtggaaagt ggataacgcc ctgcagtccg gcaactccca ggaatccgtg 1920 accgagcagg actccaagga cagcacctac tccctgagca gcaccctgac cctgtccaag 1980 gccgactacg agaagcacaa ggtgtacgcc tgtgaagtga cccaccaggg cctgtccagc 2040 cccgtgacca agtccttcaa ccggggcgag tgc 2073 <![CDATA[<210> 291]]> <![CDATA[<211> 691]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (11D5) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 291]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Thr 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 450 455 460 Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val 465 470 475 480 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 485 490 495 Phe Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 500 505 510 Leu Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala 515 520 525 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 530 535 540 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 545 550 555 560 Tyr Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln 565 570 575 Gly Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val 580 585 590 Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala Ser 595 600 605 Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gln 610 615 620 Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser Val 625 630 635 640 Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu 645 650 655 Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu 660 665 670 Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg 675 680 685 Gly Glu Cys 690 <![CDATA[<210> 292]]> <![CDATA[<211> 2076]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (9B11) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 292]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc cgtgttccct ctggcccctt ccagcaagtc tacctctggc 420 ggcacagccg ctctgggctg cctcgtgaag gactacttcc ccgagcctgt gacagtgtcc 480 tggaactctg gcgccctgac atccggcgtg cacacctttc cagctgtgct gcagtcctcc 540 ggcctgtact ccctgtcctc cgtcgtgaca gtgccctcca gctctctggg cacccagacc 600 tacatctgca acgtgaacca caagccctcc aacaccaagg tggacaagaa ggtggaaccc 660 aagtcctgcg acaagaccca cacctgtccc ccttgtcctg cccctgaagc tgctggcggc 720 cctagcgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatctc ccggaccccc 780 gaagtgacct gcgtggtggt ggatgtgtcc cacgaggacc ctgaagtgaa gttcaattgg 840 tacgtggacg gcgtggaagt gcacaatgcc aagaccaagc ctagagagga acagtacaac 900 tccacctacc gggtggtgtc cgtgctgacc gtgctgcacc aggattggct gaacggcaaa 960 gagtacaagt gcaaggtgtc caacaaggcc ctgggagccc ccatcgaaaa gaccatctcc 1020 aaggccaagg gccagcctcg cgagcctcag gtgtacaccc tgccccctag cagagatgag 1080 ctgaccaaga accaggtgtc cctgacctgt ctcgtgaaag gcttctaccc ctccgatatc 1140 gccgtggaat gggagagcaa cggccagccc gagaacaact acaagaccac cccccctgtg 1200 ctggactccg acggctcatt cttcctgtac tctaagctga cagtggacaa gtcccggtgg 1260 cagcagggca acgtgttctc ctgctccgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagtccc tgtccctgtc tcccggggga ggcggaggat ctggcggagg cggatccggt 1380 ggtggcggat ctgggggcgg tggatctgag gtgcagctgc tggaatctgg gggaggactg 1440 gtgcagccag gcggatctct gaggctgtcc tgcgctgctt ccggctttac cttctccagc 1500 cacgccatga gttgggtgcg ccaggcaccc ggaaaaggac tggaatgggt gtcagccatc 1560 tgggcctccg gcgagcagta ctacgccgat agcgtgaagg gccggttcac catctctcgg 1620 gataacagca agaatactct gtacctgcag atgaactccc tgcgcgctga agataccgct 1680 gtgtattact gcgccaaggg ctggctgggc aacttcgatt actggggcca gggaaccctc 1740 gtgactgtct cgagcgcttc tgtggccgct ccctccgtgt tcatcttccc accttccgac 1800 gagcagctga agtccggcac tgcctctgtc gtgtgcctgc tgaacaactt ctaccctcgg 1860 gaagccaagg tgcagtggaa agtggataac gccctgcagt ccggcaactc ccaggaatcc 1920 gtgaccgagc aggactccaa ggacagcacc tactccctga gcagcaccct gaccctgtcc 1980 aaggccgact acgagaagca caaggtgtac gcctgtgaag tgacccacca gggcctgtcc 2040 agccccgtga ccaagtcctt caaccggggc gagtgc 2076 <![CDATA[<210> 293]]> <![CDATA[<211> 692]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (9B11) VHCH1-重鏈-(28H1) VHCL]]> <![CDATA[<400> 293]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu 465 470 475 480 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 485 490 495 Thr Phe Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys 500 505 510 Gly Leu Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr 515 520 525 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 530 535 540 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 545 550 555 560 Val Tyr Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly 565 570 575 Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser 580 585 590 Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala 595 600 605 Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 610 615 620 Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 625 630 635 640 Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr 645 650 655 Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys 660 665 670 Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn 675 680 685 Arg Gly Glu Cys 690 <![CDATA[<210> 294]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (12B3) VHCH1-重鏈杵]]> <![CDATA[<400> 294]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggccctag cgtgttccct ctggccccta gcagcaagag cacaagtgga 420 ggaacagccg ccctgggctg cctggtcaag gactacttcc ccgagcccgt gaccgtgtcc 480 tggaattctg gcgccctgac aagcggcgtg cacacatttc cagccgtgct gcagagcagc 540 ggcctgtact ctctgagcag cgtcgtgacc gtgccctcta gctctctggg cacccagacc 600 tacatctgca acgtgaacca caagcccagc aacaccaaag tggacaagaa ggtggaaccc 660 aagagctgcg acaagaccca cacctgtccc ccttgccctg cccctgaagc tgctggtggc 720 ccttccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc 780 gaagtgacct gcgtggtggt cgatgtgtcc cacgaggacc ctgaagtgaa gttcaattgg 840 tacgtggacg gcgtggaagt gcacaatgcc aagaccaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatg ccgggatgag 1080 ctgaccaaga accaggtcag cctgtggtgc ctggtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 295]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (12B3) VHCH1-重鏈杵]]> <![CDATA[<400> 295]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 296]]> <![CDATA[<211> 1344]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (25G7) VHCH1-重鏈杵]]> <![CDATA[<400> 296]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgacgac 300 ccgtggccgc cgttcgacta ctggggccaa ggaaccctgg tcaccgtctc gagtgctagc 360 accaagggcc ctagcgtgtt ccctctggcc cctagcagca agagcacaag tggaggaaca 420 gccgccctgg gctgcctggt caaggactac ttccccgagc ccgtgaccgt gtcctggaat 480 tctggcgccc tgacaagcgg cgtgcacaca tttccagccg tgctgcagag cagcggcctg 540 tactctctga gcagcgtcgt gaccgtgccc tctagctctc tgggcaccca gacctacatc 600 tgcaacgtga accacaagcc cagcaacacc aaagtggaca agaaggtgga acccaagagc 660 tgcgacaaga cccacacctg tcccccttgc cctgcccctg aagctgctgg tggcccttcc 720 gtgttcctgt tccccccaaa gcccaaggac accctgatga tcagccggac ccccgaagtg 780 acctgcgtgg tggtcgatgt gtcccacgag gaccctgaag tgaagttcaa ttggtacgtg 840 gacggcgtgg aagtgcacaa tgccaagacc aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc catgccggga tgagctgacc 1080 aagaaccagg tcagcctgtg gtgcctggtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct ctacagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg taaa 1344 <![CDATA[<210> 297]]> <![CDATA[<211> 448]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (25G7) VHCH1-重鏈杵]]> <![CDATA[<400> 297]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 298]]> <![CDATA[<211> 1347]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (11D5) VHCH1-重鏈杵]]> <![CDATA[<400> 298]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctact 300 ctgatctacg gttacttcga ctactggggc caagggacca ccgtgaccgt ctcctcagct 360 agcaccaagg gccctagcgt gttccctctg gcccctagca gcaagagcac aagtggagga 420 acagccgccc tgggctgcct ggtcaaggac tacttccccg agcccgtgac cgtgtcctgg 480 aattctggcg ccctgacaag cggcgtgcac acatttccag ccgtgctgca gagcagcggc 540 ctgtactctc tgagcagcgt cgtgaccgtg ccctctagct ctctgggcac ccagacctac 600 atctgcaacg tgaaccacaa gcccagcaac accaaagtgg acaagaaggt ggaacccaag 660 agctgcgaca agacccacac ctgtccccct tgccctgccc ctgaagctgc tggtggccct 720 tccgtgttcc tgttcccccc aaagcccaag gacaccctga