TW202233248A - Combination therapy - Google Patents

Combination therapy Download PDF

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TW202233248A
TW202233248A TW110141303A TW110141303A TW202233248A TW 202233248 A TW202233248 A TW 202233248A TW 110141303 A TW110141303 A TW 110141303A TW 110141303 A TW110141303 A TW 110141303A TW 202233248 A TW202233248 A TW 202233248A
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antibody
seq
amino acid
acid sequence
heavy chain
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TW110141303A
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席拉 佳黛
艾利森 史密斯
凱立 克魯斯曼
伯納德 亞瑟 劉
艾普斯 希瑟 凡
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美商西健公司
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Abstract

Provided herein are methods of treating cancer with an antibody that binds an immune cell engager in combination with an antibody-drug conjugate.

Description

組合療法combination therapy

本發明提供藉由結合免疫細胞接合子之抗體與抗體-藥物結合物的組合治療癌症之方法。The present invention provides methods of treating cancer by combining antibodies that bind to immune cell engagers and antibody-drug conjugates.

免疫腫瘤學治療劑及抗體-藥物結合物(ADC)已用於治療患者之癌症。沒有一類治療劑已經能夠治療目標適應症之全部患者。本發明解決此問題及其他問題。Immuno-oncology therapeutics and antibody-drug conjugates (ADCs) have been used to treat cancer in patients. No class of therapeutic agents has been able to treat all patients of a target indication. The present invention addresses this and other problems.

實施例1.一種治療癌症之方法，其包含向患有癌症之個體投與：(1)抗體-藥物結合物(ADC)，其包含結合腫瘤相關抗原之第一抗體及細胞毒性劑，其中細胞毒性劑為微管蛋白干擾劑；及(2)第二抗體，其結合於免疫細胞接合子，其中第二抗體包含增強與一或多種活化FcγR之結合的Fc，其中活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。Embodiment 1. A method of treating cancer, comprising administering to an individual with cancer: (1) an antibody-drug conjugate (ADC) comprising a primary antibody that binds a tumor-associated antigen and a cytotoxic agent, wherein cells The toxic agent is a tubulin interfering agent; and (2) a second antibody that binds to an immune cell engager, wherein the second antibody comprises an Fc that enhances binding to one or more activated FcγRs, wherein the activated FcγRs include FcγRIIIa, FcγRIIa, and /or one or more of FcyRI.

實施例2.如實施例1之方法，其中第二抗體包含增強與至少FcγRIIIa之結合的Fc。Embodiment 2. The method of embodiment 1, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa.

實施例3.如實施例1之方法，其中第二抗體包含增強與至少FcγRIIIa及FcγRIIa之結合的Fc。Embodiment 3. The method of embodiment 1, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRIIa.

實施例4.如實施例1之方法，其中第二抗體包含增強與至少FcγRIIIa及FcγRI之結合的Fc。Embodiment 4. The method of embodiment 1, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRI.

實施例5.如實施例1之方法，其中第二抗體包含增強與FcγRIIIa、FcγRIIa及FcγRI之結合的Fc。Embodiment 5. The method of embodiment 1, wherein the second antibody comprises an Fc that enhances binding to FcyRIIIa, FcyRIIa, and FcyRI.

實施例6.如實施例1至5中任一項之方法，其中第二抗體之Fc減弱與一或多種抑制性FcγR之結合。Embodiment 6. The method of any one of embodiments 1-5, wherein the Fc of the second antibody attenuates binding to one or more inhibitory FcyRs.

實施例7.如實施例6之方法，其中第二抗體之Fc減弱與FcγRIIb之結合。Embodiment 7. The method of embodiment 6, wherein the Fc of the second antibody attenuates binding to FcyRIIb.

實施例8.如實施例1至7中任一項之方法，其中第二抗體之Fc具有減少之岩藻糖含量及/或已經工程改造以包含一或多個突變，使得Fc增強與一或多種活化FcγR之結合。Embodiment 8. The method of any one of embodiments 1 to 7, wherein the Fc of the second antibody has reduced fucose content and/or has been engineered to contain one or more mutations such that Fc enhancement is associated with one or more of the mutations. Binding of various activated FcγRs.

