JP2022514815A - CDR1領域に突然変異したヒト化CD19 scFvを有するCAR-T細胞 - Google Patents
CDR1領域に突然変異したヒト化CD19 scFvを有するCAR-T細胞 Download PDFInfo
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Abstract
Description
(i)本発明の第一の側面に記載のscFv、
(ii)膜貫通ドメイン、
(iii)少なくとも一つの共刺激ドメイン、および
(iv)活性化ドメイン
を含むものを提供する。
(式中において、
各「-」は独立に連結ペプチドまたはペプチド結合である。
Lは任意にシグナルペプチド配列である。
scFvは本発明の第二の側面に記載のscFvである。
Hは任意にヒンジ領域である。
TMは膜貫通ドメインである。
Cは共刺激シグナル分子である。
CD3ζはCD3ζ由来の細胞内シグナル伝達配列である。)
(I) 本発明の第一の側面に記載のscFv、または本発明の第三の側面に記載の抗体、ならびに
(II) 任意に発現および/または精製を補助するタグ配列
を有する組み換えタンパク質を提供する。
もう一つの好適な例において、前記のタグ配列は6Hisタグを含む。
もう一つの好適な例において、前記の組み換えタンパク質(またはポリペプチド)は融合タンパク質を含む。
もう一つの好適な例において、前記の組み換えタンパク質は、単量体、二量体、または多量体である。
(a) 本発明の第一の側面に記載のscFv、または本発明の第三の側面に記載の抗体、ならびに
(b) 検出可能なマーカー、薬物、毒素、サイトカイン、放射性核種、酵素、またはこれらの組み合わせからなる群から選ばれる、前記抗体部分とカップリングするカップリング部分
を含む抗体薬物複合体を提供する。
本明細書で用いられるように、「キメラ抗原受容体(CAR)」は融合タンパク質で、抗原に結合できる細胞外ドメイン、細胞外ドメインが異なるポリペプチドから誘導される膜貫通ドメイン、および少なくとも1つの細胞内ドメインを含む。「キメラ抗原受容体(CAR)」は「キメラ受容体」、「T-body」または「キメラ免疫受容体(CIR)」と呼ばれることもある。「抗原に結合できる細胞外ドメイン」とはある抗原に結合できる任意のオリゴペプチドまたはポリペプチドである。「細胞内ドメイン」とは既知のシグナルを伝達することによって細胞内における生物過程を活性化または抑制するドメインとして作用する任意のオリゴペプチドまたはポリペプチドである。
CD19はB細胞抗原で、急性リンパ性白血病(ALL)、慢性リンパ性白血病(CLL)および非ホジキンスリンパ腫を含むすべてのB細胞悪性腫瘍において発現される。このように白血病およびリンパ腫で一般的に発現されることで、当該抗原はCAR-T細胞がターゲティングする注目の標的になっている[3]。
ヒトCD19タンパク質は95 kDaの膜貫通糖タンパク質で、556個のアミノ酸からなる。以下に示すように、20~291番目は細胞外ドメインで、292~313番目は膜貫通ドメインで、314~556番目は細胞内ドメインである(細胞外ドメインは下線で表記されている)。免疫グロブリンスーパーファミリーに属し、B細胞抗原受容体(BCR)依存性および非依存性シグナル伝達を仲介する。CD19はBCRおよびほかの細胞表面のタンパク質と結合し、ほかのキナーゼおよびリガンドとの結合によって細胞内シグナル伝達を調節する。CD19シグナルはSrcファミリーのキナーゼ、PI3Kキナーゼ、Abl、AKT依存性シグナル経路に関連する。CD19はB細胞が仲介する生存シグナル伝達および免疫応答のバイオマーカーである。
10 20 30 40 50
MPPPRLLFFL LFLTPMEVRP EEPLVVKVEE GDNAVLQCLK GTSDGPTQQL
60 70 80 90 100
TWSRESPLKP FLKLSLGLPG LGIHMRPLAI WLFIFNVSQQ MGGFYLCQPG
110 120 130 140 150
PPSEKAWQPG WTVNVEGSGE LFRWNVSDLG GLGCGLKNRS SEGPSSPSGK
160 170 180 190 200
LMSPKLYVWA KDRPEIWEGE PPCLPPRDSL NQSLSQDLTM APGSTLWLSC
210 220 230 240 250
GVPPDSVSRG PLSWTHVHPK GPKSLLSLEL KDDRPARDMW VMETGLLLPR
260 270 280 290 300
ATAQDAGKYY CHRGNLTMSF HLEITARPVL WHWLLRTGGW KVSAVTLAYL