tgatcagccg gacccccgaa 780 gtgacctgcg tggtggtcga tgtgtcccac gaggaccctg aagtgaagtt caattggtac 840 gtggacggcg tggaagtgca caatgccaag accaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ggcgccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tacaccctgc ccccatgccg ggatgagctg 1080 accaagaacc aggtcagcct gtggtgcctg gtcaaaggct tctatcccag cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200 gactccgacg gctccttctt cctctacagc aagctcaccg tggacaagag caggtggcag 1260 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 1320 aagagcctct ccctgtctcc gggtaaa 1347 <![CDATA[<210> 299]]> <![CDATA[<211> 449]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (11D5) VHCH1-重鏈杵]]> <![CDATA[<400> 299]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Lys <![CDATA[<210> 300]]> <![CDATA[<211> 1350]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (9B11) VHCH1-重鏈杵]]> <![CDATA[<400> 300]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggccctag cgtgttccct ctggccccta gcagcaagag cacaagtgga 420 ggaacagccg ccctgggctg cctggtcaag gactacttcc ccgagcccgt gaccgtgtcc 480 tggaattctg gcgccctgac aagcggcgtg cacacatttc cagccgtgct gcagagcagc 540 ggcctgtact ctctgagcag cgtcgtgacc gtgccctcta gctctctggg cacccagacc 600 tacatctgca acgtgaacca caagcccagc aacaccaaag tggacaagaa ggtggaaccc 660 aagagctgcg acaagaccca cacctgtccc ccttgccctg cccctgaagc tgctggtggc 720 ccttccgtgt tcctgttccc cccaaagccc aaggacaccc tgatgatcag ccggaccccc 780 gaagtgacct gcgtggtggt cgatgtgtcc cacgaggacc ctgaagtgaa gttcaattgg 840 tacgtggacg gcgtggaagt gcacaatgcc aagaccaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccatg ccgggatgag 1080 ctgaccaaga accaggtcag cctgtggtgc ctggtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtaaa 1350 <![CDATA[<210> 301]]> <![CDATA[<211> 450]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (9B11) VHCH1-重鏈杵]]> <![CDATA[<400> 301]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Lys 450 <![CDATA[<210> 302]]> <![CDATA[<211> 1338]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (8H9) VHCH1-重鏈杵]]> <![CDATA[<400> 302]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 ggttggatgg actactgggg ccaagggacc accgtgaccg tctcctcagc tagcaccaag 360 ggccctagcg tgttccctct ggcccctagc agcaagagca caagtggagg aacagccgcc 420 ctgggctgcc tggtcaagga ctacttcccc gagcccgtga ccgtgtcctg gaattctggc 480 gccctgacaa gcggcgtgca cacatttcca gccgtgctgc agagcagcgg cctgtactct 540 ctgagcagcg tcgtgaccgt gccctctagc tctctgggca cccagaccta catctgcaac 600 gtgaaccaca agcccagcaa caccaaagtg gacaagaagg tggaacccaa gagctgcgac 660 aagacccaca cctgtccccc ttgccctgcc cctgaagctg ctggtggccc ttccgtgttc 720 ctgttccccc caaagcccaa ggacaccctg atgatcagcc ggacccccga agtgacctgc 780 gtggtggtcg atgtgtccca cgaggaccct gaagtgaagt tcaattggta cgtggacggc 840 gtggaagtgc acaatgccaa gaccaagccg cgggaggagc agtacaacag cacgtaccgt 900 gtggtcagcg tcctcaccgt cctgcaccag gactggctga atggcaagga gtacaagtgc 960 aaggtctcca acaaagccct cggcgccccc atcgagaaaa ccatctccaa agccaaaggg 1020 cagccccgag aaccacaggt gtacaccctg cccccatgcc gggatgagct gaccaagaac 1080 caggtcagcc tgtggtgcct ggtcaaaggc ttctatccca gcgacatcgc cgtggagtgg 1140 gagagcaatg ggcagccgga gaacaactac aagaccacgc ctcccgtgct ggactccgac 1200 ggctccttct tcctctacag caagctcacc gtggacaaga gcaggtggca gcaggggaac 1260 gtcttctcat gctccgtgat gcatgaggct ctgcacaacc actacacgca gaagagcctc 1320 tccctgtctc cgggtaaa 1338 <![CDATA[<210> 303]]> <![CDATA[<211> 446]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (8H9) VHCH1-重鏈杵]]> <![CDATA[<400> 303]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Gly Trp Met Asp Tyr Trp Gly Gln Gly Thr Thr Val 100 105 110 Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala 115 120 125 Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu 130 135 140 Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly 145 150 155 160 Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser 165 170 175 Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu 180 185 190 Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr 195 200 205 Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr 210 215 220 Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser Val Phe 225 230 235 240 Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro 245 250 255 Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val 260 265 270 Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr 275 280 285 Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val 290 295 300 Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys 305 310 315 320 Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr Ile Ser 325 330 335 Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro 340 345 350 Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys Leu Val 355 360 365 Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly 370 375 380 Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp 385 390 395 400 Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp 405 410 415 Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His 420 425 430 Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 435 440 445 <![CDATA[<210> 304]]> <![CDATA[<211> 1752]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc杵VH (4B9)之核苷酸序列 ]]> <![CDATA[<400> 304]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc cctgcagaga tgagctgacc 1080 aagaaccagg tgtccctgtg gtgtctggtc aagggcttct accccagcga tatcgccgtg 1140 gagtgggaga gcaacggcca gcctgagaac aactacaaga ccaccccccc tgtgctggac 1200 agcgacggca gcttcttcct gtactccaaa ctgaccgtgg acaagagccg gtggcagcag 1260 ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgagcc tgagccccgg cggaggcggc ggaagcggag gaggaggatc tgggggcgga 1380 ggttccggag gcggaggatc cgaggtgcag ctgctcgaaa gcggcggagg actggtgcag 1440 cctggcggca gcctgagact gtcttgcgcc gccagcggct tcaccttcag cagctacgcc 1500 atgagctggg tccgccaggc ccctggcaag ggactggaat gggtgtccgc catcatcggc 1560 tctggcgcca gcacctacta cgccgacagc gtgaagggcc ggttcaccat cagccgggac 1620 aacagcaaga acaccctgta cctgcagatg aacagcctgc gggccgagga caccgccgtg 1680 tactactgcg ccaagggatg gttcggcggc ttcaactact ggggacaggg caccctggtc 1740 accgtgtcca gc 1752 <![CDATA[<210> 305]]> <![CDATA[<211> 1725]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc臼VL (4B9)之核苷酸序列]]> <![CDATA[<400> 305]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtgc accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctctc gtgcgcagtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct cgtgagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg tggaggcggc ggaagcggag gaggaggatc cggcggcgga 1380 ggttccggag gcggtggatc tgagatcgtg ctgacccagt ctcccggcac cctgtctctg 1440 agccctggcg agagagccac cctgtcctgc agagcctccc agtccgtgac ctcctcctac 1500 ctcgcctggt atcagcagaa gcccggccag gcccctcggc tgctgatcaa cgtgggcagt 1560 cggagagcca ccggcatccc tgaccggttc tccggctctg gctccggcac cgacttcacc 1620 ctgaccatct cccggctgga acccgaggac ttcgccgtgt actactgcca gcagggcatc 1680 atgctgcccc ccacctttgg ccagggcacc aaggtggaaa tcaag 1725 <![CDATA[<210> 306]]> <![CDATA[<211> 584]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 306]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr Ala 515 520 525 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 530 535 540 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 545 550 555 560 Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gln 565 570 575 Gly Thr Leu Val Thr Val Ser Ser 580 <![CDATA[<210> 307]]> <![CDATA[<211> 575]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 307]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 465 470 475 480 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 485 490 495 Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 500 505 510 Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile Pro Asp 515 520 525 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 530 535 540 Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Ile 545 550 555 560 Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 308]]> <![CDATA[<211> 1749]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc杵VH (28H1)之核苷酸序列]]> <![CDATA[<400> 