TW202003037A - 引導及導航控制蛋白及其製備和使用方法 - Google Patents
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Abstract
本申請案提供引導及導航控制(GNC)蛋白質。在一個實施例中,該引導及導航控制(GNC)蛋白質包含用於T細胞活化受體之結合結構域、用於腫瘤相關聯抗原之結合結構域、用於免疫查核點受體之結合結構域、及用於T細胞共刺激受體之結合結構域。該用於該腫瘤相關聯抗原之結合結構域不相鄰於該用於該T細胞共刺激受體之結合結構域。在一個實施例中,該用於該T細胞活化受體之結合結構域相鄰於該用於該腫瘤相關聯抗原(TAA)之結合結構域。
Description
本申請案主張2018年3月27日申請的美國臨時專利申請案第62/648880號及2018年3月27日申請的美國臨時專利申請案第62/648888號之權益,其全部揭示內容係以引用方式明確併入本文。
本申請案大體上係關於引導及導航控制(Guidance and Navigation Control;GNC)蛋白質之技術領域,該等蛋白質具有針對免疫細胞及腫瘤細胞上的表面分子之多特異性結合活性,且更特定而言本申請案係關於製造及使用GNC蛋白質。
癌細胞發展各種策略來逃避免疫系統。用於免疫逃脫的隱伏機制之一係減少癌細胞藉由免疫系統之識別。癌症特異性抗原之缺陷性呈現或其缺乏造成免疫耐受性及癌症進程。在存在有效免疫識別的情況下,腫瘤使用其他機制來避免藉由免疫系統消除。免疫活性腫瘤產生抑制性微環境來下調免疫反應。在成形抑制性腫瘤微環境方面涉及多個參與者,包括腫瘤細胞、調節T細胞、骨髓源性抑制性細胞、基質細胞、及其他細胞類型。免疫反應之抑制可以細胞接觸獨立性方式,經由免疫抑制細胞介素之分泌或消除來自局部環境的必要存活因子來執行。細胞接觸依賴性抑制依賴於細胞表面上表現的分子,例如程序性死亡配位體1 (Programmed Death Ligand 1;PD-L1)、T-淋巴細胞相關聯蛋白質4 (CTLA-4)及其他[Dunn等人, 2004, Immunity, 21(2): 137-48; Adachi & Tamada, 2015, Cancer Sci., 106(8): 945-50]。
由於腫瘤逃避藉由免疫系統識別的機制持續得以更好地理解,最近已出現靶向該等機制之新治療模態。在2011年3月25日,美國食品藥物管理局(Food and Drug Administration;FDA)批准依匹單抗注射(Yervoy, Bristol-Myers Myers Squibb)用於治療不可切除的或轉移性黑素瘤。Yervoy結合至活化T細胞上表現的細胞毒性T-淋巴細胞相關聯蛋白質4 (CTLA-4)且阻斷CTLA-4與抗原呈遞細胞上之CD80/86之相互作用,進而阻斷負或抑制信號透過CTLA-4遞送至T細胞中,引起抗原特異性T細胞之重新活化,從而在許多患者中導致腫瘤之根除。幾年後,在2014年,FDA批准的Keytruda (派姆單抗,Merck)及Opdivo (納武單抗,Bristol-Myers Squibb)用於治療晚期黑素瘤。該等單株抗體結合至在活化及/或耗盡的T細胞上表現的PD-1且阻斷PD-1與腫瘤上表現的PD-L1之相互作用,進而消除抑制信號透過PD-1進入T細胞中,引起抗原特異性T細胞之重新活化,又在許多患者中導致腫瘤之根除。自那時起,已執行另外的臨床試驗來在晚期黑素瘤之治療中比較單一單株抗體Yervoy與單株抗體Yervoy及Opdivo之組合,從而證實在利用抗體之組合治療的患者中在總體存活及無進展存活方面的改良。(Hodi等人, 2016, Lancet Oncol. 17(11):1558-1568,Hellman等人, 2018, Cancer Cell 33(5): 853-861)。然而,由於許多臨床試驗已證實利用對一或多個免疫查核點分子為特異性的單株抗體治療癌症患者之巨大益處,資料已顯露僅具有高突變負荷之彼等患者才展示臨床反應,該高突變負荷產生藉由抗原特異性T細胞識別的新型T細胞表位(Snyder等人, 2014, NEJM 371:2189-2199)。具有低腫瘤突變負荷之彼等患者幾乎不展示目標臨床反應(Snyder等人, 2014, NEJM 371:2189-2199,Hellman等人, 2018, Cancer Cell 33(5): 853-861)。
近年來,其他團體已開發替代方法,該方法不需要藉由抗原呈現細胞向活化T細胞的新生表位呈現之存在。一個實例係雙特異性抗體之開發,其中抗體的對例如CD19之腫瘤相關聯抗原為特異性的結合結構域係連接至對T細胞上之CD3為特異性的抗體結合結構域,因而產生雙特異性T細胞銜接子(engager)或BiTe分子。在2014年,FDA批准稱為博納吐單抗之雙特異性抗體用於治療前驅B細胞急性淋巴母細胞性白血病。博納吐單抗將對白血病細胞上表現的CD19為特異性的scFv與對T細胞上表現的CD3為特異性的scFv連接(Bejnjamin及Stein 2016, Ther Adv Hematol 7 (3): 142-146)。然而,儘管在患有復發或難治性ALL的患者中有>50%之初始反應速率,但許多患者在利用博納吐單抗成功治療之後對博納吐單抗療法有抵抗力或復發。正在出現的證據是,對博納吐單抗有抵抗力或在博納吐單抗治療之後復發的患者可歸因於在腫瘤細胞上表現的免疫查核點抑制分子之表現,該等免疫查核點抑制分子諸如PD-L1,其驅動抑制信號透過活化T細胞上表現的PD-1 (Feucht等人, 2016, Oncotarget 7(47):76902-76919)。在對利用博納吐單抗之療法有抵抗力的患者之病例研究中,執行第二輪博納吐單抗療法但添加單株抗體派姆單抗(Keytruda, Merck),其特異地結合至PD-1且阻斷表現T細胞之PD-1與腫瘤細胞表現之PD-L1之相互作用,在此一位患者中引起顯著的反應及在骨髓中腫瘤細胞自45%至小於5%之減少(Feucht等人, 2016, Oncotarget 7(47):76902-76919)。該些結果證實,將雙特異性BiTe分子與一或多個單株抗體組合可相較於任一藥劑單獨而言顯著地增加臨床活性。儘管存在有前景的結果,但進行組合療法之成本必定是高的,此歸因於在招募的代表性群體中的多次臨床試驗及困難度。
利用嵌合抗原受體T細胞(chimeric antigen receptor T cell;CAR-T)之授受性細胞療法係用於治療癌症的另一有前景免疫療法。CAR-T療法之臨床成功已揭露患有CD19陽性、治療難治性B細胞惡性腫瘤之患者的耐久完全緩解及延長的存活(Gill & June. 2015. Immunol Rev, 263: 68-89)。然而,與個別化及遺傳修飾CAR-T免疫療法之製造相關聯的成本及複雜性已將其產生及使用限制於用於治療相對小數量的患者之專業中心。亦稱為細胞介素風暴的細胞介素釋放症候群(cytokine release syndrome;CRS)係在工程化CAR-T細胞之輸注之後的最顯著不利效應(Bonifant等人, 201, Mol Ther Oncolytics. 3: 16011)。在許多情況下,CRS之發病及嚴重程度似乎是專門的個人事件。緩和CRS之當前選擇主要地集中在快速反應及管理護理,因為在T細胞輸注之前控制CRS之選擇是有限的。
儘管現已建立對CD19陽性B細胞惡性腫瘤為特異性的CAR-T療法之功效,但CAR-T療法針對實體腫瘤之功效迄今為止尚未得到明確證明。當前,許多臨床試驗正在進行中以探究用於CAR-T療法之各種實體腫瘤相關聯抗原(tumor-associated antigen;TAA)。T細胞低效地轉運至腫瘤中、免疫抑制腫瘤微環境、次最佳抗原識別特異性、及缺乏對治療有關不利事件之控制當前係視為實體腫瘤CAR-T療法中之主要障礙(Li等人, 2018, J Hematol Oncol. 11(1):22-40)。管控治療效應以及CAR-T細胞輸注之前及之後的任何不利效應之選擇是有限的。
本申請案提供對T細胞及腫瘤細胞之表面分子具有多特異性抗原結合活性的引導及導航控制(guidance and navigation control;GNC)蛋白質。在一個實施例中,引導及導航控制(GNC)蛋白質包含用於T細胞活化受體之結合結構域、用於腫瘤相關聯抗原之結合結構域、用於免疫查核點受體之結合結構域、及用於T細胞共刺激受體之結合結構域。
在一個實施例中,用於腫瘤相關聯抗原之結合結構域不相鄰於用於T細胞共刺激受體之結合結構域。在一個實施例中,用於T細胞活化受體之結合結構域相鄰於用於腫瘤相關聯抗原(tumor associated antigen;TAA)之結合結構域。T細胞活化受體可包括而不限於CD3。T細胞共刺激受體可包括而不限於4-1BB、CD28、OX40、GITR、CD40L、ICOS、Light、CD27、CD30、或其組合。免疫查核點受體可包括而不限於PD-L1、PD-1、TIGIT、TIM-3、LAG-3、CTLA4、BTLA、VISTA、PDL2、CD160、LOX-1、siglec-15、CD47、或其組合。
腫瘤相關聯抗原(tumor associated antigen;TAA)可包括而不限於ROR1、CD19、EGFRVIII、BCMA、CD20、CD33、CD123、CD22、CD30、CEA、HER2、EGFR、LMP1、LMP2A、間皮素、PSMA、EpCAM、磷脂醯肌醇蛋白聚糖-3、gpA33、GD2、TROP2、或其組合。在一個實施例中,腫瘤相關聯抗原可為ROR1。在一個實施例中,腫瘤相關聯抗原可為CD19。在一個實施例中,腫瘤相關聯抗原可為EGFRVIII。
在一個實施例中,腫瘤相關聯抗原可為以下各項之上的受體:肺癌細胞、肝癌細胞、乳癌細胞、結腸直腸癌細胞、肛門癌細胞、胰腺癌細胞、膽囊癌細胞、膽管癌細胞、頭頸癌細胞、鼻咽癌細胞、皮膚癌細胞、黑素瘤細胞、卵巢癌細胞、前列腺癌細胞、尿道癌細胞、肺癌細胞、非小肺細胞癌細胞、小細胞肺癌細胞、腦腫瘤細胞、神經膠質瘤細胞、神經胚細胞瘤細胞、食道癌細胞、胃癌細胞、肝癌細胞、腎癌細胞、膀胱癌細胞、子宮頸癌細胞、子宮內膜癌細胞、甲狀腺癌細胞、眼癌細胞、肉瘤細胞、骨癌細胞、白血病細胞、骨髓瘤細胞、淋巴瘤細胞、或其組合。在一個實施例中,腫瘤相關聯抗原可為B細胞上之受體。
在一個實施例中,引導及導航控制(guidance and navigation control;GNC)蛋白質可為抗體或抗體單體或其片段。在一個實施例中,GNC蛋白質可為三特異性抗體。在一個實施例中,GNC蛋白質可為四特異性抗體。在一個實施例中,GNC蛋白質包括Fc結構域或其片段。可使用來自抗體之任何Fc結構域。示例性Fc結構域可包括來自IgG、IgA、IgD、IgM、IgE、或其片段或組合的Fc結構域。Fc結構域可為天然的或工程化的。在一個實施例中,Fc結構域可含有抗原結合位點。
在一個實施例中,引導及導航控制(GNC)蛋白質為抗體。在一個實施例中,腫瘤相關聯抗原包含ROR1、CD19、或EGRFVIII。在一個實施例中,T細胞活化受體包含CD3且用於CD3之結合結構域可經由連接子連接至用於腫瘤相關聯抗原(tumor associated;TAA)的結合結構域以形成CD3-TAA對。在一個實施例中,IgG Fc結構域可中介CD3-TAA對及用於免疫查核點受體之結合結構域。在一個實施例中,免疫查核點受體可為PD-L1。
在一個實施例中,連接子可為共價鍵。在一個實施例中,連接子可為肽連接子。在一個實施例中,肽連接子具有不超過100個胺基酸之長度。在一個實施例中,肽連接子具有不超過2、5、10、20、30、40、50、60、70、80、90或100個胺基酸之長度。在一個實施例中,肽連接子具有不超過10個胺基酸之長度。在一個實施例中,肽連接子具有約2個胺基酸至約10個胺基酸之長度。在一個實施例中,肽連接子包括2、5、或10個胺基酸。
在一個實施例中,引導及導航控制(GNC)蛋白質具有N末端及C末端,其自N末端至C末端串聯地包含用於CD3之結合結構域、用於EGFRVIII之結合結構域、IgG Fc結構域、用於PD-L1之結合結構域、及用於41-BB之結合結構域。在一個實施例中,GNC蛋白質可包括具有與SEQ ID NO. 80及82之一同源性百分比的胺基酸序列。同源性百分比不小於70%、80%、90%、95%、98%或99%。在一個實施例中,GNC蛋白質為四特異性抗體。
在一個實施例中,引導及導航控制(GNC)蛋白質具有N末端及C末端,其自N末端至C末端串聯地包含用於4-1BB之結合結構域、用於PD-L1之結合結構域、IgG Fc結構域、用於ROR1之結合結構域、及用於CD3之結合結構域。在一個實施例中,GNC蛋白質包括具有與SEQ ID NO. 88及90之一同源性百分比的胺基酸序列。同源性百分比不小於70%、80%、90%、95%、98%或99%。在一個實施例中,GNC蛋白質為四特異性抗體。
引導及導航控制(GNC)蛋白質具有N末端及C末端,其自N末端至C末端串聯地包含用於CD3之結合結構域、用於CD19之結合結構域、IgG Fc結構域、用於PD-L1之結合結構域、及用於4-1BB之結合結構域。在一個實施例中,GNC蛋白質包括具有與SEQ ID NO. 104及106之一同源性百分比的胺基酸序列。同源性百分比不小於70%、80%、90%、95%、98%或99%。在一個實施例中,GNC蛋白質為四特異性抗體。