310 320 330 340 350
IFCLCSLVGI LHLQRALVLR RKRKRMTDPT RRFFKVTPPP GSGPQNQYGN
360 370 380 390 400
VLSLPTPTSG LGRAQRWAAG LGGTAPSYGN PSSDVQADGA LGSRSPPGVG
410 420 430 440 450
PEEEEGEGYE EPDSEEDSEF YENDSNLGQD QLSQDGSGYE NPEDEPLGPE
460 470 480 490 500
DEDSFSNAES YENEDEELTQ PVARTMDFLS PHGSAWDPSR EATSLGSQSY
510 520 530 540 550
EDMRGILYAA PQLRSIRGQP GPNHEEDADS YENMDNPDGP DPAWGGGGRM
GTWSTR (配列番号16)
本発明者は、VH CDR1領域の突然変異(V27G)を含むヒト化CD19 scFvを作り、当該突然変異を含むヒト化CD19 scFvの配列に基づいて特異的にCD19を標的とするCAR-T細胞を製造した。本発明者は、CD19抗原を過剰発現する癌細胞を標的とするため、ヒト化CD19-CAR-T細胞を製造した。本発明のヒト化CD19-CAR-T細胞は白血病癌細胞に対して高レベルのIFN-γを分泌してHeLa-CD19陽性標的細胞を殺傷するが、対照のHela細胞を殺傷しない。
また、本発明は、本発明のCAR配列をコードするDNA構築物を提供する。
本発明は、本発明の発現カセットをコードするレンチウイルスベクター(LV)で形質導入された細胞を含む。形質導入されたT細胞はCARによるT細胞の応答を誘導することができる。
治療できる適応症は、CD19陽性腫瘍および過剰なB細胞による疾患を含む。CD19陽性腫瘍は、CD19陽性の非固形腫瘍(たとえば血液腫瘍、たとえば白血病やリンパ腫)または固形腫瘍を含む。本発明のCARで治療する癌の種類は、癌、胚細胞腫瘍や肉腫、および一部の白血病やリンパ性悪性腫瘍、良性腫瘍および悪性腫瘍、および悪性腫瘍、たとえば肉腫、癌やメラノーマを含むが、これらに限定されない。成人腫瘍/癌および児童腫瘍/癌も含む。
「免疫学的な有効量」、「抗腫瘍有効量」、「腫瘍抑制有効量」または「治療量」と記載する場合、施用される本発明の組成物の精確な量は、患者(対象)の年齢、体重、腫瘍の大きさ、感染または転移の程度および病症の個体差を考慮し、医師によって決められる。通常、本明細書に記載のT細胞を含む薬物組成物は、104~109個細胞/kg体重の投与量、好ましくは105~106個細胞/kg体重の投与量(これらの範囲内におけるすべての整数値を含む)で施用することができる。T細胞の組成物はこれらの投与量で数回施用してもよい。細胞は、免疫療法で公知の注入技術(たとえばRosenbergら,NewEng.J. of Med.319:1676,1988)によって施用することができる。具体的な患者対する最適な投与量および治療プランは、患者の疾患の所見をモニタリングしてそれに基づいて調整して治療することができ、医学分野の技術者によって容易に決めることができる。
対象への組成物の施用は噴霧法、注射、経口、輸液、植込みまたは移植を含む任意の便利な手段によって行ってもよい。本明細書に記載の組成物は皮下、皮内、腫瘍内、節内、脊髄内、筋肉内、静脈内(i.v.)注射または腹膜内で患者に施用されてもよい。一つの実施形態において、本発明のT細胞の組成物は皮内または皮下注射によって患者に施用される。もう一つの実施形態において、本発明のT細胞の組成物はi.v.注射によって施用することが好ましい。T細胞の組成物は直接腫瘍、リンパ節または感染の箇所に注入してもよい。
発明者は、ヒト化CD19-scFv-CAR構築物をレンチウイルスベクターのXba IとEcoR I部位にクローニングした。pCD510-FMC63-28zレンチウイルスCAR構築物は、Xba IとEco RIクローニング部位の間にヒト化CD19 scFv-41BB-CD3ζ挿入物を含む。
293T細胞にレンチウイルスが生じ、RT-PCR方法によって力価を測定した。その後、等投与量のレンチウイルスでT細胞に形質導入した。
ヒト化CD19抗体:VHとVLおよびscFv配列
発明者は、マウスCD19 FMC63 scFvクローン由来のCD19 scFvをヒト化し、そして体内治療効果のデータに基づいてCDR1に突然変異があるヒト化scFv(クローン11)を選択した。ヒト化CD19 scFvの構造は、VL-連結ペプチド-VHである。連結ペプチドはG S T S G S G K P G S G E G S T K G (配列番号7)である。