308]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc cctgcagaga tgagctgacc 1080 aagaaccagg tgtccctgtg gtgtctggtc aagggcttct accccagcga tatcgccgtg 1140 gagtgggaga gcaacggcca gcctgagaac aactacaaga ccaccccccc tgtgctggac 1200 agcgacggca gcttcttcct gtactccaaa ctgaccgtgg acaagagccg gtggcagcag 1260 ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgagcc tgagccccgg cggaggcggc ggaagcggag gaggaggatc cggaggaggg 1380 ggaagtggcg gcggaggatc tgaggtgcag ctgctggaat ccggcggagg cctggtgcag 1440 cctggcggat ctctgagact gtcctgcgcc gcctccggct tcaccttctc ctcccacgcc 1500 atgtcctggg tccgacaggc tcctggcaaa ggcctggaat gggtgtccgc catctgggcc 1560 tccggcgagc agtactacgc cgactctgtg aagggccggt tcaccatctc ccgggacaac 1620 tccaagaaca ccctgtacct gcagatgaac tccctgcggg ccgaggacac cgccgtgtac 1680 tactgtgcca agggctggct gggcaacttc gactactggg gccagggcac cctggtcacc 1740 gtgtccagc 1749 <![CDATA[<210> 309]]> <![CDATA[<211> 1725]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc臼VL (28H1)之核苷酸序列]]> <![CDATA[<400> 309]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtgc accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctctc gtgcgcagtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct cgtgagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg tggaggcggc ggaagcggag gaggaggatc cggtggtggc 1380 ggatctgggg gcggtggatc tgagatcgtg ctgacccagt ctcccggcac cctgagcctg 1440 agccctggcg agagagccac cctgagctgc agagccagcc agagcgtgag ccggagctac 1500 ctggcctggt atcagcagaa gcccggccag gcccccagac tgctgatcat cggcgccagc 1560 acccgggcca ccggcatccc cgatagattc agcggcagcg gctccggcac cgacttcacc 1620 ctgaccatca gccggctgga acccgaggac ttcgccgtgt actactgcca gcagggccag 1680 gtgatccccc ccaccttcgg ccagggcacc aaggtggaaa tcaag 1725 <![CDATA[<210> 310]]> <![CDATA[<211> 583]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc杵VH (28H1)]]> <![CDATA[<400> 310]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser His Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Trp Ala Ser Gly Glu Gln Tyr Tyr Ala Asp 515 520 525 Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr 530 535 540 Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr 545 550 555 560 Tyr Cys Ala Lys Gly Trp Leu Gly Asn Phe Asp Tyr Trp Gly Gln Gly 565 570 575 Thr Leu Val Thr Val Ser Ser 580 <![CDATA[<210> 311]]> <![CDATA[<211> 575]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc臼VL (28H1)]]> <![CDATA[<400> 311]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 465 470 475 480 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 485 490 495 Ser Arg Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 500 505 510 Arg Leu Leu Ile Ile Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Asp 515 520 525 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 530 535 540 Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Gln 545 550 555 560 Val Ile Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 312]]> <![CDATA[<211> 1746]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc杵VH (DP47)之核苷酸序列]]> <![CDATA[<400> 312]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc cctgcagaga tgagctgacc 1080 aagaaccagg tgtccctgtg gtgtctggtc aagggcttct accccagcga tatcgccgtg 1140 gagtgggaga gcaacggcca gcctgagaac aactacaaga ccaccccccc tgtgctggac 1200 agcgacggca gcttcttcct gtactccaaa ctgaccgtgg acaagagccg gtggcagcag 1260 ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgagcc tgagccccgg cggaggcggc ggaagcggag gaggaggatc cggaggaggg 1380 ggaagtggcg gcggaggatc tgaggtgcaa ttgttggagt ctgggggagg cttggtacag 1440 cctggggggt ccctgagact ctcctgtgca gccagcggat tcacctttag cagttatgcc 1500 atgagctggg tccgccaggc tccagggaag gggctggagt gggtctcagc tattagtggt 1560 agtggtggta gcacatacta cgcagactcc gtgaagggcc ggttcaccat ctccagagac 1620 aattccaaga acacgctgta tctgcagatg aacagcctga gagccgagga cacggccgta 1680 tattactgtg cgaaaggcag cggatttgac tactggggcc aaggaaccct ggtcaccgtc 1740 tcgagc 1746 <![CDATA[<210> 313]]> <![CDATA[<211> 1725]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc臼VL (DP47)之核苷酸序列]]> <![CDATA[<400> 313]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagagaatac 300 taccgtggtc cgtacgacta ctggggccaa gggaccaccg tgaccgtctc ctcagctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtgc accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctctc gtgcgcagtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct cgtgagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg tggaggcggc ggaagcggag gaggaggatc cggaggcggc 1380 ggaagcggag ggggaggctc tgaaattgtg ctgacccaga gccccggcac cctgtcactg 1440 tctccaggcg aaagagccac cctgagctgc agagccagcc agagcgtgtc cagctcttac 1500 ctggcctggt atcagcagaa gcccggacag gcccccagac tgctgatcta cggcgcctct 1560 tctagagcca ccggcatccc cgatagattc agcggcagcg gctccggcac cgacttcacc 1620 ctgacaatca gcagactgga acccgaggac tttgccgtgt attactgcca gcagtacggc 1680 agcagccccc tgacctttgg ccagggcacc aaggtggaaa tcaaa 1725 <![CDATA[<210> 314]]> <![CDATA[<211> 582]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc杵VH (DP47)]]> <![CDATA[<400> 314]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala 515 520 525 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 530 535 540 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 545 550 555 560 Tyr Tyr Cys Ala Lys Gly Ser Gly Phe Asp Tyr Trp Gly Gln Gly Thr 565 570 575 Leu Val Thr Val Ser Ser 580 <![CDATA[<210> 315]]> <![CDATA[<211> 575]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (49B4) VHCH1 Fc臼VL (DP47)]]> <![CDATA[<400> 315]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Glu Tyr Tyr Arg Gly Pro Tyr Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 465 470 475 480 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 485 490 495 Ser Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 500 505 510 Arg Leu Leu Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp 515 520 525 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 530 535 540 Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly 545 550 555 560 Ser Ser Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 316]]> <![CDATA[<211> 1758]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (12B3) VHCH1 Fc杵VH (4B9) (HC1)]]> <![CDATA[<400> 316]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctggagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcgagt 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccctg cagagatgag 1080 ctgaccaaga accaggtgtc cctgtggtgt ctggtcaagg gcttctaccc cagcgatatc 1140 gccgtggagt gggagagcaa cggccagcct gagaacaact acaagaccac cccccctgtg 1200 ctggacagcg acggcagctt cttcctgtac tccaaactga ccgtggacaa gagccggtgg 1260 cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagtccc tgagcctgag ccccggcgga ggcggcggaa gcggaggagg aggatctggg 1380 ggcggaggtt ccggaggcgg tggatctgag gtgcagctgc tcgaaagcgg cggaggactg 1440 gtgcagcctg gcggcagcct gagactgtct tgcgccgcca gcggcttcac cttcagcagc 1500 tacgccatga gctgggtccg ccaggcccct ggcaagggac tggaatgggt gtccgccatc 1560 atcggctctg gcgccagcac ctactacgcc gacagcgtga agggccggtt caccatcagc 1620 cgggacaaca gcaagaacac cctgtacctg cagatgaaca gcctgcgggc cgaggacacc 1680 gccgtgtact actgcgccaa gggatggttc ggcggcttca actactgggg acagggcacc 1740 ctggtcaccg tgtccagc 1758 <![CDATA[<210> 317]]> <![CDATA[<211> 1731]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (12B3) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 317]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctggagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcgagt 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtgcaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctctcgtgc gcagtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctcgtg agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtgga ggcggcggaa gcggaggagg aggatccggc 1380 ggcggaggtt ccggaggcgg aggatccgag atcgtgctga cccagtctcc cggcaccctg 1440 tctctgagcc ctggcgagag agccaccctg tcctgcagag cctcccagtc cgtgacctcc 1500 tcctacctcg cctggtatca gcagaagccc ggccaggccc ctcggctgct gatcaacgtg 1560 ggcagtcgga gagccaccgg catccctgac cggttctccg gctctggctc cggcaccgac 1620 ttcaccctga ccatctcccg gctggaaccc