在一個實施例中,GNC蛋白質包含具有與SEQ ID NO. 50、52、80、82、84、86、88、90、92、94、96、98、100、102、104、106、108及110之一同源性百分比的胺基酸,且同源性百分比不小於70%、80%、90%、95%、98%或99%。
在另一態樣中,本申請案提供編碼本案揭示的GNC蛋白質或其片段之核酸序列。在一個實施例中,核酸具有與SEQ ID NO. 49、51、79、81、83、85、87、89、91、93、95、97、99、101、103、105、107、及109之一同源性百分比,且同源性百分比不小於70%、80%、90%、95%、98%或99%。
在一個實施例中,引導及導航控制(GNC)蛋白質包含細胞毒性細胞結合部分及癌症靶向部分。任何細胞毒性細胞可為所揭示GNC蛋白質之潛在結合標靶。細胞毒性細胞之實例包括而不限於T細胞、NK細胞、巨噬細胞、及樹狀細胞。
在一個實施例中,GNC蛋白質包括T細胞結合部分。T細胞結合部分具有對T細胞受體之結合特異性。示例性T細胞受體包括而不限於CD3、CD28、PDL1、PD1、OX40、4-1BB、GITR、TIGIT、TIM-3、LAG-3、CTLA4、CD40L、VISTA、ICOS、BTLA、Light、CD30、NKp30、CD28H、CD27、CD226、CD96、CD112R、A2AR、CD160、CD244、CECAM1、CD200R、TNFRSF25 (DR3)、或其組合。
在一個實施例中,GNC蛋白質包括NK細胞結合部分。NK細胞結合部分具有對NK細胞受體之結合特異性。示例性NK細胞受體包括而不限於用於活化NK細胞之受體,諸如CD16、NKG2D、KIR2DS1、KIR2DS2、KIR2DS4、KIR3DS1、NKG2C、NKG2E、NKG2H;促效劑受體,諸如NKp30a、NKp30b、NKp46、NKp80、DNAM-1、CD96、CD160、4-1BB、GITR、CD27、OX-40、CRTAM;及拮抗劑受體,諸如KIR2DL1、KIR2DL2、KIR2DL3、KIR3DL1、KIR3DL2、KIR3DL3、NKG2A、NKp30c、TIGIT、SIGLEC7、SIGLEC9、LILR、LAIR-1、KLRG1、PD-1、CTLA-4、CD161。
在一個實施例中,GNC蛋白質包括巨噬細胞結合部分。巨噬細胞結合部分具有對巨噬細胞受體之結合特異性。示例性巨噬細胞受體包括而不限於巨噬細胞上之促效劑受體,諸如TLR2、TLR4、CD16、CD64、CD40、CD80、CD86、TREM-1、TREM-2、ILT-1、ILT-6a、ILT-7、ILT-8、EMR2、Dectin-1、CD69;及拮抗劑受體,諸如CD32b、SIRPα、LAIR-1、VISTA、TIM-3、CD200R、CD300a、CD300f、SIGLEC1、SIGLEC3、SIGLEC5、SIGLEC7、SIGLEC9、ILT-2、ILT-3、ILT-4、ILT-5、LILRB3、LILRB4、DCIR;及其他表面受體,諸如CSF-1R、LOX-1、CCR2、FRβ、CD163、CR3、DC-SIGN、CD206、SR-A、CD36、MARCO。
在一個實施例中,GNC蛋白質包括樹狀細胞結合部分。樹狀細胞結合部分具有對樹狀細胞受體之結合特異性。示例性樹狀細胞受體包括而不限於樹狀細胞上之促效劑受體,諸如TLR、CD16、CD64、CD40、CD80、CD86、HVEM、CD70;拮抗劑受體,諸如VISTA、TIM-3、LAG-3、BTLA;及其他表面受體,諸如CSF-1R、LOX-1、CCR7、DC-SIGN、GM-CSF-R、IL-4R、IL-10R、CD36、CD206、DCIR、RIG-1、CLEC9A、CXCR4。
癌症靶向部分具有對癌細胞受體之結合特異性。示例性癌細胞受體包括而不限於BCMA、CD19、CD20、CD33、CD123、CD22、CD30、ROR1、CEA、HER2、EGFR、EGFRvIII、LMP1、LMP2A、間皮素、PSMA、EpCAM、磷脂醯肌醇蛋白聚糖-3、gpA33、GD2、TROP2、或其組合。
在一個實施例中,GNC蛋白質包含至少一個T細胞結合部分及至少一個癌細胞結合部分,其中該T細胞結合部分對T細胞受體具有結合特異性,該T細胞受體包含CD3、CD28、PDL1、PD1、OX40、4-1BB、GITR、TIGIT、TIM-3、LAG-3、CTLA4、CD40、VISTA、ICOS、BTLA、Light、CD30、CD27、或其組合,且其中該癌細胞結合部分對癌細胞受體具有結合特異性。
在一個實施例中,GNC蛋白質能夠藉由將T細胞結合部分結合至T細胞上之T細胞受體而活化T細胞。在一個實施例中,GNC蛋白質包含雙特異性抗體或抗體單體、三特異性抗體或抗體單體、四特異性抗體或抗體單體、其抗原結合片段、或其組合。
在一個實施例中,GNC蛋白質可具有第一部分及第二部分。在一個實施例中,第一部分可包括T細胞結合部分、NK細胞結合部分、巨噬細胞結合部分、或樹狀細胞結合部分。第二部分包含癌症靶向部分。
本申請案進一步提供併入有本文所揭示的GNC蛋白質之細胞毒性細胞。在一個實施例中,細胞毒性細胞包括GNC蛋白質及細胞毒性細胞。細胞毒性細胞可為T細胞、NK細胞、巨噬細胞、樹狀細胞、或其組合。在一個實施例中,T細胞可為自體T細胞、異體T細胞、或全適供應者T細胞(universal donor T cell)。在一個實施例中,細胞毒性細胞包括具有T細胞活化受體及T細胞共刺激受體之T細胞,及經由與T細胞活化受體、T細胞共刺激受體、或其組合的相互作用結合至T細胞之GNC蛋白質。
本申請案進一步提供併入有本文所揭示的GNC蛋白質之癌細胞。在一個實施例中,癌細胞包含具有腫瘤相關聯抗原之癌細胞,及經由與腫瘤相關聯抗原相互作用結合至癌細胞的如請求項1所述之GNC蛋白質。
本申請案進一步提供併入有本文所揭示的GNC蛋白質之生物學複合物。在一個實施例中,生物學複合物包括具有T細胞活化受體及T細胞共刺激受體之T細胞、具有腫瘤相關聯抗原之癌細胞、及如請求項1所述之GNC蛋白質,其中GNC蛋白質經由與T細胞活化受體、T細胞共刺激受體、或其組合相互作用結合至T細胞,且其中GNC蛋白質經由與腫瘤相關聯抗原相互作用結合至癌細胞。
在另一態樣中,本申請案提供適用於治療癌症病狀的醫藥組成物。在一個實施例中,醫藥組成物包括本文所揭示的GNC蛋白質或細胞毒性細胞,及醫藥學上可接受的載劑。
在另一態樣中,本申請案提供用於製備及使用所揭示GNC蛋白質之方法。
在另一態樣中,本申請案提供用於治療患有癌症之受試者的方法。在一個實施例中,方法包括向受試者投與有效量之本文揭示的醫藥組合物的步驟。
本申請案之目標及優點將自以下本申請案之較佳實施例之詳細說明結合隨附圖式而變得明顯。
在以下詳細說明中,參考隨附圖式,其形成本文之一部分。在圖式中,除非上下文另外指定,否則類似符號典型地標示類似的組分。描述在詳細說明、圖式、及申請專利範圍中的說明性實施例不意謂為限制性的。可使用其他實施例,且可做出其他改變,而不脫離本文提出的標的之精神或範疇。易於理解的是,如本文大體上描述及圖式中說明的本揭示內容之態樣可以多種不同的配置來佈置、替代、組合、分離、及設計,其全部明確地涵蓋在本文中。
本申請案係關於製備及使用GNC蛋白質之方法。在一個實施例中,引導導航控制(GNC)蛋白質可包括多個抗原特異性結合結構域(AgBD)且可具有經由結合T細胞及腫瘤細胞上之多個表面分子而導引T細胞(或其他效應細胞)至癌細胞(或其他靶細胞)之能力(第1圖)。在一個實施例中,GNC蛋白質可由部分1及部分2構成,該部分1用於結合T細胞上之至少一個表面分子,且該部分2用於結合癌細胞上之至少一個表面抗原(表1A)。
在T細胞療法中,細胞毒性T細胞係藉由T細胞增殖傳訊,以及經由其表面上之促效劑受體或拮抗劑受體的共刺激傳訊來調節。為調節該些傳訊,以及T細胞與癌症之間的相互作用,多個AgBD可分別及獨立地包括來用於部分1及部分2。GNC蛋白質可具有至少一個連接子以連接部分1及部分2。連接子之長度可變化。在一個實施例中,連接子可為共價鍵。在一個實施例中,連接子可為具有約1至約100個胺基酸殘基之肽。
在一些實施例中,任何連接子分子可用於在活體外或活體內藉由使用DNA/RNA之互補連接子或蛋白質-蛋白質相互作用將兩個或更多個AgBD連接在一起,該等相互作用包括但不限於生物素-抗生物素蛋白、白胺酸-拉鍊、及任何兩個雜合體陽性蛋白質之彼相互作用。
在一些實施例中,連接子可為抗體主鏈結構或抗體片段,使得GNC蛋白質及GNC抗體可具有相同的含義,如第2圖所示,示例性四特異性GNC抗體結構。在一個實施例中,該GNC蛋白質可為雙特異性、三特異性、四特異性、五特異性、六特異性、七特異性、或八特異性蛋白質。在一個實施例中,GNC蛋白質可為單株抗體。在一個實施例中,該GNC蛋白質可為雙特異性、三特異性、四特異性、五特異性、六特異性、七特異性、或八特異性抗體單體。在一個實施例中,該GNC蛋白質可為雙特異性、三特異性、四特異性、五特異性、六特異性、七特異性、或八特異性抗體單體。
GNC蛋白質或抗體可能能夠導引活體內或離體的藉由多個AgBD介導的T細胞對癌細胞之結合(第3圖)。T細胞可來源於相同患者或不同個體,且癌細胞可在活體內、活體外、或離體存在。提供於本申請案中之實例賦能GNC蛋白質作為T細胞療法,亦即,GNC-T療法中之啟始劑(prime agent)以用於在授受性轉移之前離體活化及控制細胞毒性T細胞。
除T細胞之外,其他細胞毒性細胞可藉由GNC蛋白質用於癌症殺死或預防目的。表1B展示具有NK細胞結合結構域之GNC蛋白質中的功能部分(部分1及部分2)及抗原結合結構域之示例性組合物。表1C展示具有巨噬細胞結合結構域之GNC蛋白質中的功能部分(部分1及部分2)及抗原結合結構域之示例性組合物。表1D展示具有樹狀細胞結合結構域之GNC蛋白質中的功能部分(部分1及部分2)及抗原結合結構域之示例性組合物。
多個AgBD可歸因於其分別與諸如T細胞及癌細胞之細胞毒性細胞的界面而分為部分1及部分2 (表1A)。然而,多個AgBD之重排可為隨機的且以不等數量重排(表2)。具有兩個AgBD之GNC蛋白質可同時結合至諸如T細胞上之CD3的表面分子及諸如腫瘤細胞上之ROR1的腫瘤抗原,以用於將T細胞重導引或引導至腫瘤細胞。例如特異地結合至41BB的第三AgBD之添加可幫助增強抗CD3誘導之T細胞活化,因為41BB為共刺激因子且結合刺激其對活化T細胞之促效劑活性。例如特異地結合至腫瘤細胞上之PD-L1的第四AgBD向GNC蛋白質之添加可阻斷腫瘤細胞上的PD-L1之抑制路徑,該抑制路徑係經由其結合至T細胞上之PD-1介導。利用該些基本原理,GNC蛋白質可經設計及構造來獲得多個AgBD以特定地用於結合不等數量之T細胞拮抗劑及促效劑,不僅將活化T細胞重導引至腫瘤細胞而且控制其活體內活性(表2)。因此,GNC蛋白質之設計可為任何多特異性蛋白質。表3提供具有抗體結合結構域之特異性的一些示例性GNC蛋白質及抗體。
在一個實施例中,GNC蛋白質可包括藉由兩個官能基團表徵的多特異性抗原結合部分:部分1包含多個抗原結合結構域(AgBD),其特異性涉及T細胞活化、促效劑共刺激、及/或抑制拮抗劑活性;且部分2包含至少一種癌細胞結合特異性。GNC蛋白質可同時結合至諸如T細胞之CD3的表面分子及諸如腫瘤細胞之ROR1的腫瘤抗原,進而將T細胞之重導引或引導至腫瘤細胞。在GNC蛋白質中添加第三結合結構域可經由其直接結合41BB而幫助增強CD3-誘導之T細胞活化,該41BB為發揮促效劑活性之共刺激因子。此外,第四結合結構域在GNC蛋白質中之添加可結合至腫瘤細胞上之PD-L1以阻斷腫瘤細胞上的PD-L1之抑制路徑,該抑制路徑係經由其結合至T細胞上之PD-1介導。在一些實施例中,GNC蛋白質獲得對重導引活化T細胞至腫瘤細胞之多重結合能力,且多重結合可幫助經由調節促效劑或拮抗劑活性或兩者來調節T細胞活化。一些結合能力可類似於CAR-T細胞上之嵌合抗原受體或雙特異性抗體、諸如BiTe抗體之彼能力。在不欲藉由理論約束的情況下,經由各種結構域與細胞毒性細胞受體及腫瘤相關聯抗原之相互作用,GNC蛋白質可提供作為治療劑的相比於傳統的基於細胞之治療劑(諸如CAR-T及抗體療法)的顯著優點,包括而不限於改良的結合功效、最佳化的細胞傳訊及細胞毒性,以及減少的副作用,諸如減少的細胞介素風暴症候群之嚴重程度。
在一個實施例中,本申請案提供具有4個不同結合結構域的示例性GNC蛋白質。GNC蛋白質可為「四特異性抗體」,其中其連接子及主鏈包含抗體片段。在4個不同的抗原結合結構域中,一個抗原結合結構域特異地結合至T細胞上之CD3,第二結合結構域針對腫瘤相關聯抗原為特異性的,該腫瘤相關聯抗原包括但不限於其他腫瘤抗原,諸如ROR1、CEA、HER2、EGFR、EGFRvIII、LMP1、LMP2A、間皮素、PSMA、EpCAM、磷脂醯肌醇蛋白聚糖-3、gpA33、GD2、TROP2、BCMA、CD19、CD20、CD33、CD123、CD22、CD30,且第三結合結構域及第四結合結構域針對兩個相異免疫查核點調節劑為特異性的,該等相異免疫查核點調節劑即為PD-L1、PD-1、OX40、4-1BB、GITR、TIGIT、TIM-3、LAG-3、CTLA4、CD40、VISTA、ICOS、BTLA、Light、HVEM、CD73、CD39等等。由於其在組合物中的功能及變化方面定義,GNC蛋白質可分類為新類別的用於治療癌症的免疫調節劑。表4展示示例性四特異性GNC抗體之列表。