實施例9.如實施例8之方法，其中第二抗體未經岩藻糖基化。Embodiment 9. The method of embodiment 8, wherein the second antibody is not fucosylated.

實施例10.如實施例8之方法，其中第二抗體包含重鏈恆定區中之取代S293D、A330L及I332E。Embodiment 10. The method of embodiment 8, wherein the second antibody comprises the substitutions S293D, A330L and I332E in the heavy chain constant region.

實施例11.一種治療癌症之方法，其包含向患有癌症之個體投與抗體-藥物結合物，其中抗體-藥物結合物包含：第一抗體，其與細胞毒性劑結合，其中細胞毒性劑為微管蛋白干擾劑；及第二抗體，其結合免疫細胞接合子，其中第二抗體未經岩藻糖基化。Embodiment 11. A method of treating cancer, comprising administering to an individual with cancer an antibody-drug conjugate, wherein the antibody-drug conjugate comprises: a first antibody bound to a cytotoxic agent, wherein the cytotoxic agent is a tubulin interfering agent; and a second antibody that binds to an immune cell conjugator, wherein the second antibody is not fucosylated.

實施例12.如實施例1至11中任一項之方法，其中第一抗體結合腫瘤相關抗原。Embodiment 12. The method of any one of embodiments 1-11, wherein the first antibody binds to a tumor-associated antigen.

實施例13.一種治療癌症之方法，其包含向患有癌症之個體投與：(1)抗體-藥物結合物(ADC)，其中ADC包含結合腫瘤相關抗原之第一抗體及細胞毒性劑，其中細胞毒性劑為微管蛋白干擾劑；及(2)第二抗體，其結合免疫細胞接合子，其中第二抗體包含相對於相同同種型之對應野生型Fc具有增強ADCC活性之Fc。Embodiment 13. A method of treating cancer, comprising administering to an individual with cancer: (1) an antibody-drug conjugate (ADC), wherein the ADC comprises a first antibody that binds a tumor-associated antigen and a cytotoxic agent, wherein The cytotoxic agent is a tubulin interfering agent; and (2) a secondary antibody that binds an immune cell engager, wherein the secondary antibody comprises an Fc with enhanced ADCC activity relative to a corresponding wild-type Fc of the same isotype.

實施例14.如實施例13之方法，其中第二抗體包含相對於相同同種型之對應野生型Fc具有增強ADCC及ADCP活性的Fc。Embodiment 14. The method of embodiment 13, wherein the second antibody comprises an Fc having enhanced ADCC and ADCP activity relative to a corresponding wild-type Fc of the same isotype.

實施例15.如實施例13或14之方法，其中第二抗體未經岩藻糖基化。Embodiment 15. The method of embodiment 13 or 14, wherein the second antibody is not fucosylated.

實施例16.如實施例13至15中任一項之方法，其中第二抗體包含增強與一或多種活化FcγR之結合的Fc，其中活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。Embodiment 16. The method of any one of embodiments 13-15, wherein the second antibody comprises an Fc that enhances binding to one or more activated FcyRs, wherein the activated FcyRs comprise one or more of FcyRIIIa, FcyRIIa, and/or FcyRI By.

實施例17.如實施例16之方法，其中第二抗體包含增強與至少FcγRIIIa之結合的Fc。Embodiment 17. The method of embodiment 16, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa.

實施例18.如實施例16之方法，其中第二抗體包含增強與至少FcγRIIIa及FcγRIIa之結合的Fc。Embodiment 18. The method of embodiment 16, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRIIa.

實施例19.如實施例16之方法，其中第二抗體包含增強與至少FcγRIIIa及FcγRI之結合的Fc。Embodiment 19. The method of embodiment 16, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRI.

實施例20.如實施例16之方法，其中第二抗體包含增強與FcγRIIIa、FcγRIIa及FcγRI之結合的Fc。Embodiment 20. The method of embodiment 16, wherein the second antibody comprises an Fc that enhances binding to FcyRIIIa, FcyRIIa, and FcyRI.

實施例21.如實施例13至20中任一項之方法，其中第二抗體之Fc減弱與一或多種抑制性FcγR之結合。Embodiment 21. The method of any one of embodiments 13-20, wherein the Fc of the second antibody attenuates binding to one or more inhibitory FcyRs.