gaggacttcg ccgtgtacta ctgccagcag 1680 ggcatcatgc tgccccccac ctttggccag ggcaccaagg tggaaatcaa g 1731 <![CDATA[<210> 318]]> <![CDATA[<211> 586]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (12B3) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 318]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu 465 470 475 480 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 485 490 495 Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys 500 505 510 Gly Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr 515 520 525 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser 530 535 540 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 545 550 555 560 Ala Val Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp 565 570 575 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 580 585 <![CDATA[<210> 319]]> <![CDATA[<211> 577]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (12B3) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 319]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu 465 470 475 480 Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln 485 490 495 Ser Val Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 500 505 510 Ala Pro Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile 515 520 525 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 530 535 540 Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln 545 550 555 560 Gly Ile Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 565 570 575 Lys <![CDATA[<210> 320]]> <![CDATA[<211> 1752]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (25G7) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 320]]> gaggtgcagc tgctggaatc tggcggcgga ctggtgcagc ctggcggatc tctgagactg 60 agctgtgccg ccagcggctt caccttcagc agctacgcca tgagctgggt gcgccaggcc 120 cctggaaaag gcctggaatg ggtgtccgcc atctctggca gcggcggcag cacctactac 180 gccgattctg tgaagggccg gttcaccatc agccgggaca acagcaagaa caccctgtac 240 ctgcagatga acagcctgcg ggccgaggac accgccgtgt actattgcgc cagggacgac 300 ccctggcccc cctttgatta ttggggacag ggcaccctcg tgaccgtgtc cagcgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc cctgcagaga tgagctgacc 1080 aagaaccagg tgtccctgtg gtgtctggtc aagggcttct accccagcga tatcgccgtg 1140 gagtgggaga gcaacggcca gcctgagaac aactacaaga ccaccccccc tgtgctggac 1200 agcgacggca gcttcttcct gtactccaaa ctgaccgtgg acaagagccg gtggcagcag 1260 ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgagcc tgagccccgg cggaggcggc ggaagcggag gaggaggatc tgggggcgga 1380 ggttccggag gcggtggatc tgaggtgcag ctgctcgaaa gcggcggagg actggtgcag 1440 cctggcggca gcctgagact gtcttgcgcc gccagcggct tcaccttcag cagctacgcc 1500 atgagctggg tccgccaggc ccctggcaag ggactggaat gggtgtccgc catcatcggc 1560 tctggcgcca gcacctacta cgccgacagc gtgaagggcc ggttcaccat cagccgggac 1620 aacagcaaga acaccctgta cctgcagatg aacagcctgc gggccgagga caccgccgtg 1680 tactactgcg ccaagggatg gttcggcggc ttcaactact ggggacaggg caccctggtc 1740 accgtgtcca gc 1752 <![CDATA[<210> 321]]> <![CDATA[<211> 1725]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (25G7) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 321]]> gaggtgcagc tgctggaatc tggcggcgga ctggtgcagc ctggcggatc tctgagactg 60 agctgtgccg ccagcggctt caccttcagc agctacgcca tgagctgggt gcgccaggcc 120 cctggaaaag gcctggaatg ggtgtccgcc atctctggca gcggcggcag cacctactac 180 gccgattctg tgaagggccg gttcaccatc agccgggaca acagcaagaa caccctgtac 240 ctgcagatga acagcctgcg ggccgaggac accgccgtgt actattgcgc cagggacgac 300 ccctggcccc cctttgatta ttggggacag ggcaccctcg tgaccgtgtc cagcgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtgc accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctctc gtgcgcagtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct cgtgagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg tggaggcggc ggaagcggag gaggaggatc cggcggcgga 1380 ggttccggag gcggaggatc cgagatcgtg ctgacccagt ctcccggcac cctgtctctg 1440 agccctggcg agagagccac cctgtcctgc agagcctccc agtccgtgac ctcctcctac 1500 ctcgcctggt atcagcagaa gcccggccag gcccctcggc tgctgatcaa cgtgggcagt 1560 cggagagcca ccggcatccc tgaccggttc tccggctctg gctccggcac cgacttcacc 1620 ctgaccatct cccggctgga acccgaggac ttcgccgtgt actactgcca gcagggcatc 1680 atgctgcccc ccacctttgg ccagggcacc aaggtggaaa tcaag 1725 <![CDATA[<210> 322]]> <![CDATA[<211> 584]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (25G7) VHCH1 Fc杵VH]]> <![CDATA[<400> 322]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr Ala 515 520 525 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 530 535 540 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 545 550 555 560 Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gln 565 570 575 Gly Thr Leu Val Thr Val Ser Ser 580 <![CDATA[<210> 323]]> <![CDATA[<211> 575]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (25G7) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 323]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 465 470 475 480 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 485 490 495 Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 500 505 510 Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile Pro Asp 515 520 525 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 530 535 540 Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Ile 545 550 555 560 Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 324]]> <![CDATA[<211> 1755]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (11D5) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 324]]> caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ccggcagcag cgtgaaggtg 60 tcctgcaagg cttccggcgg caccttcagc agctacgcca tttcttgggt gcgccaggcc 120 cctggacagg gcctggaatg gatgggcggc atcatcccca tcttcggcac cgccaactac 180 gcccagaaat tccagggcag agtgaccatc accgccgaca agagcaccag caccgcctac 240 atggaactga gcagcctgcg gagcgaggac accgccgtgt actactgtgc cagaagcacc 300 ctgatctacg gctacttcga ctactggggc cagggcacca ccgtgaccgt gtctagcgct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaagctgc agggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ggcgccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tacaccctgc ccccctgcag agatgagctg 1080 accaagaacc aggtgtccct gtggtgtctg gtcaagggct tctaccccag cgatatcgcc 1140 gtggagtggg agagcaacgg ccagcctgag aacaactaca agaccacccc ccctgtgctg 1200 gacagcgacg gcagcttctt cctgtactcc aaactgaccg tggacaagag ccggtggcag 1260 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1320 aagtccctga gcctgagccc cggcggaggc ggcggaagcg gaggaggagg atctgggggc 1380 ggaggttccg gaggcggtgg atctgaggtg cagctgctcg aaagcggcgg aggactggtg 1440 cagcctggcg gcagcctgag actgtcttgc gccgccagcg gcttcacctt cagcagctac 1500 gccatgagct gggtccgcca ggcccctggc aagggactgg aatgggtgtc cgccatcatc 1560 ggctctggcg ccagcaccta ctacgccgac agcgtgaagg gccggttcac catcagccgg 1620 gacaacagca agaacaccct gtacctgcag atgaacagcc tgcgggccga ggacaccgcc 1680 gtgtactact gcgccaaggg atggttcggc ggcttcaact actggggaca gggcaccctg 1740 gtcaccgtgt ccagc 1755 <![CDATA[<210> 325]]> <![CDATA[<211> 1728]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (11D5) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 325]]> caggtgcagc tggtgcagtc tggcgccgaa gtgaagaaac ccggcagcag cgtgaaggtg 60 tcctgcaagg cttccggcgg caccttcagc agctacgcca tttcttgggt gcgccaggcc 120 cctggacagg gcctggaatg gatgggcggc atcatcccca tcttcggcac cgccaactac 180 gcccagaaat tccagggcag agtgaccatc accgccgaca agagcaccag caccgcctac 240 atggaactga gcagcctgcg gagcgaggac accgccgtgt actactgtgc cagaagcacc 300 ctgatctacg gctacttcga ctactggggc cagggcacca ccgtgaccgt gtctagcgct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaagctgc agggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ggcgccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tgcaccctgc ccccatcccg ggatgagctg 1080 accaagaacc aggtcagcct ctcgtgcgca gtcaaaggct tctatcccag cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200 gactccgacg gctccttctt cctcgtgagc aagctcaccg tggacaagag caggtggcag 1260 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 1320 aagagcctct ccctgtctcc gggtggaggc ggcggaagcg gaggaggagg atccggcggc 1380 ggaggttccg