在一個實施例中,GNC介導之免疫療法可包括抗體療法及細胞療法之類型。本文中,優點可包括但不限於,IgG Fc結構域之包括可相較於雙特異性BiTe分子賦予在血清中較長半衰期之特性;其次,對免疫查核點調節劑為特異性的兩個結合結構域之包括可抑制抑制路徑且同時接合共刺激路徑;第三,將T細胞上之CD3與腫瘤相關聯抗原交聯重導引及引導T細胞來殺死腫瘤細胞而無需自患者移除T細胞且將其遺傳修飾為對腫瘤細胞為特異性的,之後將其重引入回患者中,此亦稱為嵌合抗原受體T細胞(CAR-T)療法;且第四,GNC蛋白質介導的抗體療法或T細胞療法不涉及T細胞之遺傳修飾,後者可帶來轉變經修飾T細胞至純系擴張,亦即,T細胞白血病之風險。
利用結合能力之一或多個添加,GNC蛋白質介導之免疫療法優於習知免疫療法之優點包括但不限於,首先,IgG Fc結構域之包括可相較於雙特異性BiTe分子賦予在血清中較長半衰期之特性;其次,對免疫查核點調節劑為特異性的兩個結合結構域之包括可抑制抑制路徑且同時接合共刺激路徑;第三,將T細胞上之CD3與腫瘤相關聯抗原交聯重導引及引導T細胞來殺死腫瘤細胞而無需自患者移除T細胞且將其遺傳修飾為對腫瘤細胞為特異性的,之後將其重引入回患者中,此亦稱為嵌合抗原受體T細胞(CAR-T)療法;且第四,GNC蛋白質介導的抗體療法或T細胞療法不涉及T細胞之遺傳修飾,後者可帶來轉變經修飾T細胞至純系擴張,亦即,T細胞白血病之風險。
本揭示內容可參考本文包括的特定實施例及實例之以下詳細說明得以更容易地理解。儘管本揭示內容已參考其某些實施例之特定細節來描述,但此等細節不意欲視為對本揭示內容之範疇的限制。
實例
儘管以下實例係僅以說明方式而非限制方式來提供。熟習此項技術者將容易地認識到可改變或修改來產生基本上相同或類似結果的各種非臨界參數。
實例
1
:四特異性
GNC
抗體結合至經人類
ROR1
轉染之
CHO
細胞的
FACS
分析
針對結合至穩定地表現全長人類ROR1之中國倉鼠卵巢(Chinese hamster ovary;CHO)細胞來測試表3及4中列出的四特異性GNC抗體。以2X最終濃度製備抗體且在50 ul之PBS/2% FBS中跨於96孔板之3個孔滴定1:5,且隨後添加於50 ul PBS/2%FBS中之5,000個ROR1-CHO細胞。將此混合物在冰上培養30分鐘,利用200 ul PBS/2%FBS洗滌一次,且隨後以儲備液之1:1000稀釋來添加二級抗體 PE山羊抗人類IgG Fc,且將此混合物在冰上培養30分鐘。將細胞洗滌2 x 200 ul PBS/2%FBS,再懸浮於50 ul PBS/2%FBS中且在BD LSRFORTESSA上分析且在第4圖中展示結合分佈。具有對ROR1之Ig結構域為特異性的323H7結合結構域的四特異性抗體SI-35E18、19、及20比具有對ROR1之捲曲蛋白結構域為特異性的338H4結合結構域的四特異性GNC抗體SI-3521、22、及23展示更高的結合,且具有對ROR1之kringle結構域為特異性的330F11結合結構域的四特異性GNC抗體SI-3524、25、及26不結合。
實例
2
:四特異性
GNC
抗體結合至經人類
41BB
轉染之
CHO
細胞的
FACS
分析
針對結合至穩定地表現全長人類ROR1之中國倉鼠卵巢(CHO)細胞來測試表3及4中列出的四特異性GNC抗體。以2X最終濃度製備抗體且在50 ul之PBS/2% FBS中跨於96孔板之3個孔滴定1:5,且隨後添加於50 ul PBS/2%FBS中之5,000個ROR1-CHO細胞。將此混合物在冰上培養30分鐘,利用200 ul PBS/2%FBS洗滌一次,且隨後以儲備液之1:1000稀釋來添加二級抗體PE山羊抗人類IgG Fc,且將此混合物在冰上培養30分鐘。將細胞洗滌2 x 200 ul PBS/2%FBS,再懸浮於50 ul PBS/2%FBS中,且在BD LSRFORTESSA上分析並在第5圖中展示結合分佈。除對照SI-27E12外,所有四特異性GNC抗體含有41BB結合結構域460C3、420H5、或466F6,且以變化的強度結合至表現41BB之CHO細胞。
實例
3
:四特異性
GNC
抗體結合至經人類
PDL1
轉染之
CHO
細胞的
FACS
分析
針對結合至穩定地表現全長人類ROR1之中國倉鼠卵巢細胞(CHO)細胞測試表3及4中列出的四特異性GNC抗體。以2X最終濃度製備抗體且在50 ul之PBS/2% FBS中跨於96孔板之3個孔滴定1:5,且隨後添加於50 ul PBS/2%FBS中之5,000個ROR1-CHO細胞。將此混合物在冰上培養30分鐘,利用200 ul PBS/2%FBS洗滌一次,且隨後以儲備液之1:1000稀釋來添加二級抗體PE山羊抗人類IgG Fc,且將此混合物在冰上培養30分鐘。將細胞洗滌2 x 200 ul PBS/2%FBS,再懸浮於50 ul PBS/2%FBS中且在BD LSRFORTESSA上分析且在第6圖中展示結合分佈。除對照SI-27E15外,所有四特異性GNC抗體含有相同的PDL1結合結構域PL230C6,且展示對表現PDL1之CHO細胞的極類似的結合強度。
實例4:以末梢血液單核細胞作為效應物且以B-急性淋巴母細胞性白血病(B-Acute Lymphoblastic Leukemia;B-ALL)細胞系Kasumi-2作為標靶的重導引T細胞細胞毒性(Re-directed T cell cytotoxicity;RTCC)分析
就針對B-ALL細胞系Kasumi 2之RTCC活性,使用人類末梢血液單核細胞(peripheral blood mononuclear cell;PBMC)作為效應物來測試表3及4中列出的四特異性GNC抗體。在37℃下,在10 ml之培養基中將Kasumi 2靶細胞5 x 106
用CFSE (Invitrogen, #C34554)以0.5 uM標記20分鐘。將細胞用50 ml之培養基洗滌3次,之後再懸浮於10 ml中,隨後再次計數。以2X最終濃度製備抗體且在200 ul之RPMI + 10%FBS中跨於96孔板之10個孔滴定1:3。藉由標準ficoll密度梯度自「leukopak」純化人類PBMC,該leukopak為自正常人類末梢血液收集的富集白細胞除去法(leukapheresis)產物。在最終目標96孔板中,藉由將100 ul之靶細胞(5,000)、50 ul之PBMC (25,000)、及100 ul之每一抗體稀釋液添加至分析之每一孔來組合靶細胞、PBMC、及連續滴定抗體。在37℃下將分析板培養大致72小時且隨後收穫每一分析孔之內容物且分析所剩餘的CFSE標記靶細胞之數量。如第7圖上所示,四特異性GNC抗體全部含有相同的PDL1結合結構域PL230C6、相同的ROR1結合結構域323H7、及相同的CD3結合結構域284A10,但是具有41BB結合結構域460C3、420H5、及466F6之一,且除不具有41BB結合結構域之對照SI-27E12外,展示相較於對照的較大RTCC活性,但似乎在四特異性GNC抗體SI-35E18、19、及20處為類似有效的。
實例5:以CD8+、CD45RO+記憶T細胞作為效應物且以B-急性淋巴母細胞性白血病(B-Acute Lymphoblastic Leukemia;B-ALL)細胞系Kasumi-2作為標靶的重導引T細胞細胞毒性(Re-directed T cell cytotoxicity;RTCC)分析
就針對B-ALL細胞系Kasumi 2之RTCC活性,使用人類CD8+、CD45RO+記憶T細胞作為效應物來測試表3及4中列出的四特異性GNC抗體。在37℃下,在10 ml之培養基中將Kasumi 2靶細胞5 x 106
用CFSE (Invitrogen, #C34554)以0.5 uM標記20分鐘。將細胞用50 ml之培養基洗滌3次,之後再懸浮於10 ml中,隨後再次計數。以2X最終濃度製備抗體且在200 ul之RPMI + 10%FBS中跨於96孔板之10個孔滴定1:3。按照製造商之協定,使用EasySep™人類記憶CD8+ T細胞富集套組(Stemcell Technologies, #19159)自來自正常供體之PBMC富集人類CD8+、CD45RO+記憶T細胞。最終細胞群體係藉由FACS分析測定為98% CD8+、CD45RO+ T細胞。在最終目標96孔板中,藉由將100 ul之靶細胞(5,000)、50 ul之CD8+、CD45RO+記憶T細胞(25,000)、及100 ul之每一抗體稀釋液添加至分析之每一孔來組合靶細胞、T細胞、及連續滴定抗體。在37℃下將分析板培養大致72小時且隨後收穫每一分析孔之內容物且分析所剩餘的CFSE標記靶細胞之數量。如第8圖上所示,四特異性抗體全部含有相同的PDL1結合結構域PL230C6、相同的ROR1結合結構域323H7、及相同的CD3結合結構域284A10,但具有41BB結合結構域460C3、420H5、及466F6之一,且相較於不含有41BB、PDL1、ROR1、或CD3結合結構域之一的對照展示較大的RTCC活性。
實例6:以CD8+、CD45RA+初始T細胞作為效應物且以B-急性淋巴母細胞性白血病(B-Acute Lymphoblastic Leukemia;B-ALL)細胞系Kasumi-2作為標靶的重導引T細胞細胞毒性(Re-directed T cell cytotoxicity;RTCC)分析
就針對B-ALL細胞系Kasumi 2之RTCC活性,使用人類CD8+、CD45RA+記憶T細胞作為效應物來測試表3及4中列出的四特異性特異抗體。在37℃下,在10 ml之培養基中將Kasumi 2靶細胞5 x 10e6用CFSE (Invitrogen, #C34554)以0.5 uM標記20分鐘。將細胞用50 ml之培養基洗滌3次,之後再懸浮於10 ml中,隨後再次計數。以2X最終濃度製備抗體且在200 ul之RPMI + 10%FBS中跨於96孔板之10個孔滴定1:3。按照製造商之協定,使用EasySep™人類初始CD8+ T細胞分離套組(Stemcell Technologies, #19258)自來自正常供體之末梢血液單核細胞富集人類CD8+、CD45RA+記憶T細胞。最終細胞群體係藉由FACS分析測定為98% CD8+、CD45RA+ T細胞(資料未展示)。在最終目標96孔板中,藉由將100 ul之靶細胞(5,000)、50 ul之CD8+、CD45RO+ T細胞(25,000)、及100 ul之每一抗體稀釋液添加至分析之每一孔來組合靶細胞、T細胞、及連續滴定抗體。在37℃下將分析板培養大致72小時且隨後收穫每一分析孔之內容物且分析所剩餘的CFSE標記靶細胞之數量。如第9圖上所示,所有四特異性抗體含有相同的PDL1結合結構域PL230C6、相同的ROR1結合結構域323H7、及相同的CD3結合結構域284A10,但具有41BB結合結構域460C3、420H5、及466F6之一,且相較於不含有41BB、PDL1、ROR1、或CD3結合結構域之一的對照展示較大的RTCC活性。
實例7:以末梢血液單核細胞作為效應物且以B-急性淋巴母細胞性白血病(B-Acute Lymphoblastic Leukemia;B-ALL)細胞系Kasumi-2作為標靶的重導引T細胞細胞毒性(Re-directed T cell cytotoxicity;RTCC)分析
就針對B-ALL細胞系Kasumi 2之RTCC活性,使用人類末梢血液單核細胞(peripheral blood mononuclear cell;PBMC)作為效應物來測試表3及4中列出的四特異性特異抗體。在37℃下,在10 ml之培養基中將Kasumi 2靶細胞5 x 106
用CFSE (Invitrogen, #C34554)以0.5 uM標記20分鐘。將細胞用50 ml之培養基洗滌3次,之後再懸浮於10 ml中,隨後再次計數。以2X最終濃度製備抗體且在200 ul之RPMI + 10%FBS中跨於96孔板之10個孔滴定1:3。藉由標準ficoll密度梯度自「leukopak」純化人類PBMC,該leukopak為自正常人類末梢血液收集的富集白細胞除去法(leukapheresis)產物。在最終目標96孔板中,藉由將100 ul之靶細胞(5,000)、50 ul之PBMC (25,000)、及100 ul之每一抗體稀釋液添加至分析之每一孔來組合靶細胞、PBMC、及連續滴定抗體。在37℃下將分析板培養大致72小時且隨後收穫每一分析孔之內容物且分析所剩餘的CFSE標記靶細胞之數量。如第10圖上所示,四特異性GNC抗體全部含有相同的PDL1結合結構域PL230C6、相同的ROR1結合結構域338H4、及相同的CD3結合結構域284A10,但是具有41BB結合結構域460C3、420H5、及466F6之一,且除不具有41BB結合結構域之對照SI-35E36外,展示相較於對照的較大RTCC活性,但似乎在四特異性GNC抗體SI-35E18、19、及20處為類似有效的。