gatattcagatgacccagagcccgagcagcctgagcgcgagcgtgggcgatcgcgtgacc
attacctgccgcgcgagccaggatattagcaaatatctgaactggtatcagcagaaaccg
ggcaaagcgccgaaactgctgatttatcataccagccgcctgcatagcggcgtgccgagc
cgctttagcggcagcggcagcggcaccgattttaccctgaccattagcagcctgcagccg
gaagattttgcgacctattattgccagcagggcaacaccctgccgtatacctttggcggc
ggcaccaaagtggaaattaaa
ggctccacctctggatccggcaagcccggatctggcgagggatccaccaagggc
caggtgcagctgcaggaaagcggcccgggcctggtgaaaccgagcgaaaccctgagcctg
acctgcaccgtgagcggcggcagcctgccggattatggcgtgagctggattcgccagccg
ccgggcaaaggcctggaatggattggcgtgatttggggcagcgaaaccacctattataac
agcgcgctgaaaagccgcgtgaccattagcgtggataccagcaaaaaccagtttagcctg
aaactgagcagcgtgaccgcggcggataccgcggtgtattattgcgcgaaacattattat
tatggcggcagctatgcgatggattattggggccagggcaccctggtgaccgtgagcagc
(配列番号15)
DIQMTQSPSSLSASVGDRVTITCRASQDISKYLNWYQQKPGKAPKLLIYHTSRLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGNTLPYTFGGGTKVEIK
G S T S G S G K P G S G E G S T K G
QVQLQESGPGLVKPSETLSLTCTVSGGSLPDYGVSWIRQPPGKGLEWIGVIWGSETTYYNSALKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAKHYYYGGSYAMDYWGQGTLVTVSS(配列番号4)
ヒト化CD19-CAR配列
ヒト化CD19-CAR構築物の設計プランは図2に示す。EF1aプロモーターを持つレンチウイルスベクターでヒト化scFv CAR配列をクローニングした。
<CD8リード>
ATGGCCTTACCAGTGACCGCCTTGCTCCTGCCGCTGGCCTTGCTGCTCCACGCCGCCAGGCCG(配列番号8)
gatattcagatgacccagagcccgagcagcctgagcgcgagcgtgggcgatcgcgtgacc
attacctgccgcgcgagccaggatattagcaaatatctgaactggtatcagcagaaaccg
ggcaaagcgccgaaactgctgatttatcataccagccgcctgcatagcggcgtgccgagc
cgctttagcggcagcggcagcggcaccgattttaccctgaccattagcagcctgcagccg
gaagattttgcgacctattattgccagcagggcaacaccctgccgtatacctttggcggc
ggcaccaaagtggaaattaaa
ggctccacctctggatccggcaagcccggatctggcgagggatccaccaagggc
caggtgcagctgcaggaaagcggcccgggcctggtgaaaccgagcgaaaccctgagcctg
acctgcaccgtgagcggcggcagcctgccggattatggcgtgagctggattcgccagccg
ccgggcaaaggcctggaatggattggcgtgatttggggcagcgaaaccacctattataac
agcgcgctgaaaagccgcgtgaccattagcgtggataccagcaaaaaccagtttagcctg
aaactgagcagcgtgaccgcggcggataccgcggtgtattattgcgcgaaacattattat
tatggcggcagctatgcgatggattattggggccagggcaccctggtgaccgtgagcagc(配列番号15)
<CD8ヒンジ>