gaggcggagg atccgagatc gtgctgaccc agtctcccgg caccctgtct 1440 ctgagccctg gcgagagagc caccctgtcc tgcagagcct cccagtccgt gacctcctcc 1500 tacctcgcct ggtatcagca gaagcccggc caggcccctc ggctgctgat caacgtgggc 1560 agtcggagag ccaccggcat ccctgaccgg ttctccggct ctggctccgg caccgacttc 1620 accctgacca tctcccggct ggaacccgag gacttcgccg tgtactactg ccagcagggc 1680 atcatgctgc cccccacctt tggccagggc accaaggtgg aaatcaag 1728 <![CDATA[<210> 326]]> <![CDATA[<211> 585]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (11D5) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 326]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 450 455 460 Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val 465 470 475 480 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 485 490 495 Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 500 505 510 Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr 515 520 525 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 530 535 540 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 545 550 555 560 Val Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly 565 570 575 Gln Gly Thr Leu Val Thr Val Ser Ser 580 585 <![CDATA[<210> 327]]> <![CDATA[<211> 576]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (11D5) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 327]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser 355 360 365 Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 450 455 460 Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser 465 470 475 480 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser 485 490 495 Val Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala 500 505 510 Pro Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile Pro 515 520 525 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 530 535 540 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly 545 550 555 560 Ile Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 328]]> <![CDATA[<211> 1758]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (9B11) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 328]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccctg cagagatgag 1080 ctgaccaaga accaggtgtc cctgtggtgt ctggtcaagg gcttctaccc cagcgatatc 1140 gccgtggagt gggagagcaa cggccagcct gagaacaact acaagaccac cccccctgtg 1200 ctggacagcg acggcagctt cttcctgtac tccaaactga ccgtggacaa gagccggtgg 1260 cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagtccc tgagcctgag ccccggcgga ggcggcggaa gcggaggagg aggatctggg 1380 ggcggaggtt ccggaggcgg tggatctgag gtgcagctgc tcgaaagcgg cggaggactg 1440 gtgcagcctg gcggcagcct gagactgtct tgcgccgcca gcggcttcac cttcagcagc 1500 tacgccatga gctgggtccg ccaggcccct ggcaagggac tggaatgggt gtccgccatc 1560 atcggctctg gcgccagcac ctactacgcc gacagcgtga agggccggtt caccatcagc 1620 cgggacaaca gcaagaacac cctgtacctg cagatgaaca gcctgcgggc cgaggacacc 1680 gccgtgtact actgcgccaa gggatggttc ggcggcttca actactgggg acagggcacc 1740 ctggtcaccg tgtccagc 1758 <![CDATA[<210> 329]]> <![CDATA[<211> 1731]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (9B11) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 329]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtgcaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctctcgtgc gcagtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctcgtg agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtgga ggcggcggaa gcggaggagg aggatccggc 1380 ggcggaggtt ccggaggcgg aggatccgag atcgtgctga cccagtctcc cggcaccctg 1440 tctctgagcc ctggcgagag agccaccctg tcctgcagag cctcccagtc cgtgacctcc 1500 tcctacctcg cctggtatca gcagaagccc ggccaggccc ctcggctgct gatcaacgtg 1560 ggcagtcgga gagccaccgg catccctgac cggttctccg gctctggctc cggcaccgac 1620 ttcaccctga ccatctcccg gctggaaccc gaggacttcg ccgtgtacta ctgccagcag 1680 ggcatcatgc tgccccccac ctttggccag ggcaccaagg tggaaatcaa g 1731 <![CDATA[<210> 330]]> <![CDATA[<211> 586]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (9B11) VHCH1 Fc杵VH (4B9)]]> <![CDATA[<400> 330]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu 465 470 475 480 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 485 490 495 Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys 500 505 510 Gly Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr 515 520 525 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser 530 535 540 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 545 550 555 560 Ala Val Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp 565 570 575 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 580 585 <![CDATA[<210> 331]]> <![CDATA[<211> 577]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (9B11) VHCH1 Fc臼VL (4B9)]]> <![CDATA[<400> 331]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu 465 470 475 480 Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln 485 490 495 Ser Val Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 500 505 510 Ala Pro Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile 515 520 525 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 530 535 540 Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln 545 550 555 560 Gly Ile Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 565 570 575 Lys <![CDATA[<210> 332]]> <![CDATA[<211> 1731]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (12B3) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 332]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctggagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcgagt 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccctg cagagatgag 1080 ctgaccaaga accaggtgtc cctgtggtgt ctggtcaagg gcttctaccc cagcgatatc 1140 gccgtggagt gggagagcaa cggccagcct gagaacaact acaagaccac cccccctgtg 1200 ctggacagcg acggcagctt cttcctgtac tccaaactga ccgtggacaa gagccggtgg 1260 cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagtccc tgagcctgag ccccggcgga ggcggcggaa gcggaggagg aggatctggg 1380 ggcggaggtt ccggaggcgg tggatctgag atcgtgctga cccagtctcc cggcaccctg 1440 tctctgagcc ctggcgagag agccaccctg tcctgcagag cctcccagtc cgtgacctcc 1500 tcctacctcg cctggtatca gcagaagccc ggccaggccc ctcggctgct gatcaacgtg 1560 ggcagtcgga gagccaccgg catccctgac cggttctccg gctctggctc cggcaccgac 1620 ttcaccctga ccatctcccg gctggaaccc gaggacttcg ccgtgtacta ctgccagcag 1680 ggcatcatgc tgccccccac ctttggccag ggcaccaagg tggaaatcaa g 1731 <![CDATA[<210> 333]]> <![CDATA[<211> 1758]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (12B3) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 333]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctggagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctgaa 300 ttccgtttct acgctgactt cgactactgg ggccaaggga ccaccgtgac cgtctcgagt 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtgcaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctctcgtgc gcagtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctcgtg agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtgga ggcggcggaa gcggaggagg aggatccggc 1380 ggcggaggtt ccggaggcgg aggatccgag gtgcagctgc tcgaaagcgg cggaggactg 1440 gtgcagcctg gcggcagcct gagactgtct tgcgccgcca gcggcttcac cttcagcagc 1500 tacgccatga gctgggtccg ccaggcccct ggcaagggac tggaatgggt gtccgccatc 1560 atcggctctg gcgccagcac ctactacgcc gacagcgtga agggccggtt caccatcagc 1620 cgggacaaca gcaagaacac cctgtacctg cagatgaaca gcctgcgggc cgaggacacc 1680 gccgtgtact actgcgccaa gggatggttc ggcggcttca actactgggg acagggcacc 1740 ctggtcaccg tgtccagc 1758 <![CDATA[<210> 334]]> <![CDATA[<211> 577]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (12B3) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 334]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu 465 470 475 480 Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln 485 490 495 Ser Val Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 500 505 510 Ala Pro Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile 515 520 525 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 530 535 540 Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln 545 550 555 560 Gly Ile Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 