實例8:以CD8+、CD45RO+記憶T細胞作為效應物且以B-急性淋巴母細胞性白血病(B-Acute Lymphoblastic Leukemia;B-ALL)細胞系Kasumi-2作為標靶的重導引T細胞細胞毒性(Re-directed T cell cytotoxicity;RTCC)分析
就針對B-ALL細胞系Kasumi 2之RTCC活性,使用人類CD8+、CD45RO+記憶T細胞作為效應物來測試表3及4中列出的四特異性GNC抗體。在37℃下,在10 ml之培養基中將Kasumi 2靶細胞5 x 106
用CFSE (Invitrogen, #C34554)以0.5 uM標記20分鐘。將細胞用50 ml之培養基洗滌3次,之後再懸浮於10 ml中,隨後再次計數。以2X最終濃度製備抗體且在200 ul之RPMI + 10%FBS中跨於96孔板之10個孔滴定1:3。按照製造商之協定,使用EasySep™人類記憶CD8+ T細胞富集套組(Stemcell Technologies, #19159)自來自正常供體之PBMC富集人類CD8+、CD45RO+記憶T細胞。最終細胞群體係藉由FACS分析測定為98% CD8+、CD45RO+ T細胞(資料未展示)。在最終目標96孔板中,藉由將100 ul之靶細胞(5,000)、50 ul之CD8+、CD45RO+記憶T細胞(25,000)、及100 ul之每一抗體稀釋液添加至分析之每一孔來組合靶細胞、T細胞、及連續滴定抗體。在37℃下將分析板培養大致72小時且隨後收穫每一分析孔之內容物且分析所剩餘的CFSE標記靶細胞之數量。如第11圖上所示,四特異性GNC抗體全部含有相同的PDL1結合結構域PL230C6、相同的ROR1結合結構域338H4、及相同的CD3結合結構域284A10,但具有41BB結合結構域460C3、420H5、及466F6之一,且相較於不含有41BB、PDL1、ROR1、或CD3結合結構域之一的對照展示較大的RTCC活性。
實例9:以CD8+、CD45RA+初始T細胞作為效應物且以B-急性淋巴母細胞性白血病(B-Acute Lymphoblastic Leukemia;B-ALL)細胞系Kasumi-2作為標靶的重導引T細胞細胞毒性(Re-directed T cell cytotoxicity;RTCC)分析
就針對B-ALL細胞系Kasumi 2之RTCC活性,使用人類CD8+、CD45RA+記憶T細胞作為效應物來測試表3及4中列出的四特異性GNC抗體。在37℃下,在10 ml之培養基中將Kasumi 2靶細胞5 x 106用CFSE (Invitrogen, #C34554)以0.5 uM標記20分鐘。將細胞用50 ml之培養基洗滌3次,之後再懸浮於10 ml中,隨後再次計數。以2X最終濃度製備抗體且在200 ul之RPMI + 10%FBS中跨於96孔板之10個孔滴定1:3。按照製造商之協定,使用EasySep™人類初始CD8+ T細胞分離套組(Stemcell Technologies, #19258)自來自正常供體之PBMC富集人類CD8+、CD45RA+記憶T細胞。最終細胞群體係藉由FACS分析測定為98% CD8+、CD45RA+ T細胞。在最終目標96孔板中,藉由將100 ul之靶細胞(5,000)、50 ul之CD8+、CD45RO+ T細胞(25,000)、及100 ul之每一抗體稀釋液添加至分析之每一孔來組合靶細胞、T細胞、及連續滴定抗體。在37℃下將分析板培養大致72小時且隨後收穫每一分析孔之內容物且分析所剩餘的CFSE標記靶細胞之數量。如第12圖上所示,四特異性GNC抗體全部含有相同的PDL1結合結構域PL230C6、相同的ROR1結合結構域338H4、及相同的CD3結合結構域284A10,但具有41BB結合結構域460C3、420H5、及466F6之一,但相較於不含有41BB、PDL1、ROR1、或CD3結合結構域之一的對照不展示較大的RTCC活性。
實例
10
:針對膀胱癌細胞系
UM-UC-3-EGFRvIII
的重導引
panT
細胞細胞毒性。
評定表5中列出的一組四特異性GNC抗體的溶解靶細胞UM-UC-3-EGFRvIII之能力。用EasySep™人類Pan T細胞分離套組(Stemcell Technologies)分離PanT細胞。UM-UC-3-EGFRvIII細胞系穩定地表現經由慢病毒轉導(Sartorius)遞送的核局部化紅色螢光蛋白(Red Fluorescent Protein;RFP)。將UM-UC-3-EGFRvIII-RFP腫瘤細胞用panT細胞共培養。經由計數在用panT細胞進行96 h共培養之後留在培養物中的活標靶之數量,用流式細胞術(BD LSRFortessa)評定靶細胞溶解。兩種四特異性抗體SI-39E18及SI-39E29為在靶腫瘤細胞溶解方面最有效的(第13圖)。該兩種分子亦由用於CD3及腫瘤抗原之相鄰結合結構域構成(表5)。
實例
11
:回應於用靶向
EGFRvIII
之四特異性抗體進行的治療之
CD8 T
細胞增殖。
評定表5中列出的一組四特異性GNC抗體的在靶細胞UM-UC-3-EGFRvIII存在下刺激CD8 T細胞增殖之能力。用CellTrace紫色染料(Thermo Fisher Scientific)標記PanT細胞。將UM-UC-3-EGFRvIII-RFP腫瘤細胞用panT細胞共培養。經由在96 h共培養之後稀釋CellTrace紫色染料,用流式細胞術(BD LSRFortessa)評定CD8 T細胞增殖。兩種四特異性GNC抗體SI-39E18及SI-39E29為在靶細胞存在下刺激CD8 T細胞增殖方面最有效的(第14圖)。該兩種分子由用於CD3及腫瘤抗原之相鄰結合結構域構成(表5)。具有強T細胞刺激活性之其他分子包括含有相鄰CD3及PD-L1結構域之結構(表5)。
實例
12
:回應於用靶向
EGFRvIII
之四特異性抗體進行的治療之
IFNγ
分泌。
評定表5中列出的一組四特異性GNC抗體的藉由PBMC誘導IFNγ分泌之能力。藉由Ficoll梯度分離PBMC。用測試分子培養PBMC 96 h。收集上清液且使用ELISA (R & D Systems)分析IFNγ之存在(第15圖)。在此項研究中具有最強活性的四特異性GNC抗體全部含有相鄰CD3及PD-L1結構域(表5)。活性最小群組具有帶相鄰CD3及腫瘤抗原或4-1BB結構域之分子。對該組四特異性GNC抗體的唯一例外為SI-39E18,其含有相鄰CD3及腫瘤抗原結構域。此分子刺激IFNγ之適度產生,其小於具有相鄰CD3及PD-L1結構域之分子的活性最大群組,但大於具有類似結構佈置之其他分子。IFNγ之適度產生可能對此藥劑之抗腫瘤活性為有益的。
實例
13
:針對膀胱癌細胞系
UM-UC-3-EGFRvIII
之重導引初始
T
細胞細胞毒性。
測試四特異性GNC抗體SI-39E18的重導引初始T細胞以溶解靶細胞UM-UC-3-EGFRvIII之能力。用EasySep™人類初始PanT細胞分離套組(Stemcell Technologies)分離初始T細胞。將UM-UC-3-EGFRvIII-RFP腫瘤細胞用初始或panT細胞共培養。藉由計數RFP標記的腫瘤細胞核來評定腫瘤細胞之溶解。在肝細胞成像器IncuCyte上獲取影像(Sartorius)。在120小時之培養之後評定抗體之活性。在較低效應物對標靶比率2.5至1下測試治療。SI-39E18在重導引初始T細胞方面為有效的。對初始T細胞而言EC50為22.08 pM,對panT細胞而言,EC50為0.07 pM (第16圖)。
實例
14
:
PBMC
對用靶向
EGFRvIII
之四特異性
GNC
抗體進行的治療、
CD8 T
細胞之增殖的反應。
評定表1中列出一組四特異性GNC抗體在不存在靶細胞的情況下誘導CD8 T細胞增殖之能力。用CellTrace紫色染料(Thermo Fisher Scientific)標記PBMC且用測試分子培養96 h。經由稀釋CellTrace紫色染料,用流式細胞術(BD LSRFortessa)評定CD8 T細胞增殖。在此項研究中最有效的分子共有結構相似性(第17圖)。所有該些分子含有相鄰CD3及PD-L1結構域(表5)。
實例
15.
在單核細胞存在下針對膀胱癌細胞系
UM-UC-3-EGFRvIII
之重導引
panT
細胞活性。
評定表5中列出的一組四特異性GNC抗體的在單核細胞存在下溶解靶細胞UM-UC-3-EGFRvIII之能力。用EasySep™人類單核細胞分離套組(Stemcell Technologies)自PBMC分離單核細胞。將UM-UC-3-EGFRvIII-RFP腫瘤細胞用panT細胞及單核細胞共培養。藉由計數RFP標記的腫瘤細胞核來評定靶細胞溶解。在肝細胞成像器IncuCyte上獲取影像(Sartorius)。在96小時之培養之後評定抗體之活性。兩種四特異性GNC抗體SI-39E18及SI-39E29連同含有相鄰CD3及PD-L1結合結構域之分子(表5)在靶腫瘤細胞溶解方面為最有效的(第18圖)。
實例
16.
針對膀胱癌細胞系
UM-UC-3-EGFRvIII
之重導引
PBMC
細胞毒性、不同
4-1BB
結構域之功能活性及
PD-L1
及
4-1BB
結構域之功能影響。
評定表5中列出的四特異性GNC抗體的重導引PBMC癌細胞系UM-UC-3-EGFRvIII (UM-UC-3-EGFRvIII)之能力。將UM-UC-3-EGFRvIII-RFP腫瘤細胞用PBMC共培養。藉由計數RFP標記的腫瘤細胞核來評定腫瘤細胞之溶解。在肝細胞成像器IncuCyte上獲取影像(Sartorius)。在96小時之培養之後評定抗體之活性。具有不同4-1BB結構域之四特異性GNC抗體SI-39E4、SI-39E2及SI-39E3展示類似活性(第19圖)。以靜止(無功能) FITC結構域替換PD-L1及4-1BB結構域之四特異性GNC抗體SI-39E1及SI-39E5展示溶解活性之減小。此觀察結果確認4-1BB及PD-L1結構域之功能貢獻。
實例
17.
回應於用靶向
EGFRvIII
之四特異性
GNC
抗體進行的治療、
AgBD
位置對
EC50
之值的效應的藉由
PBMC
的顆粒酶
B
產生。
評定表5中列出的一組四特異性及靶向EGFRvIII之GNC抗體藉由PBMC誘導顆粒酶B分泌之能力。用測試分子培養PBMC 96 h。收集上清液且使用ELISA (R&D Systems)分析顆粒酶B之存在,且將顆粒酶B之位準繪圖以判定每一四特異性GNC抗體之EC50。表6列出每一四特異性GNC抗體中AgBD之結構位置。如表6所示,此項研究中活性最大分子全部含有相鄰CD3及PD-L1結構域及4-1BB x TAA (此項研究中為EGFRvIII)。顆粒酶B分泌之此高位準可不為合乎需要的,因為活體內細胞毒性可變得過高。在此情形中,展示最適度但小少20倍之活性的下一組分子SI-39E29及SI-39E18含有相鄰CD3及TAA (此項研究中為EGFRvIII)。
實例
18.
回應於用靶向
ROR1
之四特異性
GNC
抗體進行的治療的針對
Kasumi-2
靶細胞系之重導引
panT
細胞活性。
評定表7中列出的一組四特異性GNC抗體及表4中之SI-35E20溶解靶細胞Kasumi-2之能力。Kasumi-2細胞系穩定地表現經由慢病毒轉導(Clontech)遞送的綠色螢光蛋白質(Green Fluorescent Protein;GFP)。將Kasumi-2腫瘤細胞用panT細胞共培養。經由計數在用panT細胞進行96 h共培養之後留在培養物中的活標靶之數量,用流式細胞術(BD LSRFortessa)評定靶細胞溶解(第20圖)。如第4圖-第9圖所示表徵SI-35E20。此所得展示SI-35E20介導的重導引panT細胞活性針對Kasumi-2靶細胞系之功效為可比較的。
實例
19.
回應於用靶向
CD19
之四特異性
GNC
抗體進行的治療的針對
Kasumi-2
靶細胞系之重導引
PBMC T
細胞活性。
評定表8中列出的一組四特異性GNC抗體的溶解靶細胞Kasumi-2之能力。將Kasumi-2-GFP腫瘤細胞用PBMC共培養。經由計數在用PBMC進行8天共培養之後留在培養物中的活標靶之數量,用流式細胞術(BD LSRFortessa)評定靶細胞溶解(第21圖)。在此項研究中,SI-38E17為當中更有效的分子。
實例
20.
回應於用靶向
CD19
之四特異性
GNC
抗體進行的治療之
CD8 T
細胞增殖。
評定表8中列出的一組四特異性GNC抗體的在靶細胞Kasumi-2存在下刺激CD8 T細胞增殖之能力。用CellTrace紫色染料(Thermo Fisher Scientific)標記PBMC。將Kasumi-2-GFP腫瘤細胞用PBMC共培養。經由在8天共培養之後稀釋CellTrace紫色染料,用流式細胞術(BD LSRFortessa)評定CD8 T細胞增殖。兩種四特異性GNC抗體SI-38E17及SI-38E41為在靶細胞存在下刺激CD8 T細胞增殖方面最有效的(第22圖)。該兩種分子由用於CD3及腫瘤抗原之相鄰結合結構域構成。
實例
21.