ACCACGACGCCAGCGCCGCGACCACCAACACCGGCGCCCACCATCGCGTCGCAGCCCCTGTCCCTGCGCCCAGAGGCGTGCCGGCCAGCGGCGGGGGGCGCAGTGCACACGAGGGGGCTGGACTTCGCCTGTGAT (配列番号9)
<CD8 TM>
ATCTACATCTGGGCGCCCCTGGCCGGGACTTGTGGGGTCCTTCTCCTGTCACTGGTTATCACCCTTTACTGC (配列番号10)
<4-1BB共刺激ドメイン>
AAACGGGGCAGAAAGAAACTCCTGTATATATTCAAACAACCATTTATGAGACCAGTACAAACTACTCAAGAGGAAGATGGCTGTAGCTGCCGATTTCCAGAAGAAGAAGAAGGAGGATGTGAACTG (配列番号11)
<CD3ζ>
AGAGTGAAGTTCAGCAGGAGCGCAGACGCCCCCGCGTACAAGCAGGGCCAGAACCAGCTCTATAACGAGCTCAATCTAGGACGAAGAGAGGAGTACGATGTTTTGGACAAGAGACGTGGCCGGGACCCTGAGATGGGGGGAAAGCCGAGAAGGAAGAACCCTCAGGAAGGCCTGTACAATGAACTGCAGAAAGATAAGATGGCGGAGGCCTACAGTGAGATTGGGATGAAAGGCGAGCGCCGGAGGGGCAAGGGGCACGATGGCCTTTACCAGGGTCTCAGTACAGCCACCAAGGACACCTACGACGCCCTTCACATGCAGGCCCTGCCCCCTCGCTAATAG (配列番号12)
gaattc (配列番号13)
M A L P V T A L L L P L A L L L H A A R P D I Q M T Q S P S S L S A S V G D R V T I T C R A S Q D I S K Y L N W Y Q Q K P G K A P K L L I Y H T S R L H S G V P S R F S G S G S G T D F T L T I S S L Q P E D F A T Y Y C Q Q G N T L P Y T F G G G T K V E I K G S T S G S G K P G S G E G S T K G Q V Q L Q E S G P G L V K P S E T L S L T C T V S G G S L P D Y G V S W I R Q P P G K G L E W I G V I W G S E T T Y Y N S A L K S R V T I S V D T S K N Q F S L K L S S V T A A D T A V Y Y C A K H Y Y Y G G S Y A M D Y W G Q G T L V T V S S T T T P A P R P P T P A P T I A S Q P L S L R P E A C R P A A G G A V H T R G L D F A C D I Y I W A P L A G T C G V L L L S L V I T L Y C K R G R K K L L Y I F K Q P F M R P V Q T T Q E E D G C S C R F P E E E E G G C E L R V K F S R S A D A P A Y K Q G Q N Q L Y N E L N L G R R E E Y D V L D K R R G R D P E M G G K P R R K N P Q E G L Y N E L Q K D K M A E A Y S E I G M K G E R R R G K G H D G L Y Q G L S T A T K D T Y D A L H M Q A L P P R (配列番号14)
ヒト化CD19-CAR-T細胞の白血病細胞に対する殺傷およびCD19陽性癌細胞に対するIFN-γの分泌
図2に示すように、ヒト化CD19-CAR構築物でヒト化CD19-CAR-T細胞を設計した。モックscFv(無関連scFv)でモックCAR-T細胞を製造し、陰性対照とした。FACSによってヒト化CD19-CAR-T細胞によって発現されたCD19 scFvを検出した。
ヒト化CD19-CAR-T細胞がHela-CD19細胞を殺傷したが、Hela細胞を殺傷しなかった。
ヒト化CD19 CAR-T細胞のHeLa-CD19細胞に対するIFN-γの分泌
ヒト化CD19-CAR-T細胞および標的Hela-CD19をともにインキュベートした後の上清液を収集し、そして[5]に記載のように、IFN-γの測定を行った。