565 570 575 Lys <![CDATA[<210> 335]]> <![CDATA[<211> 586]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (12B3) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 335]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Glu Phe Arg Phe Tyr Ala Asp Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu 465 470 475 480 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 485 490 495 Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys 500 505 510 Gly Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr 515 520 525 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser 530 535 540 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 545 550 555 560 Ala Val Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp 565 570 575 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 580 585 <![CDATA[<210> 336]]> <![CDATA[<211> 1725]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (25G7) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 336]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgacgac 300 ccgtggccgc cgttcgacta ctggggccaa ggaaccctgg tcaccgtctc gagtgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtac accctgcccc cctgcagaga tgagctgacc 1080 aagaaccagg tgtccctgtg gtgtctggtc aagggcttct accccagcga tatcgccgtg 1140 gagtgggaga gcaacggcca gcctgagaac aactacaaga ccaccccccc tgtgctggac 1200 agcgacggca gcttcttcct gtactccaaa ctgaccgtgg acaagagccg gtggcagcag 1260 ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta cacccagaag 1320 tccctgagcc tgagccccgg cggaggcggc ggaagcggag gaggaggatc tgggggcgga 1380 ggttccggag gcggtggatc tgagatcgtg ctgacccagt ctcccggcac cctgtctctg 1440 agccctggcg agagagccac cctgtcctgc agagcctccc agtccgtgac ctcctcctac 1500 ctcgcctggt atcagcagaa gcccggccag gcccctcggc tgctgatcaa cgtgggcagt 1560 cggagagcca ccggcatccc tgaccggttc tccggctctg gctccggcac cgacttcacc 1620 ctgaccatct cccggctgga acccgaggac ttcgccgtgt actactgcca gcagggcatc 1680 atgctgcccc ccacctttgg ccagggcacc aaggtggaaa tcaag 1725 <![CDATA[<210> 337]]> <![CDATA[<211> 1752]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (25G7) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 337]]> gaggtgcaat tgttggagtc tgggggaggc ttggtacagc ctggggggtc cctgagactc 60 tcctgtgcag cctccggatt cacctttagc agttatgcca tgagctgggt ccgccaggct 120 ccagggaagg ggctggagtg ggtctcagct attagtggta gtggtggtag cacatactac 180 gcagactccg tgaagggccg gttcaccatc tccagagaca attccaagaa cacgctgtat 240 ctgcagatga acagcctgag agccgaggac acggccgtat attactgtgc gcgtgacgac 300 ccgtggccgc cgttcgacta ctggggccaa ggaaccctgg tcaccgtctc gagtgctagc 360 accaagggcc catcggtctt ccccctggca ccctcctcca agagcacctc tgggggcaca 420 gcggccctgg gctgcctggt caaggactac ttccccgaac cggtgacggt gtcgtggaac 480 tcaggcgccc tgaccagcgg cgtgcacacc ttcccggctg tcctacagtc ctcaggactc 540 tactccctca gcagcgtggt gaccgtgccc tccagcagct tgggcaccca gacctacatc 600 tgcaacgtga atcacaagcc cagcaacacc aaggtggaca agaaagttga gcccaaatct 660 tgtgacaaaa ctcacacatg cccaccgtgc ccagcacctg aagctgcagg gggaccgtca 720 gtcttcctct tccccccaaa acccaaggac accctcatga tctcccggac ccctgaggtc 780 acatgcgtgg tggtggacgt gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg 840 gacggcgtgg aggtgcataa tgccaagaca aagccgcggg aggagcagta caacagcacg 900 taccgtgtgg tcagcgtcct caccgtcctg caccaggact ggctgaatgg caaggagtac 960 aagtgcaagg tctccaacaa agccctcggc gcccccatcg agaaaaccat ctccaaagcc 1020 aaagggcagc cccgagaacc acaggtgtgc accctgcccc catcccggga tgagctgacc 1080 aagaaccagg tcagcctctc gtgcgcagtc aaaggcttct atcccagcga catcgccgtg 1140 gagtgggaga gcaatgggca gccggagaac aactacaaga ccacgcctcc cgtgctggac 1200 tccgacggct ccttcttcct cgtgagcaag ctcaccgtgg acaagagcag gtggcagcag 1260 gggaacgtct tctcatgctc cgtgatgcat gaggctctgc acaaccacta cacgcagaag 1320 agcctctccc tgtctccggg tggaggcggc ggaagcggag gaggaggatc cggcggcgga 1380 ggttccggag gcggaggatc cgaggtgcag ctgctcgaaa gcggcggagg actggtgcag 1440 cctggcggca gcctgagact gtcttgcgcc gccagcggct tcaccttcag cagctacgcc 1500 atgagctggg tccgccaggc ccctggcaag ggactggaat gggtgtccgc catcatcggc 1560 tctggcgcca gcacctacta cgccgacagc gtgaagggcc ggttcaccat cagccgggac 1620 aacagcaaga acaccctgta cctgcagatg aacagcctgc gggccgagga caccgccgtg 1680 tactactgcg ccaagggatg gttcggcggc ttcaactact ggggacaggg caccctggtc 1740 accgtgtcca gc 1752 <![CDATA[<210> 338]]> <![CDATA[<211> 575]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (25G7) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 338]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu 340 345 350 Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp Cys 355 360 365 Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu 465 470 475 480 Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val 485 490 495 Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro 500 505 510 Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile Pro Asp 515 520 525 Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 530 535 540 Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly Ile 545 550 555 560 Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 339]]> <![CDATA[<211> 584]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (25G7) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 339]]> Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 1 5 10 15 Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 20 25 30 Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45 Ser Ala Ile Ser Gly Ser Gly Gly Ser Thr Tyr Tyr Ala Asp Ser Val 50 55 60 Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 65 70 75 80 Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Asp Asp Pro Trp Pro Pro Phe Asp Tyr Trp Gly Gln Gly Thr 100 105 110 Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro 115 120 125 Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly 130 135 140 Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn 145 150 155 160 Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln 165 170 175 Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser 180 185 190 Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser 195 200 205 Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr 210 215 220 His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro Ser 225 230 235 240 Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg 245 250 255 Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro 260 265 270 Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala 275 280 285 Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val 290 295 300 Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr 305 310 315 320 Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys Thr 325 330 335 Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr Leu 340 345 350 Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser Cys 355 360 365 Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser 370 375 380 Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp 385 390 395 400 Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys Ser 405 410 415 Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala 420 425 430 Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Gly 435 440 445 Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly 450 455 460 Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln 465 470 475 480 Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe 485 490 495 Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu 500 505 510 Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr Ala 515 520 525 Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn 530 535 540 Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val 545 550 555 560 Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly Gln 565 570 575 Gly Thr Leu Val Thr Val Ser Ser 580 <![CDATA[<210> 340]]> <![CDATA[<211> 1728]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (11D5) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 340]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctact 300 ctgatctacg gttacttcga ctactggggc caagggacca ccgtgaccgt ctcctcagct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaagctgc agggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ggcgccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tacaccctgc ccccctgcag agatgagctg 1080 accaagaacc aggtgtccct gtggtgtctg gtcaagggct tctaccccag cgatatcgcc 1140 gtggagtggg agagcaacgg ccagcctgag aacaactaca agaccacccc ccctgtgctg 1200 gacagcgacg gcagcttctt cctgtactcc aaactgaccg tggacaagag ccggtggcag 1260 cagggcaacg tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag 1320 aagtccctga gcctgagccc cggcggaggc ggcggaagcg gaggaggagg atctgggggc 1380 ggaggttccg gaggcggtgg atctgagatc gtgctgaccc agtctcccgg caccctgtct 1440 ctgagccctg gcgagagagc caccctgtcc tgcagagcct cccagtccgt gacctcctcc 1500 tacctcgcct ggtatcagca gaagcccggc caggcccctc ggctgctgat caacgtgggc 1560 agtcggagag ccaccggcat ccctgaccgg ttctccggct ctggctccgg caccgacttc 1620 accctgacca tctcccggct ggaacccgag gacttcgccg tgtactactg ccagcagggc 1680 atcatgctgc cccccacctt tggccagggc accaaggtgg aaatcaag 1728 <![CDATA[<210> 341]]> <![CDATA[<211> 1755]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (11D5) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 341]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatctact 300 ctgatctacg gttacttcga ctactggggc caagggacca ccgtgaccgt ctcctcagct 360 agcaccaagg gcccatcggt cttccccctg gcaccctcct ccaagagcac ctctgggggc 420 acagcggccc tgggctgcct ggtcaaggac tacttccccg aaccggtgac ggtgtcgtgg 480 aactcaggcg ccctgaccag cggcgtgcac accttcccgg ctgtcctaca gtcctcagga 540 ctctactccc tcagcagcgt ggtgaccgtg ccctccagca gcttgggcac ccagacctac 600 atctgcaacg tgaatcacaa gcccagcaac accaaggtgg acaagaaagt tgagcccaaa 660 tcttgtgaca aaactcacac atgcccaccg tgcccagcac ctgaagctgc agggggaccg 720 tcagtcttcc tcttcccccc aaaacccaag gacaccctca tgatctcccg gacccctgag 780 gtcacatgcg tggtggtgga cgtgagccac gaagaccctg aggtcaagtt caactggtac 840 gtggacggcg tggaggtgca taatgccaag acaaagccgc gggaggagca gtacaacagc 900 acgtaccgtg tggtcagcgt cctcaccgtc ctgcaccagg actggctgaa tggcaaggag 960 tacaagtgca aggtctccaa caaagccctc ggcgccccca tcgagaaaac catctccaaa 1020 gccaaagggc agccccgaga accacaggtg tgcaccctgc ccccatcccg ggatgagctg 1080 accaagaacc aggtcagcct ctcgtgcgca gtcaaaggct tctatcccag cgacatcgcc 1140 gtggagtggg agagcaatgg gcagccggag aacaactaca agaccacgcc tcccgtgctg 1200 gactccgacg gctccttctt cctcgtgagc aagctcaccg tggacaagag caggtggcag 1260 caggggaacg tcttctcatg ctccgtgatg catgaggctc tgcacaacca ctacacgcag 1320 aagagcctct ccctgtctcc gggtggaggc ggcggaagcg gaggaggagg atccggcggc 1380 ggaggttccg gaggcggagg atccgaggtg cagctgctcg aaagcggcgg aggactggtg 1440 cagcctggcg gcagcctgag actgtcttgc gccgccagcg gcttcacctt cagcagctac 1500 gccatgagct gggtccgcca ggcccctggc aagggactgg aatgggtgtc cgccatcatc 1560 ggctctggcg ccagcaccta ctacgccgac agcgtgaagg gccggttcac catcagccgg 1620 gacaacagca agaacaccct gtacctgcag atgaacagcc tgcgggccga ggacaccgcc 1680 gtgtactact gcgccaaggg atggttcggc ggcttcaact actggggaca gggcaccctg 1740 gtcaccgtgt ccagc 1755 <![CDATA[<210> 342]]> <![CDATA[<211> 576]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (11D5) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 342]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr 340 345 350 Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Trp 355 360 365 Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 450 455 460 Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser 465 470 475 480 Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser 485 490 495 Val Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala 500 505 510 Pro Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile Pro 515 520 525 Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 530 535 540 Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Gly 545 550 555 560 Ile Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 565 570 575 <![CDATA[<210> 343]]> <![CDATA[<211> 585]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (11D5) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 343]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Thr Leu Ile Tyr Gly Tyr Phe Asp Tyr Trp Gly Gln Gly 100 105 110 Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe 115 120 125 Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu 130 135 140 Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp 145 150 155 160 Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu 165 170 175 Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser 180 185 190 Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro 195 200 205 Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys 210 215 220 Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly Pro 225 230 235 240 Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser 245 250 255 Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu Asp 260 265 270 Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn 275 280 285 Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val 290 295 300 Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu 305 310 315 320 Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu Lys 325 330 335 Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys Thr 340 345 350 Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu Ser 355 360 365 Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu 370 375 380 Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu 385 390 395 400 Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp Lys 405 410 415 Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu 420 425 430 Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly 435 440 445 Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly 450 455 460 Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val 465 470 475 480 Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr 485 490 495 Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly 500 505 510 Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr Tyr 515 520 525 Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys 530 535 540 Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala 545 550 555 560 Val Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp Gly 565 570 575 Gln Gly Thr Leu Val Thr Val Ser Ser 580 585 <![CDATA[<210> 344]]> <![CDATA[<211> 1731]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (9B11) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 344]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtacaccc tgcccccctg cagagatgag 1080 ctgaccaaga accaggtgtc cctgtggtgt ctggtcaagg gcttctaccc cagcgatatc 1140 gccgtggagt gggagagcaa cggccagcct gagaacaact acaagaccac cccccctgtg 1200 ctggacagcg acggcagctt cttcctgtac tccaaactga ccgtggacaa gagccggtgg 1260 cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1320 cagaagtccc tgagcctgag ccccggcgga ggcggcggaa gcggaggagg aggatctggg 1380 ggcggaggtt ccggaggcgg tggatctgag atcgtgctga cccagtctcc cggcaccctg 1440 tctctgagcc ctggcgagag agccaccctg tcctgcagag cctcccagtc cgtgacctcc 1500 tcctacctcg cctggtatca gcagaagccc ggccaggccc ctcggctgct gatcaacgtg 1560 ggcagtcgga gagccaccgg catccctgac cggttctccg gctctggctc cggcaccgac 1620 ttcaccctga ccatctcccg gctggaaccc gaggacttcg ccgtgtacta ctgccagcag 1680 ggcatcatgc tgccccccac ctttggccag ggcaccaagg tggaaatcaa g 1731 <![CDATA[<210> 345]]> <![CDATA[<211> 1758]]> <![CDATA[<212> DNA]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> DNA (9B11) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 345]]> caggtgcaat tggtgcagtc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc 60 tcctgcaagg cctccggagg cacattcagc agctacgcta taagctgggt gcgacaggcc 120 cctggacaag ggctcgagtg gatgggaggg atcatcccta tctttggtac agcaaactac 180 gcacagaagt tccagggcag ggtcaccatt actgcagaca aatccacgag cacagcctac 240 atggagctga gcagcctgag atctgaggac accgccgtgt attactgtgc gagatcttct 300 ggtgcttacc cgggttactt cgactactgg ggccaaggga ccaccgtgac cgtctcctca 360 gctagcacca agggcccatc ggtcttcccc ctggcaccct cctccaagag cacctctggg 420 ggcacagcgg ccctgggctg cctggtcaag gactacttcc ccgaaccggt gacggtgtcg 480 tggaactcag gcgccctgac cagcggcgtg cacaccttcc cggctgtcct acagtcctca 540 ggactctact