回應於用靶向
CD19
之四特異性抗體進行的治療的藉由
PBMC
之
IFNγ
產生。
評定表8中列出的一組四特異性GNC抗體的在靶細胞Kasumi-2存在下藉由PBMC誘導IFNγ分泌之能力。用測試分子培養靶細胞及PBMC 8天。收集上清液且使用ELISA (R&D Systems)分析IFNγ之存在。含有相鄰CD3及PD-L1結構域之分子為繼抗體SI-38E5之後在藉由PBMC誘導IFNγ產生方面為最有效的。SI-38E17在此項研究中展示中等活動(第23圖)。
術語「抗體」係廣義使用,且特定而言,涵蓋單一單株抗體(包括促效劑及拮抗劑抗體)、具有多表位特異性之抗體組合物,以及抗體片段(例如,Fab、F(ab')2、及Fv),只要其展現所要生物活性即可。在一些實施例中,抗體可為單株、多株、嵌合、單鏈、雙特異性或雙有效性、模擬、人類及人源化抗體以及其活性片段。結合至已知抗原的分子之活性片段之實例包括Fab、F(ab')2、scFv及Fv片段,包括Fab免疫球蛋白表現文庫之產物及上文提及的任何抗體及片段之表位結合片段。在一些實施例中,抗體可包括免疫球蛋白分子及免疫球蛋白分子之免疫活性部分,該等免疫球蛋白分子亦即為含有免疫特異性結合抗原之結合位點的分子。免疫球蛋白可具有免疫球蛋白分子之任何類型(IgG、IgM、IgD、IgE、IgA及IgY)或類別(IgG1、IgG2、IgG3、IgG4、IgA1及IgA2)或子類。在一個實施例中,抗體可為完整抗體及來源於完整抗體之任何抗原結合片段。典型抗體係指異源四聚蛋白質,其典型地包含兩個重(H)鏈及兩個輕(L)鏈。每一重鏈由重鏈可變結構域(縮寫為VH)及重鏈恆定結構域構成。每一輕鏈由輕鏈可變結構域(縮寫為VL)及輕鏈恆定結構域構成。VH及VL區域可進一步再分成高度變異互補判定區(complementarity determining region;CDR),及稱為框架區(framework region;FR)之更保守區。每一可變結構域(VH或VL)典型地由三個CDR及四個FR構成,該等區係自胺基末端至羧基末端按以下順序佈置:FR1、CDR1、FR2、CDR2、FR3、CDR3、FR4。在輕鏈及重鏈之可變區內,存在與抗原相互作用的結合區。
如本文所使用的術語「單株抗體」係指自實質上同源抗體之群體獲得的抗體,該等實質上同源抗體亦即為包含除可以微小量存在的可能天然存在的突變外皆為相同的群體之個別抗體。單株抗體為高度特異性,其係針對單一抗原位點。此外,與典型地包括針對不同決定位(表位)不同抗體之習知(多株)抗體製劑相對比,每一單株抗體係針對抗原上之單一決定位。除其特異性之外,單株抗體為有利的,原因在於其係藉由融合瘤培養合成,不受其他免疫球蛋白污染。修飾詞「單株」指示如自抗體之實質上同源群體獲得的抗體之特性,且不欲解釋為需要藉由任何特定方法來產生抗體。例如,根據本揭示內容使用的單株抗體可藉由首先由Kohler & Milstein, Nature, 256:495 (1975)描述的融合瘤方法來製造,或可藉由重組DNA方法(參見,例如,美國專利第4,816,567號)來製造。
單株抗體可包括「嵌合」抗體(免疫球蛋白),其中重鏈及/或輕鏈之一部分與來源於特定物種或屬於特定抗體類別或子類之抗體中的對應序列一致或同源,而鏈之剩餘部分係與來源於另一物種或屬於另一抗體類別或子類的抗體以及此種抗體之片段中之對應序列一致或同源,只要其展現所要生物活性即可(美國專利第4,816,567號;及Morrison等人 Proc. Natl. Acad. Sci. USA, 81:6851-6855 [1984])。
單株抗體可使用包括小鼠融合瘤或噬菌體呈現(參見Siegel. Transfus. Clin. Biol. 9:15-22 (2002)之綜述)的各種方法產生,或自直接來自初級B細胞之抗體之分子選殖(參見Tiller. New Biotechnol. 28:453-7 (2011))產生。在本揭示內容中,抗體係藉由用人類PD-L1蛋白質及在細胞表面上短暫表現人類PD-L1之細胞免疫兔子來產生。已知兔子產生高親和力、多樣性及特異性之抗體(Weber等人, Exp. Mol. Med. 49:e305)。來自免疫動物之B細胞係活體外培養及篩選用於產生抗PD-L1抗體。抗體可變基因係使用重組DNA技術分離且所得抗體係重組表現且針對所要特徵來進一步篩選,該等所要特徵諸如抑制PD-L1與PD-1之結合的能力、結合至非人類靈長類動物PD-L1之能力及增強人類T細胞活化之能力。抗體發現之此一般方法係類似於Seeber等人, PLOS One. 9:e86184 (2014)中描述的彼方法。
術語「抗原或表位結合部分或片段」係指抗體的能夠結合至抗原(在此情況下為PD-L1)之片段。該些片段可能夠具有完整抗體之抗原結合功能及另外的功能。結合片段之實例包括但不限於單鏈Fv片段(scFv),其由藉由合成連接子或Fab片段連接在單一多肽鏈中的抗體之單臂之VL及VH結構域組成,該合成連接子或Fab片段為由VL、恆定輕鏈(CL)、VH及恆定重鏈1 (CH1)結構域組成之一價片段。抗體片段可甚至為較小子片段且可由小達單一CDR結構域、尤其為來自VL及/或VH結構域之CDR3區的結構域組成(例如,參見Beiboer等人, J. Mol. Biol. 296:833-49 (2000))。使用熟習此項技術者已知的習知方法產生抗體片段。抗體片段可使用在完整抗體情況下使用的相同技術來針對實用性進行篩選。
「抗原或表位結合片段」可藉由多種此項技術中已知的技術自本揭示內容之抗體得到。例如,純化的單株抗體可利用諸如胃蛋白酶之酶來分解,且經受HPLC凝膠過濾。可隨後收集含有Fab片段之適當餾分且藉由膜過濾及類似物來濃縮。對用於分離抗體之活性片段的一般技術之進一步描述,參見例如,Khaw, B. A.等人, J. Nucl. Med. 23:1011-1019 (1982);Rousseaux等人, Methods Enzymology, 121:663-69, Academic Press, 1986。
抗體之木瓜酶消化產生兩個一致的抗原結合片段,稱為「Fab」片段,每一片段具有單一抗原結合位點,及殘留「Fc」片段,該片段之名稱反映其容易結晶之能力。胃蛋白酶處理產生F(ab')2片段,其具有兩個抗原組合位點且仍能夠交聯抗原。
Fab片段可含有輕鏈之恆定結構域及重鏈之第一恆定結構域(CH1)。因在重鏈CH1結構域之羧基終點處添加包括來自抗體鉸鏈區之一或多個半胱胺酸的少數殘基,Fab'片段不同於Fab片段。Fab'-SH為在本文對Fab'之命名,其中恆定結構域之半胱胺酸殘基帶有游離的硫醇基。F(ab')2抗體片段原始地係產生問問成對的Fab'片段,其之間具有鉸鏈半胱胺酸。亦已知抗體片段之其他、化學偶合。
「Fv」為最小抗體片段,其含有完全抗原識別及結合位點。此區由呈緊密、非共價締合的一個重鏈可變結構域及一個輕鏈可變結構域之二聚物組成。在此配置中,每一可變結構域之三個CDR相互作用以在VH-VL二聚物之表面上定義抗原結合位點。共同地,六個CDR將抗原結合特異性賦予抗體。然而,甚至單一可變結構域(或Fv之一半,其包含僅三個對抗原為特異性的CDR)具有辨別並結合抗原之能力,但以比完整結合位點低的親和力達成。
來自任何脊椎動物物種的抗體(免疫球蛋白)之「輕鏈」可基於其恆定結構域之胺基酸序列而指定為稱為κ及λ(A)的兩個明顯相異類型之一。
取決於其重鏈之恆定結構域之胺基酸序列,免疫球蛋白可指定至不同類別。存在五個主要類別之免疫球蛋白:IgA、IgD、IgE、IgG及IgM,且該些免疫球蛋白中若干者可進一步分成子類(同型),例如,IgG-1、IgG-2、IgG-3、及IgG-4;IgA-1及IgA-2。對應於不同類別之免疫球蛋白的重鏈恆定結構域分別稱為α、δ、ε、γ、及μ。不同類別的免疫球蛋白之子單元結構及三維配置。
「人源化抗體」係指一類工程化抗體,其具有其來源於非人類供體免疫球蛋白之CDR,分子之剩餘免疫球蛋白源性部分係來源於一種(或多種)人免疫球蛋白。另外,框架支撐殘基可經改變以保存結合親和力。獲得「人源化抗體」之方法係熟習此項技術者熟知的。(參見,例如,Queen等人, Proc. Natl Acad Sci USA, 86:10029-10032 (1989),Hodgson等人, Bio/Technology, 9:421 (1991))。在一個實施例中,「人源化抗體」可藉由遺傳工程化方法來獲得,該等遺傳工程化方法賦能親和力成熟類人類多株抗體於諸如例如兔子之大型動物中之產生(參見,例如美國專利第7,129,084號)。
如本文所使用的術語「多肽」、「肽」、及「蛋白質」係可互換的且經定義來意指由藉由肽鍵連接的胺基酸構成的生物分子。
如本文所使用術語「一(a/an)」及「該」係定義來意指「一或多個」且包括複數,除非情景係不適當的。
「分離的」意指生物分子,其不含至少一些與其天然存在的組分。當用於描述本文揭示的各種多肽時,「分離」意指已經鑑別且自表現其之細胞或細胞培養物分離及/或回收的多肽。通常,分離多肽將藉由至少一個純化步驟來製備。「分離抗體」係指實質上不含具有不同抗原特異性之其他抗體的抗體。
「重組」意指抗體係使用重組核酸技術在外來寄主細胞中產生。
術語「抗原」係指可在生物體、特定而言動物、更特定而言包括人類之哺乳動物中誘導免疫反應的實體或其片段。術語包括負責抗原性或抗原決定位之免疫原及其區域。
亦如本文所使用,術語「免疫原性」係指引出或增強抗體、T細胞或針對免疫原性劑的其他反應性免疫細胞之產生且貢獻於人類或動物中之免疫反應的物質。當個體產生充分的抗體、T細胞及抵抗所投與的本揭示內容之免疫原性組合物的其他反應性免疫細胞以緩和或減輕將要治療的病症時,免疫反應就發生。
「特異性結合」或「特異地結合至」或「對特定抗原或表位為特異性」意指適度地不同於非特異性相互作用之結合。特異性結合可例如藉由相較於對照分子之結合測定分子之結合來量測,該對照分子通常為不具有結合活性的具有類似結構之分子。例如,特異性結合可藉由與類似於標靶的對照分子競爭來測定。
用於特定抗原或表位之特異性結合可例如藉由具有至少約10-4 M、至少約10-5 M、至少約10-6 M、至少約10-7 M、至少約10-8 M、至少約10-9、替代地至少約10-10 M、至少約10-11 M、至少約10-12 M、或更大的對抗原或表位之KD的抗體來展現,其中KD係指特定抗體-抗原相互作用之解離速率。在一些實施例中,特異地結合抗原的抗體將具有比對照分子相對於抗原或表位大20倍、50倍、100倍、500倍、1000倍、5,000倍、10,000倍或更多倍的KD。
此外,對特定抗原或表位之特異性結合可例如藉由具有對抗原或表位比相對於對照對表位大至少20倍、50倍、100倍、500倍、1000倍、5,000倍、10,000倍或更多倍的KA或Ka之抗體來展現,其中KA或Ka係指特定抗體-抗原相互作用之締合速率。
兩個序列之間的「同源性」係藉由序列一致性來測定。若彼此比較的兩個序列在長度上有所不同,則序列一致性較佳地係關於較短序列中與較長序列之核苷酸殘基一致的核苷酸殘基之百分比。序列一致性可按慣例使用電腦程式來測定。在給定序列與本揭示內容之上述序列之間的比較中出現的偏差可例如藉由添加、缺失、替代、插入或重組來引起。
儘管本揭示內容已參考特定實施例或實例描述,但可理解的是,實施例為說明性的且本揭示內容之範疇不因此受限。本揭示內容之替代實施例可對熟習本揭示內容所屬技術領域之一般技藝人士變得明顯。此種替代實施例係視為涵蓋在本揭示內容之範疇內。因此,本揭示內容之範疇係藉由隨附申請專利範圍界定且由前述描述來支援。本揭示內容中引用或參考的所有參考文獻之全部內容係在此以全文引用方式併入。表
四特異性
GNC
抗體之序列表
胺基酸序列中加底線之CDR
>SEQ ID 01 抗CD3 284A10 VHv1 nt
GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCA
>SEQ ID 02 抗CD3 284A10 VHv1 aa
EVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNN
IWGQGTLVTVSS
>SEQ ID 03 抗CD3 284A10 VLv1 nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 04 抗CD3 284A10 VLv1 aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIK
>SEQ ID 05 抗CD3 480C8 VHv1 nt
GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGAATCGACCTCAGTAGCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTAATAGTAAGAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCA
>SEQ ID 06 抗CD3 480C8 VHv1 aa
EVQLVESGGGLVQPGGSLRLSCAASGIDLSSNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAINSKNI
WGQGTLVTVSS
>SEQ ID 07 抗CD3 480C8 VLv1 nt
GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 08 抗CD3 480C8 VLv1 aa
DIQMTQSPSTLSASVGDRVTITCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNA
FGGGTKVEIK
>SEQ ID 09 抗PD-L1 PL230C6 VHv3 nt
CAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCCAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGT
>SEQ ID 10 抗PD-L1 PL230C6 VHv3 aa
QSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMI
WVRQAPGKGLEWVGIITYSGSRYYANWAKG
RFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHL
WGQGTLVTVSS
>SEQ ID 11 抗PD-L1 PL230C6 VLv2 nt
GCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 12 抗PD-L1 PL230C6 VLv2 aa
AYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWY
QQKPGKAPKLLIHSASSLAS
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNA
FGGGTKVEIK
>SEQ ID 13 抗4-1BB 420H5 VHv3 nt
CAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCAACTACTGGATATGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTTATGTTGGTAGTAGTGGTGACACTTACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGATAGTAGTAGTTATTATATGTTTAACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC
>SEQ ID 14 抗4-1BB 420H5 VHv3 aa
QSLVESGGGLVQPGGSLRLSCAASGFSFSSNYWIC
WVRQAPGKGLEWIACIYVGSSGDTYYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSSSYYMFNL
WGQGTLVTVSS
>SEQ ID 15 抗4-1BB 420H5 VLv3 nt
GCCCTTGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTGAGGACATTGATACCTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTTATGCATCCGATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCGGTTACTATACTAGTAGTGCTGATACGAGGGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 16 抗4-1BB 420H5 VLv3 aa
ALVMTQSPSTLSASVGDRVTINCQASEDIDTYLA
WYQQKPGKAPKLLIFYASDLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGGYYTSSADTRGA
FGGGTKVEIK
>SEQ ID 17 抗4-1BB 466F6 VHv2 nt
CGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC
>SEQ ID 18 抗4-1BB 466F6 VHv2 aa
RSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSS
>SEQ ID 19 抗4-1BB 466F6 VLv5 nt
GACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 20 抗4-1BB 466F6 VLv5 aa
DVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIK
>SEQ ID 21 抗4-1BB 460C3 VHv1 nt
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGAATCGACTTCAGTAGGAGATACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATATATACTGGTAGCCGCGATACTCCTCACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGAAGGTAGCCTGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC
>SEQ ID 22 抗4-1BB 460C3 VHv1 aa
EVQLLESGGGLVQPGGSLRLSCAASGIDFSRRYYMC
WVRQAPGKGLEWIACIYTGSRDTPHYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREGSL
WGQGTLVTVSS
>SEQ ID 23 抗4-1BB 460C3 VLv1 nt
GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAGTAACTGGTTCTCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCGCAGGCGGTTACAATACTGTTATTGATACTTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 24 抗4-1BB 460C3 VLv1 aa
DIQMTQSPSTLSASVGDRVTITCQSSQSVYSNWFS
WYQQKPGKAPKLLIYSASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCAGGYNTVIDTFA
FGGGTKVEIK
>SEQ ID 25 抗ROR1 324C6 VHv2 nt
CAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCTCCCTCAGTAGGTACTACATGACCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAACCATTTATACTAGTGGTAGTACATGGTACGCGAGCTGGACAAAAGGCAGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGATCCTATTATGGCGGTGATAAGACTGGTTTAGGCATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCA
>SEQ ID 26 抗ROR1 324C6 VHv2 nt
QSLVESGGGLVQPGGSLRLSCTASGFSLSRYYMT
WVRQAPGKGLEWIGTIYTSGSTWYASWTKG
RFTISKDNTKNTVDLQMNSLRAEDTAVYYCARSYYGGDKTGLGI
WGQGTLVTVSS
>SEQ ID 27 抗ROR1 324C6 VLv1 nt
GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTGATAGTTGGTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATCAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAATCTGCTTATGGTGTTAGTGGTACTAGTAGTTATTTATATACTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 28 抗ROR1 324C6 VLv1 aa
DIQMTQSPSTLSASVGDRVTITCQASQSIDSWLS
WYQQKPGKAPKLLIYQASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQSAYGVSGTSSYLYT
FGGGTKVEIK
>SEQ ID 29 抗ROR1 323H7 VHv4 nt
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCA
>SEQ ID 30 抗ROR1 323H7 VHv4 aa
EVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMT
WVRQAPGKGLEWIGHIYVNNDDTDYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDI
WGQGTLVTVSS
>SEQ ID 31 抗ROR1 323H7 VLv1 nt
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 32 抗ROR1 323H7 VLv1 aa
DIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLA
WYQQKPGKVPKLLIYYASTLAS
GVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFA
FGGGTKVEIK
>SEQ ID 33 抗ROR1 338H4 VHv3 nt
GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCTCCCTCAGTAGCTATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAGGGGGCTGGAGTGGATCGGAATCATTTATGCTAGTGGTAGCACATACTACGCGAGCTCGGCGAAAGGCAGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAATTTATGACGGCATGGACCTCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCA
>SEQ ID 34 抗ROR1 338H4 VHv3 aa
EVQLVESGGGLVQPGGSLRLSCTASGFSLSSYAMS
WVRQAPGRGLEWIGIIYASGSTYYASSAKG
RFTISKDNTKNTVDLQMNSLRAEDTAVYYCARIYDGMDL
WGQGTLVTVSS
>SEQ ID 35 抗ROR1 338H4 VLv4 nt
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTCAGAACATTTACAGCTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCGCCTGATCTATCTGGCATCTACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTACACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTCAAAGCAATTATAACGGTAATTATGGTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 36 抗ROR1 338H4 VLv4 aa
DIQMTQSPSSLSASVGDRVTINCQASQNIYSYLS
WYQQKPGKVPKRLIYLASTLAS
GVPSRFSGSGSGTDYTLTISSLQPEDVATYYCQSNYNGNYG
FGGGTKVEIK
>SEQ ID 37 抗ROR1 330F11 VHv1 nt
GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCCTCAATAACTACTGGATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAACCATTAGTAGTGGTGCGTATACATGGTTCGCCACCTGGGCGACAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGATATTCTTCTACTACTGATTGGACCTACTTTAACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCA
>SEQ ID 38 抗ROR1 330F11 VHv1 aa
EVQLVESGGGLVQPGGSLRLSCAASGFSLNNYWMS
WVRQAPGKGLEWIGTISSGAYTWFATWATG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARYSSTTDWTYFNI
WGQGTLVTVSS
>SEQ ID 39 抗ROR1 330F11 VLv1 nt
GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAATAACTACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAGGGCATCCACTCTGGAATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAAGCTATAATGGTGTTGGTAGGACTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 40 抗ROR1 330F11 VLv1 aa
DIQMTQSPSTLSASVGDRVTITCQASQSINNYLA
WYQQKPGKAPKLLIYRASTLES
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQSYNGVGRTA
FGGGTKVEIK
>SEQ ID 41 抗FITC 4-4-20 VH nt
GAGGTGAAGCTGGATGAGACTGGAGGAGGCTTGGTGCAACCTGGGAGGCCCATGAAACTCTCCTGTGTTGCCTCTGGATTCACTTTTAGTGACTACTGGATGAACTGGGTCCGCCAGTCTCCAGAGAAAGGACTGGAGTGGGTAGCACAAATTAGAAACAAACCTTATAATTATGAAACATATTATTCAGATTCTGTGAAAGGCAGATTCACCATCTCAAGAGATGATTCCAAAAGTAGTGTCTACCTGCAAATGAACAACTTAAGAGTTGAAGACATGGGTATCTATTACTGTACGGGTTCTTACTATGGTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA
>SEQ ID 42 抗FITC 4-4-20 VH aa
EVKLDETGGGLVQPGRPMKLSCVASGFTFSDYWMN
WVRQSPEKGLEWVAQIRNKPYNYETYYSDSVKG
RFTISRDDSKSSVYLQMNNLRVEDMGIYYCTGSYYGMDY
WGQGTSVTVSS
>SEQ ID 43 抗FITC 4-4-20 VL nt
GATGTCGTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATCTAGTCAGAGCCTTGTACACAGTAATGGAAACACCTATTTACGTTGGTACCTGCAGAAGCCAGGCCAGTCTCCAAAGGTCCTGATCTACAAAGTTTCCAACCGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAAGATCAGCAGAGTGGAGGCTGAGGATCTGGGAGTTTATTTCTGCTCTCAAAGTACACATGTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA
>SEQ ID 44 抗FITC 4-4-20 VL aa
DVVMTQTPLSLPVSLGDQASISCRSSQSLVHSNGNTYLR
WYLQKPGQSPKVLIYKVSNRFS
GVPDRFSGSGSGTDFTLKISRVEAEDLGVYFCSQSTHVPWT
FGGGTKLEIK
>SEQ ID 45 人類IgG1 null (具有ADCC/CDC無效突變之G1m-fa) nt
GCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGT
>SEQ ID 46 人類IgG1 null (具有ADCC/CDC無效突變之G1m-fa) aa
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG
>SEQ ID 47 人類Ig κ nt
CGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 48 人類Ig κ aa
RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 49 SI-35E18 (460C3-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)重鏈nt
GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAGTAACTGGTTCTCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATTCTGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCGCAGGCGGTTACAATACTGTTATTGATACTTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGAATCGACTTCAGTAGGAGATACTACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATATATACTGGTAGCCGCGATACTCCTCACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGAAGGTAGCCTGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCCAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 50 SI-35E18 (460C3-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)重鏈aa
DIQMTQSPSTLSASVGDRVTITCQSSQSVYSNWFSWYQQKPGKAPKLLIYSASTLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCAGGYNTVIDTFAFGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGIDFSRRYYMCWVRQAPGKGLEWIACIYTGSRDTPHYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAREGSLWGQGTLVTVSSGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMIWVRQAPGKGLEWVGIITYSGSRYYANWAKGRFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHLWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMTWVRQAPGKGLEWIGHIYVNNDDTDYASSAKGRFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLAWYQQKPGKVPKLLIYYASTLASGVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFAFGGGTKVEIKGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMSWVRQAPGKGLEWIGVITGRDITYYASWAKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNIWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSTLSASVGDRVTINCQASESISSWLAWYQQKPGKAPKLLIYEASKLASGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNSFGGGTKVEIK
>SEQ ID 51 SI-35E18 (460C3-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)輕鏈nt
GCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 52 SI-35E18 (460C3-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)輕鏈aa
AYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLAWYQQKPGKAPKLLIHSASSLASGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNAFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 53 抗CD3 284A10 VHv1b nt
GAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACA
>SEQ ID 54 抗CD3 284A10 VHv1b aa
EVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVST
>SEQ ID 55 抗4-1BB 466F6b VHv2 nt
CGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTGTCGACA
>SEQ ID 56 抗4-1BB 466F6b VHv2 aa
RSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVST
>SEQ ID 57 抗PD-L1 PL230C6 VHv3b nt
CAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACCGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTGTCGACA
>SEQ ID 58 抗PD-L1 PL230C6 VHv3b aa
QSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMI
WVRQAPGKGLEWVGIITYSGSRYYANWAKG
RFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHL
WGQGTLVTVST
>SEQ ID 59 抗huPD-L1 PL221G5 VHv1 nt
GAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGC
>SEQ ID 60 抗huPD-L1 PL221G5 VHv1 aa
EVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMC
WVRQAPGKGLEWIACIAAGSAGITYDANWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDL
WGQGTLVTVSS
>SEQ ID 61 抗huPD-L1 PL221G5 VLv1 nt
GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 62 抗huPD-L1 PL221G5 VLv1 aa
DIQMTQSPSTLSASVGDRVTITCQASQSISSHLN
WYQQKPGKAPKLLIYKASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNV
FGGGTKVEIK
>SEQ ID 63 抗huCD19 21D4 VH nt
GAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAGAAACCAGGAGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGCTTTAGCAGTTCATGGATCGGCTGGGTGCGCCAGGCACCTGGGAAAGGCCTGGAATGGATGGGGATCATCTATCCTGATGACTCTGATACCAGATACAGTCCATCCTTCCAAGGCCAGGTCACCATCTCAGCCGACAAGTCCATCAGGACTGCCTACCTGCAGTGGAGTAGCCTGAAGGCCTCGGACACCGCTATGTATTACTGTGCGAGACATGTTACTATGATTTGGGGAGTTATTATTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA
>SEQ ID 64 抗huCD19 21D4 VH aa
EVQLVQSGAEVKKPGESLKISCKGSGYSFSSSWIG
WVRQAPGKGLEWMGIIYPDDSDTRYSPSFQG
QVTISADKSIRTAYLQWSSLKASDTAMYYCARHVTMIWGVIIDF
WGQGTLVTVSS
>SEQ ID 65 抗huCD19 21D4 VL nt
GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAA
>SEQ ID 66 抗huCD19 21D4 VL aa
AIQLTQSPSSLSASVGDRVTITCRASQGISSALA
WYQQKPGKAPKLLIYDASSLES
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPFT
FGPGTKVDIK
>SEQ ID 67 抗huEGFRvIII 806 VH nt
GATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA
>SEQ ID 68 抗huEGFRvIII 806 VH aa
DVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWN
WIRQFPGNKLEWMGYISYSGNTRYNPSLKS
RISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPY
WGQGTLVTVSA
>SEQ ID 69 抗huEGFRvIII 806 VL nt
GACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA
>SEQ ID 70 抗huEGFRvIII 806 VL aa
DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIG
WLQQRPGKSFKGLIYHGTNLDD
EVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWT
FGGGTKLEIK
>SEQ ID 71 GGGGSGGGGSG連接子nt
GGCGGTGGAGGGTCCGGCGGTGGTGGCTCCGGA
>SEQ ID 72 GGGGSGGGGSG連接子aa
GGGGSGGGGSG
>SEQ ID 73 GGGGSGGGGS連接子01 nt
GGCGGTGGAGGGTCCGGCGGTGGTGGATCA
>SEQ ID 74 GGGGSGGGGS連接子01 aa
GGGGSGGGGS
>SEQ ID 75 GGGGSGGGGS連接子02 nt
GGCGGTGGAGGGTCCGGCGGTGGTGGATCC
>SEQ ID 76 GGGGSGGGGS連接子02 aa
GGGGSGGGGS
>SEQ ID 77 GGGGSGGGGSGGGGSGGGGS連接子nt
GGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCA
>SEQ ID 78 GGGGSGGGGSGGGGSGGGGS連接子aa
GGGGSGGGGSGGGGSGGGGS
>SEQ ID 79 SI-39E18 (284A10-L1H1-scFv x 806-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)重鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCAACTACTGGATATGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGTATTTATGTTGGTAGTAGTGGTGACACTTACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGATAGTAGTAGTTATTATATGTTTAACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGCCCTTGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTGAGGACATTGATACCTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTTACGCATCCGATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCGGTTACTATACTAGTAGTGCTGATACGAGGGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 80 SI-39E18 (284A10-L1H1-scFv x 806-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)重鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWN
WIRQFPGNKLEWMGYISYSGNTRYNPSLKS
RISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPY
WGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMC
WVRQAPGKGLEWIACIAAGSAGITYDANWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLN
WYQQKPGKAPKLLIYKASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNV
FGGGTKVEIKGGGGSGGGGSQSLVESGGGLVQPGGSLRLSCAASGFSFSSNYWIC
WVRQAPGKGLEWIACIYVGSSGDTYYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSSSYYMFNL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSALVMTQSPSTLSASVGDRVTINCQASEDIDTYLA
WYQQKPGKAPKLLIFYASDLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGGYYTSSADTRGA
FGGGTKVEIK
>SEQ ID 81 SI-39E18 (284A10-L1H1-scFv x 806-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)輕鏈nt
GACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 82 SI-39E18 (284A10-L1H1-scFv x 806-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)輕鏈aa
DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIG
WLQQRPGKSFKGLIYHGTNLDD
EVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWT
FGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 83 SI-39E29 (806-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)重鏈nt