ヒト化CD19-CAR-T細胞のRaji異種移植腫瘍の体内における生長に対する抑制
[4]に記載のように、Raji-ルシフェラーゼ陽性細胞をNSGマウスに静脈注射し、そして翌日に1×107個のヒト化CD19-CAR-T細胞を注射した。IVISシステムでイメージングによってRaji異種移植腫瘍細胞の生長の様子を検出した。
CD19-CAR-Tで処理されたマウスと比べ、ヒト化CD19-CAR-T細胞が顕著にRaji異種移植モデルマウスの生存時間を延長させた
Kaplan-Meier生存曲線では、マウス由来CD19-CAR-T細胞または対照モックCAR-T細胞と比べ、マウスにヒト化CD19-CAR-T細胞を注射すると、マウスの生存時間が顕著に増えたことが示された(図9)。これは、マウス由来CD19-CAR-T細胞と比べ、ヒト化CD19-CAR-T細胞が優勢で向上した治療効果を有することを示す。
突然変異していないヒト化CD19 CAR-T細胞で処理されたマウスと比べ、VH CDR1にV27G突然変異があるヒト化CD19 CARがマウスの生存時間を延長させた
VH CDR1にV27G突然変異がある(クローン11)ヒト化CD19-CARおよび突然変異がないヒト化CD19-CARを比較した。図10に示す「無突然変異huCD19」はマウスクローンFMC63由来のヒト化抗体で、CDR領域に突然変異がない。「無突然変異huCD19」はクローン11と同様のVL配列を有するが、VHのフレームワーク領域が異なる。一般的に、抗原結合活性はCDR領域に影響されるが、フレームワーク領域に影響されない。
図10における結果から、VHにV27G突然変異があるヒト化CD19 CARで治療されたマウスは、突然変異がないヒト化CD19-CAR-T細胞で治療されたマウスよりも、マウスの生存時間が延長したことが示された。
マウス体内にCD19-CAR-T細胞を注射した後、CD19またはFAB抗体およびFACSによって検出した
Raji異種移植腫瘍細胞を有するマウスにCAR-T細胞を注射した後7日目に、マウスの血液を収集して体内におけるCAR-T細胞を検出した。
1. Maus, M.V.; Haas, A.R.; Beatty, G.L.; Albelda, S.M.; Levine, B.L.; Liu, X.; Zhao, Y.; Kalos, M.; June, C.H. キメラ抗原受容体を発現するT細胞がヒトのアナフィラキシーを起こす(T cells expressing chimeric antigen receptors can cause anaphylaxis in humans). Cancer Immunol Res 2013, 1, 26-31.
2. Maus, M.V.; Grupp, S.A.; Porter, D.L.; June, C.H. 抗体で修飾されたT細胞:CARが血液系悪性腫瘍において重要な役割を果たす(Antibody-modified t cells: Cars take the front seat for hematologic malignancies). Blood 2014, 123, 2625-2635.
3. Eshhar, Z.; Waks, T.; Gross, G. T体/CAR T細胞の出現(The emergence of t-bodies/car t cells). Cancer J 2014, 20, 123-126.
4. Berahovich, R.; Xu, S.; Zhou, H.; Harto, H.; Xu, Q.; Garcia, A.; Liu, F.; Golubovskaya, V.M.; Wu, L. Flagで標識されたCD19特異性のCAR-T細胞が体内と体外でCD19を持つ固形腫瘍細胞を除く(Flag-tagged cd19-specific car-t cells eliminate cd19-bearing solid tumor cells in vitro and in vivo). Front Biosci (Landmark Ed) 2017, 22, 1644-1654.
5. Golubovskaya, V.; Berahovich, R.; Zhou, H.; Xu, S.; Harto, H.; Li, L.; Chao, C.C.; Mao, M.M.; Wu, L. CD47-CAR-T細胞が有効に標的癌細胞を殺傷しかつ膵臓腫瘍の生長を遮断する(Cd47-car-t cells effectively kill target cancer cells and block pancreatic tumor
growth). Cancers (Basel) 2017, 9.