ccctcagcag cgtggtgacc gtgccctcca gcagcttggg cacccagacc 600 tacatctgca acgtgaatca caagcccagc aacaccaagg tggacaagaa agttgagccc 660 aaatcttgtg acaaaactca cacatgccca ccgtgcccag cacctgaagc tgcaggggga 720 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct 780 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg 840 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac 900 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag 960 gagtacaagt gcaaggtctc caacaaagcc ctcggcgccc ccatcgagaa aaccatctcc 1020 aaagccaaag ggcagccccg agaaccacag gtgtgcaccc tgcccccatc ccgggatgag 1080 ctgaccaaga accaggtcag cctctcgtgc gcagtcaaag gcttctatcc cagcgacatc 1140 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg 1200 ctggactccg acggctcctt cttcctcgtg agcaagctca ccgtggacaa gagcaggtgg 1260 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg 1320 cagaagagcc tctccctgtc tccgggtgga ggcggcggaa gcggaggagg aggatccggc 1380 ggcggaggtt ccggaggcgg aggatccgag gtgcagctgc tcgaaagcgg cggaggactg 1440 gtgcagcctg gcggcagcct gagactgtct tgcgccgcca gcggcttcac cttcagcagc 1500 tacgccatga gctgggtccg ccaggcccct ggcaagggac tggaatgggt gtccgccatc 1560 atcggctctg gcgccagcac ctactacgcc gacagcgtga agggccggtt caccatcagc 1620 cgggacaaca gcaagaacac cctgtacctg cagatgaaca gcctgcgggc cgaggacacc 1680 gccgtgtact actgcgccaa gggatggttc ggcggcttca actactgggg acagggcacc 1740 ctggtcaccg tgtccagc 1758 <![CDATA[<210> 346]]> <![CDATA[<211> 577]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (9B11) VHCH1 Fc杵VL (4B9)]]> <![CDATA[<400> 346]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 340 345 350 Thr Leu Pro Pro Cys Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Trp Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu 465 470 475 480 Ser Leu Ser Pro Gly Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln 485 490 495 Ser Val Thr Ser Ser Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln 500 505 510 Ala Pro Arg Leu Leu Ile Asn Val Gly Ser Arg Arg Ala Thr Gly Ile 515 520 525 Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 530 535 540 Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln 545 550 555 560 Gly Ile Met Leu Pro Pro Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 565 570 575 Lys <![CDATA[<210> 347]]> <![CDATA[<211> 586]]> <![CDATA[<212> PRT]]> <![CDATA[<213> 人工序列]]> <![CDATA[<220>]]> <![CDATA[<223> (9B11) VHCH1 Fc臼VH (4B9)]]> <![CDATA[<400> 347]]> Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 1 5 10 15 Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 20 25 30 Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 35 40 45 Gly Gly Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 50 55 60 Gln Gly Arg Val Thr Ile Thr Ala Asp Lys Ser Thr Ser Thr Ala Tyr 65 70 75 80 Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Ser Ser Gly Ala Tyr Pro Gly Tyr Phe Asp Tyr Trp Gly Gln 100 105 110 Gly Thr Thr Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 115 120 125 Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 130 135 140 Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 145 150 155 160 Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 165 170 175 Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 180 185 190 Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 195 200 205 Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 210 215 220 Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Ala Ala Gly Gly 225 230 235 240 Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 245 250 255 Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 260 265 270 Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 275 280 285 Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 290 295 300 Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 305 310 315 320 Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Gly Ala Pro Ile Glu 325 330 335 Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Cys 340 345 350 Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys Asn Gln Val Ser Leu 355 360 365 Ser Cys Ala Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 370 375 380 Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 385 390 395 400 Leu Asp Ser Asp Gly Ser Phe Phe Leu Val Ser Lys Leu Thr Val Asp 405 410 415 Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 420 425 430 Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 435 440 445 Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser 450 455 460 Gly Gly Gly Gly Ser Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu 465 470 475 480 Val Gln Pro Gly Gly Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe 485 490 495 Thr Phe Ser Ser Tyr Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys 500 505 510 Gly Leu Glu Trp Val Ser Ala Ile Ile Gly Ser Gly Ala Ser Thr Tyr 515 520 525 Tyr Ala Asp Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser 530 535 540 Lys Asn Thr Leu Tyr Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr 545 550 555 560 Ala Val Tyr Tyr Cys Ala Lys Gly Trp Phe Gly Gly Phe Asn Tyr Trp 565 570 575 Gly Gln Gly Thr Leu Val Thr Val Ser Ser 580 585
Claims (5)
- 一種特異性結合於OX40之抗體,其中該抗體包含 (i) 包含胺基酸序列SEQ ID NO:25之重鏈可變區VH及包含胺基酸序列SEQ ID NO:26之輕鏈可變區VL, (ii) 包含胺基酸序列SEQ ID NO:27之重鏈可變區VH及包含胺基酸序列SEQ ID NO:28之輕鏈可變區VL, (iii) 包含胺基酸序列SEQ ID NO:29之重鏈可變區VH及包含胺基酸序列SEQ ID NO:30之輕鏈可變區VL, (iv) 包含胺基酸序列SEQ ID NO:31之重鏈可變區VH及包含胺基酸序列SEQ ID NO:32之輕鏈可變區VL, (v) 包含胺基酸序列SEQ ID NO:33之重鏈可變區VH及包含胺基酸序列SEQ ID NO:34之輕鏈可變區VL, (vi) 包含胺基酸序列SEQ ID NO:35之重鏈可變區VH及包含胺基酸序列SEQ ID NO:36之輕鏈可變區VL,或 (vii) 包含胺基酸序列SEQ ID NO:37之重鏈可變區VH及包含胺基酸序列SEQ ID NO:38之輕鏈可變區VL。
- 一種聚核苷酸,其編碼如請求項1之抗體。
- 一種醫藥組合物,其包含如請求項1之抗體及至少一種醫藥學上可接受之賦形劑。
- 如請求項3之醫藥組合物,其用作醫藥品。
- 一種如請求項1之抗體用於製備醫藥品之用途,其中該醫藥品用於: (i) 刺激T細胞反應, (ii) 支持活化T細胞之存活, (iii) 治療感染, (iv) 治療癌症, (v) 延遲癌症進展,或 (vi) 延長罹患癌症之患者的存活期。
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US20090162359A1 (en) | 2007-12-21 | 2009-06-25 | Christian Klein | Bivalent, bispecific antibodies |
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EP2681239B8 (en) | 2011-02-28 | 2015-09-09 | F. Hoffmann-La Roche AG | Antigen binding proteins |
MX342034B (es) | 2011-02-28 | 2016-09-12 | Hoffmann La Roche | Proteinas monovalentes que se unen a antigenos. |
JP6422956B2 (ja) * | 2013-10-11 | 2018-11-14 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | 多重特異性ドメイン交換共通可変軽鎖抗体 |
PL3224275T3 (pl) | 2014-11-14 | 2020-09-07 | F.Hoffmann-La Roche Ag | Cząsteczki wiążące antygen zawierające trimer liganda z rodziny TNF |
US11566082B2 (en) | 2014-11-17 | 2023-01-31 | Cytiva Bioprocess R&D Ab | Mutated immunoglobulin-binding polypeptides |
WO2016156291A1 (en) | 2015-03-31 | 2016-10-06 | F. Hoffmann-La Roche Ag | Antigen binding molecules comprising a trimeric tnf family ligand |
AR106188A1 (es) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | Anticuerpos anti-cd19 humano humanizados y métodos de utilización |
MA43017A (fr) | 2015-10-02 | 2018-08-08 | Hoffmann La Roche | Anticorps bispécifiques spécifiques d'un récepteur de co-stimulation du tnf |
US10730908B2 (en) | 2016-05-11 | 2020-08-04 | Ge Healthcare Bioprocess R&D Ab | Separation method |
US10889615B2 (en) | 2016-05-11 | 2021-01-12 | Cytiva Bioprocess R&D Ab | Mutated immunoglobulin-binding polypeptides |
US10703774B2 (en) | 2016-09-30 | 2020-07-07 | Ge Healthcare Bioprocess R&D Ab | Separation method |
US10654887B2 (en) | 2016-05-11 | 2020-05-19 | Ge Healthcare Bio-Process R&D Ab | Separation matrix |
ES2909833T3 (es) | 2016-05-11 | 2022-05-10 | Cytiva Bioprocess R & D Ab | Método de limpieza y/o desinfección de una matriz de separación |
CN109311949B (zh) | 2016-05-11 | 2022-09-16 | 思拓凡生物工艺研发有限公司 | 储存分离基质的方法 |
ES2874974T3 (es) | 2016-05-11 | 2021-11-05 | Cytiva Bioprocess R & D Ab | Matriz de separación |
KR102632202B1 (ko) | 2016-07-14 | 2024-02-02 | 젠맵 에이/에스 | Cd40 및 cd137에 대한 다중특이적 항체 |
WO2018027204A1 (en) | 2016-08-05 | 2018-02-08 | Genentech, Inc. | Multivalent and multiepitopic anitibodies having agonistic activity and methods of use |
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