GACATCCTGATGACCCAATCTCCATCCTCCATGTCTGTATCTCTGGGAGACACAGTCAGCATCACTTGCCATTCAAGTCAGGACATTAACAGTAATATAGGGTGGTTGCAGCAGAGACCAGGGAAATCATTTAAGGGCCTGATCTATCATGGAACCAACTTGGACGATGAAGTTCCATCAAGGTTCAGTGGCAGTGGATCTGGAGCCGATTATTCTCTCACCATCAGCAGCCTGGAATCTGAAGATTTTGCAGACTATTACTGTGTACAGTATGCTCAGTTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGATGTGCAGCTTCAGGAGTCGGGACCTAGCCTGGTGAAACCTTCTCAGTCTCTGTCCCTCACCTGCACTGTCACTGGCTACTCAATCACCAGTGATTTTGCCTGGAACTGGATTCGGCAGTTTCCAGGAAACAAGCTGGAGTGGATGGGCTACATAAGTTATAGTGGTAACACTAGGTACAACCCATCTCTCAAAAGTCGAATCTCTATCACTCGCGACACATCCAAGAACCAATTCTTCCTGCAGTTGAACTCTGTGACTATTGAGGACACAGCCACATATTACTGTGTAACGGCGGGACGCGGGTTTCCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCAACTACTGGATATGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGTATTTATGTTGGTAGTAGTGGTGACACTTACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGAGATAGTAGTAGTTATTATATGTTTAACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGCCCTTGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAGGCCAGTGAGGACATTGATACCTATTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTTTTACGCATCCGATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCGGTTACTATACTAGTAGTGCTGATACGAGGGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 84 SI-39E29 (806-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)重鏈aa
DILMTQSPSSMSVSLGDTVSITCHSSQDINSNIG
WLQQRPGKSFKGLIYHGTNLDD
EVPSRFSGSGSGADYSLTISSLESEDFADYYCVQYAQFPWT
FGGGTKLEIKGGGGSGGGGSGGGGSGGGGSDVQLQESGPSLVKPSQSLSLTCTVTGYSITSDFAWN
WIRQFPGNKLEWMGYISYSGNTRYNPSLKS
RISITRDTSKNQFFLQLNSVTIEDTATYYCVTAGRGFPY
WGQGTLVTVSAGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMC
WVRQAPGKGLEWIACIAAGSAGITYDANWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLN
WYQQKPGKAPKLLIYKASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNV
FGGGTKVEIKGGGGSGGGGSQSLVESGGGLVQPGGSLRLSCAASGFSFSSNYWIC
WVRQAPGKGLEWIACIYVGSSGDTYYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDSSSYYMFNL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSALVMTQSPSTLSASVGDRVTINCQASEDIDTYLA
WYQQKPGKAPKLLIFYASDLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGGYYTSSADTRGA
FGGGTKVEIK
>SEQ ID 85 SI-39E29 (806-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)輕鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 86 SI-39E29 (806-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 420H5-H3L3-scFv)輕鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 87 SI-35E20 (466F6-L5H2-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)重鏈nt
GACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCCAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTAGTGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 88 SI-35E20 (466F6-L5H2-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)重鏈aa
DVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIKGGGGSGGGGSGGGGSGGGGSRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSSGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMI
WVRQAPGKGLEWVGIITYSGSRYYANWAKG
RFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHL
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMT
WVRQAPGKGLEWIGHIYVNNDDTDYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDI
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLA
WYQQKPGKVPKLLIYYASTLAS
GVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFA
FGGGTKVEIKGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIK
>SEQ ID 89 SI-35E20 (466F6-L5H2-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)輕鏈nt
GCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 90 SI-35E20 (466F6-L5H2-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 284A10-H1L1-scFv)輕鏈aa
AYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLA
WYQQKPGKAPKLLIHSASSLAS
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNA
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 91 SI-35E58 (284A10-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 466F6-H2L5-scFv)重鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACCGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGGTCCGGAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCTTCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGGTCCGGACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 92 SI-35E58 (284A10-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 466F6-H2L5-scFv)重鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSTGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMI
WVRQAPGKGLEWVGIITYSGSRYYANWAKG
RFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHL
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSGEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMT
WVRQAPGKGLEWIGHIYVNNDDTDYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDI
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLA
WYQQKPGKVPKLLIYYASTLAS
GVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFA
FGGGTKVEIKGGGGSGGGGSGRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIK
>SEQ ID 93 SI-35E58 (284A10-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 466F6-H2L5-scFv)輕鏈nt
GCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 94 SI-35E58 (284A10-L1H1-scFv x PL230C6-Fab x 323H7-H4L1-scFv x 466F6-H2L5-scFv)輕鏈aa
AYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLA
WYQQKPGKAPKLLIHSASSLAS
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNA
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 95 SI-35E88 (284A10-L1H1-scFv x 323H7-Fab x PL230C6-H3L2-scFv x 466F6-H2L5-scFv)重鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCAGTCGGTGGAGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACCGCCTCTGGAATCGACCTTAATACCTACGACATGATCTGGGTCCGCCAGGCTCCAGGCAAGGGGCTAGAGTGGGTTGGAATCATTACTTATAGTGGTAGTAGATACTACGCGAACTGGGCGAAAGGCCGATTCACCATCTCCAAAGACAATACCAAGAACACGGTGTATCTGCAAATGAACAGCCTGAGAGCTGAGGACACGGCTGTGTATTACTGTGCGAGAGATTATATGAGTGGTTCCCACTTGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCCGGTGGAGGCGGTTCAGGCGGAGGTGGAAGTGGTGGTGGCGGCTCTGGAGGCGGCGGATCTGCCTATGATATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCAAGTGTCAGGCCAGTGAGGACATTTATAGCTTCTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCCATTCTGCATCCTCTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTACTATTGTCAACAGGGTTATGGTAAAAATAATGTTGATAATGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGGTCCGGACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 96 SI-35E88 (284A10-L1H1-scFv x 323H7-Fab x PL230C6-H3L2-scFv x 466F6-H2L5-scFv)重鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSTGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMT
WVRQAPGKGLEWIGHIYVNNDDTDYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDI
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSQSVEESGGGLVQPGGSLRLSCTASGIDLNTYDMI
WVRQAPGKGLEWVGIITYSGSRYYANWAKG
RFTISKDNTKNTVYLQMNSLRAEDTAVYYCARDYMSGSHL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSAYDMTQSPSSVSASVGDRVTIKCQASEDIYSFLA
WYQQKPGKAPKLLIHSASSLAS
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGYGKNNVDNA
FGGGTKVEIKGGGGSGGGGSGRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIK
>SEQ ID 97 SI-35E88 (284A10-L1H1-scFv x 323H7-Fab x PL230C6-H3L2-scFv x 466F6-H2L5-scFv)輕鏈nt
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 98 SI-35E88 (284A10-L1H1-scFv x 323H7-Fab x PL230C6-H3L2-scFv x 466F6-H2L5-scFv)輕鏈aa
DIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLA
WYQQKPGKVPKLLIYYASTLAS
GVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFA
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 99 SI-35E99 (284A10-L1H1-scFv x 323H7-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)重鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACGGTGGTTCTTCTGCTATTACTAGTAACAACATTTGGGGCCAGGGAACCCTGGTCACCGTGTCGACAGGCGGTGGAGGGTCCGGCGGTGGTGGATCAGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTCGCTACCACATGACTTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGACATATTTATGTTAATAATGATGACACAGACTACGCGAGCTCCGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCACCTATTTCTGTGCGAGATTGGATGTTGGTGGTGGTGGTGCTTATATTGGGGACATCTGGGGCCAGGGAACTCTGGTTACCGTCTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGTGGAGGCGGATCTGGCGGAGGTGGTTCCGGCGGTGGCGGCTCCGGTGGAGGCGGCTCTGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGCTCCGGACGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACTGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCAAGCTCCGCTAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCTTCAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 100 SI-35E99 (284A10-L1H1-scFv x 323H7-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)重鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSTGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFTISRYHMT
WVRQAPGKGLEWIGHIYVNNDDTDYASSAKG
RFTISRDNSKNTLYLQMNSLRAEDTATYFCARLDVGGGGAYIGDI
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMC
WVRQAPGKGLEWIACIAAGSAGITYDANWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLN
WYQQKPGKAPKLLIYKASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNV
FGGGTKVEIKGGGGSGGGGSGRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIK
>SEQ ID 101 SI-35E99 (284A10-L1H1-scFv x 323H7-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)輕鏈nt
GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGTCCAGTCAGAGTGTTTATAACAACAACGACTTAGCCTGGTATCAGCAGAAACCAGGGAAAGTTCCTAAGCTCCTGATCTATTATGCATCCACTCTGGCATCTGGGGTCCCATCTCGGTTCAGTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATGTTGCAACTTATTACTGTGCAGGCGGTTATGATACGGATGGTCTTGATACGTTTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 102 SI-35E99 (284A10-L1H1-scFv x 323H7-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)輕鏈aa
DIQMTQSPSSLSASVGDRVTITCQSSQSVYNNNDLA
WYQQKPGKVPKLLIYYASTLAS
GVPSRFSGSGSGTDFTLTISSLQPEDVATYYCAGGYDTDGLDTFA
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 103 SI-38E17 (284A10-L1H1-scFv x 21D4-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)重鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAGTTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAGAAACCAGGAGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGCTTTAGCAGTTCATGGATCGGCTGGGTGCGCCAGGCACCTGGGAAAGGCCTGGAATGGATGGGGATCATCTATCCTGATGACTCTGATACCAGATACAGTCCATCCTTCCAAGGCCAGGTCACCATCTCAGCCGACAAGTCCATCAGGACTGCCTACCTGCAGTGGAGTAGCCTGAAGGCCTCGGACACCGCTATGTATTACTGTGCGAGACATGTTACTATGATTTGGGGAGTTATTATTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGTGGAGGCGGATCTGGCGGAGGTGGTTCCGGCGGTGGCGGCTCCGGTGGAGGCGGCTCTGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 104 SI-38E17 (284A10-L1H1-scFv x 21D4-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)重鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSSGGGGSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFSSSWIG
WVRQAPGKGLEWMGIIYPDDSDTRYSPSFQG
QVTISADKSIRTAYLQWSSLKASDTAMYYCARHVTMIWGVIIDF
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMC
WVRQAPGKGLEWIACIAAGSAGITYDANWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLN
WYQQKPGKAPKLLIYKASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNV
FGGGTKVEIKGGGGSGGGGSRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIK
>SEQ ID 105 SI-38E17 (284A10-L1H1-scFv x 21D4-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)輕鏈nt
GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 106 SI-38E17 (284A10-L1H1-scFv x 21D4-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)輕鏈aa
AIQLTQSPSSLSASVGDRVTITCRASQGISSALA
WYQQKPGKAPKLLIYDASSLES
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPFT
FGPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
>SEQ ID 107 SI-38E33 (21D4-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)重鏈nt
GCCATCCAGTTGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCGGGCAAGTCAGGGCATTAGCAGTGCTTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCTCCTAAGCTCCTGATCTATGATGCCTCCAGTTTGGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCAGCCTGCAGCCTGAAGATTTTGCAACTTATTACTGTCAACAGTTTAATAGTTACCCATTCACTTTCGGCCCTGGGACCAAAGTGGATATCAAAGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGAGGTGCAGCTGGTGCAGTCTGGAGCAGAGGTGAAGAAACCAGGAGAGTCTCTGAAGATCTCCTGTAAGGGTTCTGGATACAGCTTTAGCAGTTCATGGATCGGCTGGGTGCGCCAGGCACCTGGGAAAGGCCTGGAATGGATGGGGATCATCTATCCTGATGACTCTGATACCAGATACAGTCCATCCTTCCAAGGCCAGGTCACCATCTCAGCCGACAAGTCCATCAGGACTGCCTACCTGCAGTGGAGTAGCCTGAAGGCCTCGGACACCGCTATGTATTACTGTGCGAGACATGTTACTATGATTTGGGGAGTTATTATTGACTTCTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCATCAGTACCAATGCAATGAGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGGAGTCATTACTGGTCGTGATATCACATACTACGCGAGCTGGGCGAAAGGCAGATTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGCGCGACGGTGGATCATCTGCTATTACTAGTAACAACATTTGGGGCCAAGGAACTCTGGTCACCGTTTCTTCAGCTAGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCGGTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGCACCTGAAGCCGCGGGGGCACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCGCGGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTATACCCTGCCCCCATCCCGGGATGAGCTGACCAAGAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGTGGCGGTGGAGGGTCCGGCGGTGGTGGATCCGAGGTGCAGCTGTTGGAGTCTGGGGGAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCTCCTTCAGTAGCGGGTACGACATGTGCTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCGCATGCATTGCTGCTGGTAGTGCTGGTATCACTTACGACGCGAACTGGGCGAAAGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCCGTATATTACTGTGCGAGATCGGCGTTTTCGTTCGACTACGCCATGGACCTCTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGTGGAGGCGGATCTGGCGGAGGTGGTTCCGGCGGTGGCGGCTCCGGTGGAGGCGGCTCTGACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCACTTGCCAGGCCAGTCAGAGCATTAGTTCCCACTTAAACTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATAAGGCATCCACTCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTTACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGGGTTATAGTTGGGGTAATGTTGATAATGTTTTCGGCGGAGGGACCAAGGTGGAGATCAAAGGCGGTGGAGGGTCCGGCGGTGGTGGATCCCGGTCGCTGGTGGAGTCTGGGGGAGGCTTGGTCCAGCCTGGGGGGTCCCTGAGACTCTCCTGTACAGCCTCTGGATTCACCATCAGTAGCTACCACATGCAGTGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTACATCGGAACCATTAGTAGTGGTGGTAATGTATACTACGCGAGCTCCGCGAGAGGCAGATTCACCATCTCCAGACCCTCGTCCAAGAACACGGTGGATCTTCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGACTCTGGTTATAGTGATCCTATGTGGGGCCAGGGAACCCTGGTCACCGTCTCGAGCGGCGGTGGCGGTAGTGGGGGAGGCGGTTCTGGCGGCGGAGGGTCCGGCGGTGGAGGATCAGACGTTGTGATGACCCAGTCTCCATCTTCCGTGTCTGCATCTGTAGGAGACAGAGTCACCATCACCTGTCAGGCCAGTCAGAACATTAGGACTTACTTATCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGCTGCAGCCAATCTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGATTTCACTCTCACCATCAGCGACCTGGAGCCTGGCGATGCTGCAACTTACTATTGTCAGTCTACCTATCTTGGTACTGATTATGTTGGCGGTGCTTTCGGCGGAGGGACCAAGGTGGAGATCAAA
>SEQ ID 108 SI-38E33 (21D4-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)重鏈aa
AIQLTQSPSSLSASVGDRVTITCRASQGISSALA
WYQQKPGKAPKLLIYDASSLES
GVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQFNSYPFT
FGPGTKVDIKGGGGSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFSSSWIG
WVRQAPGKGLEWMGIIYPDDSDTRYSPSFQG
QVTISADKSIRTAYLQWSSLKASDTAMYYCARHVTMIWGVIIDF
WGQGTLVTVSSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTISTNAMS
WVRQAPGKGLEWIGVITGRDITYYASWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARDGGSSAITSNNI
WGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPEAAGAPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCAVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAASGFSFSSGYDMC
WVRQAPGKGLEWIACIAAGSAGITYDANWAKG
RFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSAFSFDYAMDL
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDIQMTQSPSTLSASVGDRVTITCQASQSISSHLN
WYQQKPGKAPKLLIYKASTLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQGYSWGNVDNV
FGGGTKVEIKGGGGSGGGGSRSLVESGGGLVQPGGSLRLSCTASGFTISSYHMQ
WVRQAPGKGLEYIGTISSGGNVYYASSARG
RFTISRPSSKNTVDLQMNSLRAEDTAVYYCARDSGYSDPM
WGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSDVVMTQSPSSVSASVGDRVTITCQASQNIRTYLS
WYQQKPGKAPKLLIYAAANLAS
GVPSRFSGSGSGTDFTLTISDLEPGDAATYYCQSTYLGTDYVGGA
FGGGTKVEIK
>SEQ ID 109 SI-38E33 (21D4-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)輕鏈nt
GACGTCGTGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTAGGAGACAGAGTCACCATCAATTGCCAAGCCAGTGAGAGCATTAGCAGTTGGTTAGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTATGAAGCATCCAAACTGGCATCTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAAGGCTATTTTTATTTTATTAGTCGTACTTATGTAAATTCTTTCGGCGGAGGGACCAAGGTGGAGATCAAACGTACGGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGTGT
>SEQ ID 110 SI-38E33 (21D4-LH-scFv x 284A10-Fab x PL221G5-H1L1-scFv x 466F6-H2L5-scFv)輕鏈aa
DVVMTQSPSTLSASVGDRVTINCQASESISSWLA
WYQQKPGKAPKLLIYEASKLAS
GVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQGYFYFISRTYVNS
FGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC
無
本揭示內容之前述及其他特徵將自以下描述及隨附申請專利範圍結合隨附圖式而變得更全面明顯的。應理解,該等圖式僅描繪根據本揭示內容佈置之若干實施例,且因此不視為其範疇之限制,本揭示內容將經由使用隨附圖式而描述為具有另外的特性及細節,圖式中:
第1圖展示GNC蛋白質之概括圖解,該等GNC蛋白質係藉由其多個抗原結合結構域(AgBd)及連接子之組合物來表徵;
第2圖展示GNC抗體之實例作為本文所揭示的GNC蛋白質之實施例:2A,具有EGFRvIII AgBD之四特異性GNC抗體(SI-39E18);2B,具有ROR1 AgBD之四特異性GNC抗體(SI-35E20);及2C,具有CD19 AgBD之四特異性GNC抗體(SI-38E17);
第3圖展示四特異性GNC抗體如何可經由多個AgBD結合至T細胞及腫瘤細胞;
第4圖展示結合至經人類ROR1轉染之CHO細胞的四特異性GNC抗體之實例;
第5圖展示結合至經人類4-1BB轉染之CHO細胞的四特異性GNC抗體之實例;
第6圖展示結合至經人類PD-L1轉染之CHO細胞的四特異性GNC抗體之實例;
第7圖展示示例性四特異性GNC抗體,其具有結合結構域323H7,該結合結構域323H7對B-ALL細胞系Kasumi2之ROR1介導RTCC之Ig結構域為特異性的,其中PBMC係作為效應物;
第8圖展示示例性四特異性GNC抗體,其具有結合結構域323H7,該結合結構域323H7對B-ALL細胞系Kasumi2之ROR1介導RTCC之Ig結構域為特異性的,其中CD8+、CD45RO+記憶T細胞係作為效應物;
第9圖展示示例性四特異性GNC抗體,其具有結合結構域323H7,該結合結構域323H7對B-ALL細胞系Kasumi2之ROR1介導RTCC之Ig結構域為特異性的,其中CD8+、CD45RA+初始T細胞係作為效應物;
第10圖展示示例性四特異性GNC抗體,其具有結合結構域338H4,該結合結構域338H4對B-ALL細胞系Kasumi2之ROR1介導RTCC之捲曲蛋白結構域為特異性的,其中PBMC係作為效應物;
第11圖展示示例性四特異性GNC抗體,其具有結合結構域338H4,該結合結構域338H4對B-ALL細胞系Kasumi2之ROR1介導RTCC之捲曲蛋白結構域為特異性的,其中CD8+、CD45RO+記憶T細胞係作為效應物;
第12圖展示示例性四特異性GNC抗體,其具有結合結構域338H4,該結合結構域338H4對B-ALL細胞系Kasumi2之ROR1介導RTCC之捲曲蛋白結構域為特異性的,其中CD8+、CD45RA+初始T細胞係作為效應物;
第13圖展示回應於用靶向EGFRvIII之四特異性GNC抗體進行的治療的針對膀胱癌細胞系UM-UC-3-EGFRvIII的重導引panT細胞活性;
第14圖展示量測回應於用靶向EGFRvIII之四特異性GNC抗體進行的治療的CD8 T細胞增殖之結果;
第15圖展示追蹤回應於用靶向EGFRvIII之四特異性GNC抗體進行的治療的IFNγ分泌之結果;
第16圖展示表明針對膀胱癌細胞系UM-UC-3-EGFRvIII的重導引初始T細胞細胞毒性之結果;
第17圖展示量測PBMC對用靶向EGFRvIII之四特異性GNC抗體進行的治療、CD8 T細胞之增殖的反應之結果;
第18圖展示回應於用靶向EGFRvIII之四特異性GNC抗體進行的治療的在單核細胞存在下針對膀胱癌細胞系UM-UC-3-EGFRvIII之重導引panT細胞活性之結果;
第19圖展示PD-L1及4-1BB結構域對四特異性GNC抗體之活性的功能影響及針對膀胱癌細胞系UM-UC-3-EGFRvIII之重導引PBMC細胞毒性;
第20圖展示回應於用靶向ROR1之四特異性GNC抗體進行的治療的針對Kasumi-2靶細胞系之重導引panT細胞活性之結果;
第21圖展示回應於用靶向CD19之四特異性GNC抗體進行的治療的針對Kasimu-2腫瘤細胞系之重導引PBMC活性之結果;
第22圖展示回應於用靶向CD19之四特異性GNC抗體進行的治療的CD8 T細胞增殖;及
第23圖展示回應於用靶向CD19之四特異性GNC抗體進行的治療的藉由PBMC之IFNγ產生。
國內寄存資訊 (請依寄存機構、日期、號碼順序註記)
無
國外寄存資訊 (請依寄存國家、機構、日期、號碼順序註記)
無
Claims (29)
- 一種引導及導航控制(GNC)蛋白質,其包含, 一用於一T細胞活化受體之結合結構域,一用於一腫瘤相關聯抗原之結合結構域,一用於一免疫查核點受體之結合結構域,及一用於一T細胞共刺激受體之結合結構域,其中該用於該腫瘤相關聯抗原之結合結構域不相鄰於該用於該T細胞共刺激受體之結合結構域。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中該用於該T細胞活化受體之結合結構域係相鄰於該用於該腫瘤相關聯抗原(TAA)之結合結構域。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中T細胞活化受體包含CD3。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中一免疫查核點受體包含PD-L1、PD-1、TIGIT、TIM-3、LAG-3、CTLA4、BTLA、VISTA、PDL2、CD160、LOX-1、siglec-15、CD47、或其組合。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中T細胞共刺激受體包含4-1BB、CD28、OX40、GITR、CD40L、ICOS、Light、CD27、CD30、或其組合。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中一腫瘤相關聯抗原包含ROR1、CD19、EGRFVIII、BCMA、CD20、CD33、CD123、CD22、CD30、CEA、HER2、EGFR、LMP1、LMP2A、間皮素、PSMA、EpCAM、磷脂醯肌醇蛋白聚糖-3、gpA33、GD2、TROP2、或其組合。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中該腫瘤相關聯抗原為以下各項上之一受體:一肺癌細胞、一肝癌細胞、一乳癌細胞、一結腸直腸癌細胞、一肛門癌細胞、一胰腺癌細胞、一膽囊癌細胞、一膽管癌細胞、一頭頸癌細胞、一鼻咽癌細胞、一皮膚癌細胞、一黑素瘤細胞、一卵巢癌細胞、一前列腺癌細胞、一尿道癌細胞、一肺癌細胞、一非小肺細胞癌細胞、一小細胞肺癌細胞、一腦腫瘤細胞、一神經膠質瘤細胞、一神經胚細胞瘤細胞、一食道癌細胞、一胃癌細胞、一肝癌細胞、一腎癌細胞、一膀胱癌細胞、一子宮頸癌細胞、一子宮內膜癌細胞、一甲狀腺癌細胞、一眼癌細胞、一肉瘤細胞、一骨癌細胞、一白血病細胞、一骨髓瘤細胞、一淋巴瘤細胞、或其組合。
- 如請求項1所述之引導及導航控制(GNC)蛋白質,其中該蛋白質為一包含一Fc結構域之三或四特異性抗體。
- 如請求項8所述之引導及導航控制(GNC)蛋白質,其中該T細胞活化受體包含CD3,且其中該用於CD3之結合結構域係經由一肽連接子連接至該用於該腫瘤相關聯抗原(TAA)之結合結構域以至形成一CD3-TAA對,其中該肽連接子具有不超過100個胺基酸之長度。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其中該肽連接子具有不超過20個胺基酸之一長度。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其中該肽連接子具有不超過10個胺基酸之一長度。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其中該肽連接子具有約2個胺基酸至約10個胺基酸之一長度。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其中該腫瘤相關聯抗原包含ROR1、CD19、或EGRFVIII。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其中該IgG Fc結構域中介該CD3-TAA對及該用於該免疫查核點受體之結合結構域。
- 如請求項14所述之引導及導航控制(GNC)蛋白質,其中該免疫查核點受體包含PD-L1。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其具有一N末端及一C末端,其自該N末端至該C末端串聯地包含, 該用於CD3之結合結構域,該用於EGFRVIII之結合結構域,IgG Fc結構域,該用於PD-L1之結合結構域,及該用於41-BB之結合結構域。
- 如請求項16所述之引導及導航控制(GNC)蛋白質,其包含一胺基酸序列,該胺基酸序列具有與SEQ ID NO. 80及82之一同源性百分比,其中該同源性百分比不小於98%。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其具有一N末端及一C末端,其自該N末端至該C末端串聯地包含, 該用於4-1BB之結合結構域,該用於PD-L1之結合結構域,IgG Fc結構域,該用於ROR1之結合結構域,及 該用於CD3之結合結構域。
- 如請求項18所述之引導及導航控制(GNC)蛋白質,其包含一胺基酸序列,該胺基酸序列具有與SEQ ID NO. 88及90之一同源性百分比,其中該同源性百分比不小於98%。
- 如請求項9所述之引導及導航控制(GNC)蛋白質,其具有一N末端及一C末端,其自該N末端至該C末端串聯地包含, 該用於CD3之結合結構域,該用於CD19之結合結構域,IgG Fc結構域,該用於PD-L1之結合結構域,及該用於4-1BB之結合結構域。
- 如請求項20所述之引導及導航控制(GNC)蛋白質,其包含一胺基酸序列,該胺基酸序列具有與SEQ ID NO. 104及106之一同源性百分比,其中該同源性百分比不小於98%。
- 如請求項1所述之GNC蛋白質,其其包含一胺基酸,該胺基酸具有與SEQ ID NO. 50、52、80、82、84、86、88、90、92、94、96、98、100、102、104、106、108及110之一同源性百分比的胺基酸,其中該同源性百分比不小於98%。
- 一種核酸,其編碼如請求項1所述之GNC蛋白質。
- 如請求項23所述之核酸,其具有與SEQ ID NO. 49、51、79、81、83、85、87、89、91、93、95、97、99、101、103、105、107、及109之一同源性百分比,其中該同源性百分比不小於98%。
- 一種細胞毒性細胞,其包含, 一T細胞,其具有一T細胞活化受體及一T細胞共刺激受體,及 如請求項1所述之GNC蛋白質,其經由與該T細胞活化受體、該T細胞共刺激受體、或其組合相互作用結合至該T細胞。
- 一種癌細胞,其包含 一癌細胞,其具有一腫瘤相關聯抗原,及如請求項1所述之GNC蛋白質,其經由與該腫瘤相關聯抗原相互作用結合至癌細胞。
- 一種生物學複合物,其包含, 一T細胞,其具有一T細胞活化受體及一T細胞共刺激受體,一癌細胞,其具有一腫瘤相關聯抗原,及 如請求項1所述之GNC蛋白質,其中該GNC蛋白質經由與該T細胞活化受體、該T細胞共刺激受體、或其組合相互作用結合至該T細胞,且其中該GNC蛋白質經由與該腫瘤相關聯抗原相互作用結合至該癌細胞。
- 一種醫藥組合物,其包含 如請求項1、9、16、18、或20所述之GNC蛋白質、或如請求項25所述之細胞毒性細胞、或其組合,及 一醫藥學上可接受的載劑。
- 一種一有效量的如請求項28所述之醫藥組合物在製造用於治療患有一癌症之一受試者的藥劑中之用途。
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