Claims (10)
- ヒト化CD19の一本鎖可変断片(scFv)であって、配列番号6で示されるアミノ酸配列を有するVHおよび配列番号5で示されるアミノ酸配列を有するVLを含むこと、
を特徴とするscFv。 - さらにVHとVLの間のリンカーを含むこと、
を特徴とする請求項1に記載のscFv。 - 配列番号7で示されるアミノ酸配列を有すること、
を特徴とする請求項2に記載のscFv。 - キメラ抗原受容体(CAR)融合タンパク質であって、N末端からC末端へ、
(i)請求項1に記載のscFv、
(ii)膜貫通ドメイン、
(iii)少なくとも一つの共刺激ドメイン、および
(iv)活性化ドメイン
を含むことを特徴とするCAR融合タンパク質。 - 配列番号14で示されるアミノ酸配列を有すること、
を特徴とする請求項4に記載のCAR融合タンパク質。 - 抗ヒトCD19の抗体であって、配列番号6で示されるアミノ酸配列を有するVHおよび配列番号5で示されるアミノ酸配列を有するVLを含むこと、
を特徴とする抗体。 - 請求項1に記載のscFv、または請求項4に記載のCAR融合タンパク質、または請求項6に記載の抗体をコードすることを特徴とする核酸分子。
- 請求項7に記載の核酸分子を含むこと、
を特徴とするベクター。 - 請求項8に記載のベクターまたは外来染色体として組み込まれた請求項7に記載の核酸分子を含むか、染色体に外来の請求項7に記載の核酸分子が組み込まれているか、あるいは請求項1に記載のscFv、または請求項4に記載のCAR融合タンパク質、または請求項6に記載の抗体を発現すること、
を特徴とする宿主細胞。 - 請求項1に記載のscFv、または請求項4に記載のCAR融合タンパク質、または請求項8に記載のベクター、または請求項9に記載の細胞と、薬学的に許容される担体、希釈剤または賦形剤とを含有すること、
を特徴とする製剤。
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Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107226867A (zh) * | 2017-07-25 | 2017-10-03 | 重庆精准生物技术有限公司 | 抗人cd19抗原的嵌合抗原受体及其应用 |
CN107312091A (zh) * | 2017-05-02 | 2017-11-03 | 重庆精准生物技术有限公司 | 靶向人cd19抗原的人源化单克隆抗体 |
CN107383196A (zh) * | 2017-08-30 | 2017-11-24 | 广州百暨基因科技有限公司 | 人源化抗cd19的抗原结合片段 |
US20180153977A1 (en) * | 2015-07-24 | 2018-06-07 | Innovative Cellular Therapeutics CO., LTD. | Humanized anti-cd19 antibody and use thereof with chimeric antigen receptor |
WO2018200496A1 (en) * | 2017-04-24 | 2018-11-01 | Kite Pharma, Inc. | Humanized antigen-binding domains against cd19 and methods of use |
Family Cites Families (7)
Publication number | Priority date | Publication date | Assignee | Title |
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TWI654206B (zh) * | 2013-03-16 | 2019-03-21 | 諾華公司 | 使用人類化抗-cd19嵌合抗原受體治療癌症 |
WO2017172952A1 (en) * | 2016-04-01 | 2017-10-05 | Promab Biotechnologies, Inc. | Flag tagged cd19-car-t cells |
CN117024588A (zh) * | 2016-09-06 | 2023-11-10 | 华道(上海)生物医药有限公司 | 抗cd19抗体在制备白血病治疗药物中的用途 |
CN108330133B (zh) * | 2017-01-20 | 2020-06-30 | 上海恒润达生生物科技有限公司 | 靶向cd19嵌合抗原受体并对其双重修饰的方法及其用途 |
CN107880128B (zh) * | 2017-12-21 | 2021-03-02 | 常州费洛斯药业科技有限公司 | 一种抗cd19的全人源抗体或抗体片段及其方法和应用 |
CN109053899B (zh) * | 2017-12-22 | 2021-11-16 | 湖南远泰生物技术有限公司 | 一种含人转铁蛋白抗原表位序列的嵌合体抗原受体 |
CN108864307A (zh) * | 2018-07-23 | 2018-11-23 | 北京多赢时代科技有限公司 | 信号肽优化靶向cd19的嵌合抗原受体、表达该嵌合抗原受体的t细胞及制备方法和应用 |
-
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Patent Citations (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20180153977A1 (en) * | 2015-07-24 | 2018-06-07 | Innovative Cellular Therapeutics CO., LTD. | Humanized anti-cd19 antibody and use thereof with chimeric antigen receptor |
WO2018200496A1 (en) * | 2017-04-24 | 2018-11-01 | Kite Pharma, Inc. | Humanized antigen-binding domains against cd19 and methods of use |
CN107312091A (zh) * | 2017-05-02 | 2017-11-03 | 重庆精准生物技术有限公司 | 靶向人cd19抗原的人源化单克隆抗体 |
CN107226867A (zh) * | 2017-07-25 | 2017-10-03 | 重庆精准生物技术有限公司 | 抗人cd19抗原的嵌合抗原受体及其应用 |
CN107383196A (zh) * | 2017-08-30 | 2017-11-24 | 广州百暨基因科技有限公司 | 人源化抗cd19的抗原结合片段 |
Non-Patent Citations (1)
Title |
---|
CELLULAR IMMUNOLOGY, vol. Vol. 304-305, JPN6022029552, 2016, pages 49 - 54, ISSN: 0004828325 * |
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