實施例22.如實施例21之方法，其中第二抗體之Fc減弱與FcγRIIb之結合。Embodiment 22. The method of embodiment 21, wherein the Fc of the second antibody attenuates binding to FcyRIIb.

實施例23.如實施例1至22中任一項之方法，其中第一抗體結合選自以下之抗原：5T4 (TPBG)、ADAM-9 、AG-7、ALK、ALP、AMHRII、APLP2、ASCT2、AVB6、AXL (UFO)、B7-H3 (CD276)、B7-H4、BCMA、C3a、C3b、C4.4a (LYPD3)、C5、C5a、CA6、CA9、CanAg、碳酸酐酶IX (CAIX)、組織蛋白酶D、CCR7、CD1、CD10、CD100、CD101、CD102、CD103、CD104、CD105、CD106、CD107a、CD107b、CD108、CD109、CD111、CD112、CD113、CD116、CD117、CD118、CD119、CD11A、CD11b、CD11c、CD120a、CD121a、CD121b、CD122、CD123、CD124、CD125、CD126、CD127、CD13、CD130、CD131、CD132、CD133、CD135、CD136、CD137、CD138、CD14、CD140a、CD140b、CD141、CD142、CD143、CD144、CD146、CD147、CD148、CD15、CD150、CD151、CD154、CD155、CD156a、CD156b、CD156c、CD157、CD158b2、CD158e、CD158f1、CD158h、CD158i、CD159a、CD16、CD160、CD161、CD162、CD163、CD164、CD166、CD167b、CD169、CD16a、CD16b、CD170、CD171、CD172a、CD172b、CD172g、CD18、CD180、CD181、CD183、CD184、CD185、CD19、CD194、CD197、CD1a、CD1b、CD1c、CD1d、CD2、CD20、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD208、CD21、CD213a1、CD213a2、CD217、CD218a、CD22、CD220、CD221、CD222、CD224、CD226、CD228、CD229、CD23、CD230、CD232、CD239、CD243、CD244、CD248、CD249、CD25、CD26、CD265、CD267、CD269、CD27、CD272、CD273、CD274、CD275、CD279、CD28、CD280、CD281、CD282、CD283、CD284、CD289、CD29、CD294、CD295、CD298、CD3、CD3 ε、CD30、CD300f、CD302、CD304、CD305、CD307、CD31、CD312、CD315、CD316、CD317、CD318、CD319、CD32、CD321、CD322、CD324、CD325、CD326、CD327、CD328、CD32b、CD33、CD331、CD332、CD333、CD334、CD337、CD339、CD34、CD340、CD344、CD35、CD352、CD36、CD37、CD38、CD39、CD3d、CD3g、CD4、CD41、CD42d、CD44、CD44v6、CD45、CD46、CD47、CD48、CD49a、CD49b、CD49c、CD49d、CD49e、CD49f、CD5、CD50、CD51、CD51 (整合素α-V)、CD52、CD53、CD54、CD55、CD56、CD58、CD59、CD6、CD61、CD62L、CD62P、CD63、CD64、CD66a-e、CD67、CD68、CD69、CD7、CD70、CD70L、CD71、CD71 (TfR)、CD72、CD73、CD74、CD79a、CD79b、CD8、CD80、CD82、CD83、CD84、CD85f、CD85i、CD85j、CD86、CD87、CD89、CD90、CD91、CD92、CD95、CD96、CD97、CD98、CDH6、CDH6 (鈣黏素6)、CDw210a、CDw210b、CEA、CEACAM5、CEACAM6、CFC1B、cKIT、CLDN18.2 (密連蛋白18.2)、CLDN6、CLDN9、CLL-1、c-MET、補體因子C3、Cripto、CSP-1、CXCR5、DCLK1、DLK-1、DLL3、DPEP3、DR5 (死亡受體5)、抗黏附素(Dysadherin)、EFNA4、EGFR、EGFR野生型、EGFRviii、EGP-1 (TROP-2)、EGP-2、EMP2、ENPP3、EpCAM、EphA2、EphA3、Ephrin-A4 (EFNA4)、ETBR、FAP、FcRH5、FGFR2、FGFR3、FLT3、FOLR、FOLR1、FOLR-α、FSH、GCC、GD2、GD3、globo H、GPC1、GPC-1、GPC3、GPNMB、GPR20、HER2、HER-2、HER3、HER-3、HGFR (c-Met)、HLA-DR、HM1.24、HSP90、Ia、IGF-1R、IL-13R、IL-15、IL1RAP、IL-2、IL-3、IL-4、IL7R、整合素αVβ3 (integrin alphaVbeta3/integrin αVβ3)、整合素β-6、介白素-4受體(IL4R)、KAAG-1、KLK2、LAMP-1、Le(y)、路易斯Y抗原(Lewis Y antigen)、LGALS3BP、LGR5、LH/hCG、LHRH、脂筏、LIV-1 (SLC39A6或ZIP6)、LRP-1、LRRC15、LY6E、巨噬細胞甘露糖受體1、MAGE、間皮素(MSLN)、MET、I類MHC鏈相關蛋白A及B (MICA及MICB)、MN/CA IX、MRC2、MT1-MMP、MTX3、MTX5、MUC1、MUC16、MUC2、MUC3、MUC4、MUC5、MUC5ac、NaPi2b、NCA-90、NCA-95、連接素-4、Notch3、核仁素、OAcGD2、OT-MUC1 (腫瘤栓繫MUC1)、OX001L、P1GF、PAM4抗原、p-鈣黏素(鈣黏素3)、PD-L1、磷脂醯基絲胺酸(PS)、PRLR、促乳素受體(PRLR)、假單胞菌(Pseudomonas)、PSMA、PTK4、PTK7、受體酪胺酸激酶(RTK)、RNF43、ROR1、ROR2、SAIL、SEZ6、SLAMF7、SLC44A4、SLITRK6、SLMAMF7 (CS1)、SLTRK6、分選蛋白(Sortilin) (SORT1)、SSEA-4、SSTR2、金黃色葡萄球菌(Staphylococcus aureus) (抗生素劑)、STEAP-1、STING、STn、T101、TAA、TAC、TDGF1、肌腱蛋白、TENB2、TGF-B、Thomson-Friedenreich抗原、Thy1.1、TIM-1、組織因子(TF；CD142)、TM4SF1、Tn抗原、TNF-alpha (TNFα)、TRA-1-60、TRAIL受體(R1及R2)、TROP-2、腫瘤相關糖蛋白72 (TAG-72)、uPAR、VEGFR、VEGFR-2及xCT。Embodiment 23. The method of any one of embodiments 1 to 22, wherein the first antibody binds to an antigen selected from the group consisting of: 5T4 (TPBG), ADAM-9, AG-7, ALK, ALP, AMHRII, APLP2, ASCT2 , AVB6, AXL (UFO), B7-H3 (CD276), B7-H4, BCMA, C3a, C3b, C4.4a (LYPD3), C5, C5a, CA6, CA9, CanAg, Carbonic Anhydrase IX (CAIX), Cathepsin D, CCR7, CD1, CD10, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107a, CD107b, CD108, CD109, CD111, CD112, CD113, CD116, CD117, CD118, CD119, CD11A, CD11b, CD11c, CD120a, CD121a, CD121b, CD122, CD123, CD124, CD125, CD126, CD127, CD13, CD130, CD131, CD132, CD133, CD135, CD136, CD137, CD138, CD14, CD140a, CD140b, CD141, CD142, CD14 CD144, CD146, CD147, CD148, CD15, CD150, CD151, CD154, CD155, CD156a, CD156b, CD156c, CD157, CD158b2, CD158e, CD158f1, CD158h, CD158i, CD159a, CD16, CD160, CD161, CD162, CD163 CD166, CD167b, CD169, CD16a, CD16b, CD170, CD171, CD172a, CD172b, CD172g, CD18, CD180, CD181, CD183, CD184, CD185, CD19, CD194, CD197, CD1a, CD1b, CD1c, CD1d, CD2, CD20, CD200, CD201, CD202b, CD203c, CD204, CD205, CD206, CD208, CD21, CD213a1, CD213a2, CD217, CD218a, CD22, CD220, CD221, CD222, CD224, CD226, CD228, CD229, CD23, CD230, CD232, CD239 CD243, CD244, CD248, CD249, CD25, CD26, CD265, CD267, CD269, CD27, CD2 72, CD273, CD274, CD275, CD279, CD28, CD280, CD281, CD282, CD283, CD284, CD289, CD29, CD294, CD295, CD298, CD3, CD3ε, CD30, CD300f, CD302, CD304, CD305, CD307, CD31 , CD312, CD315, CD316, CD317, CD318, CD319, CD32, CD321, CD322, CD324, CD325, CD326, CD327, CD328, CD32b, CD33, CD331, CD332, CD333, CD334, CD337, CD339, CD34, CD340, CD344 , CD35, CD352, CD36, CD37, CD38, CD39, CD3d, CD3g, CD4, CD41, CD42d, CD44, CD44v6, CD45, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD5, CD50 , CD51, CD51 (integrin alpha-V), CD52, CD53, CD54, CD55, CD56, CD58, CD59, CD6, CD61, CD62L, CD62P, CD63, CD64, CD66a-e, CD67, CD68, CD69, CD7, CD70, CD70L, CD71, CD71 (TfR), CD72, CD73, CD74, CD79a, CD79b, CD8, CD80, CD82, CD83, CD84, CD85f, CD85i, CD85j, CD86, CD87, CD89, CD90, CD91, CD92, CD95 , CD96, CD97, CD98, CDH6, CDH6 (Cadherin 6), CDw210a, CDw210b, CEA, CEACAM5, CEACAM6, CFC1B, cKIT, CLDN18.2 (claudin 18.2), CLDN6, CLDN9, CLL-1, c -MET, complement factor C3, Cripto, CSP-1, CXCR5, DCLK1, DLK-1, DLL3, DPEP3, DR5 (Death Receptor 5), Dysadherin, EFNA4, EGFR, EGFR wild type, EGFRviii, EGP-1 (TROP-2), EGP-2, EMP2, ENPP3, EpCAM, EphA2, EphA3, Ephrin-A4 (EFNA4), ETBR, FAP, FcRH5, FGFR2, FGFR3, FLT3, FOLR, F OLR1, FOLR-α, FSH, GCC, GD2, GD3, globo H, GPC1, GPC-1, GPC3, GPNMB, GPR20, HER2, HER-2, HER3, HER-3, HGFR (c-Met), HLA- DR, HM1.24, HSP90, Ia, IGF-1R, IL-13R, IL-15, IL1RAP, IL-2, IL-3, IL-4, IL7R, integrin alphaVbeta3/integrin alphaVbeta3, integration β-6, interleukin-4 receptor (IL4R), KAAG-1, KLK2, LAMP-1, Le(y), Lewis Y antigen, LGALS3BP, LGR5, LH/hCG, LHRH, Lipid rafts, LIV-1 (SLC39A6 or ZIP6), LRP-1, LRRC15, LY6E, macrophage mannose receptor 1, MAGE, mesothelin (MSLN), MET, MHC class I chain-associated proteins A and B ( MICA and MICB), MN/CA IX, MRC2, MT1-MMP, MTX3, MTX5, MUC1, MUC16, MUC2, MUC3, MUC4, MUC5, MUC5ac, NaPi2b, NCA-90, NCA-95, Connexin-4, Notch3 , nucleolin, OAcGD2, OT-MUC1 (tumor tethered MUC1), OX001L, P1GF, PAM4 antigen, p-cadherin (cadherin 3), PD-L1, phosphatidylserine (PS), PRLR, prolactin receptor (PRLR), Pseudomonas, PSMA, PTK4, PTK7, receptor tyrosine kinase (RTK), RNF43, ROR1, ROR2, SAIL, SEZ6, SLAMF7, SLC44A4, SLITRK6 , SLMAMF7 (CS1), SLTRK6, Sortilin (SORT1), SSEA-4, SSTR2, Staphylococcus aureus (antibiotic agent), STEAP-1, STING, STn, T101, TAA, TAC , TDGF1, tenascin, TENB2, TGF-B, Thomson-Friedenreich antigen, Thy1.1, TIM-1, tissue factor (TF; CD142), TM4SF1, Tn antigen, TNF-alpha (TNFα), TRA-1-60 , TRAIL receptors (R1 and R2), TROP-2, tumor-associated glycoprotein 72 (TAG-72), uPAR, VEGFR, VEGFR-2 and xCT.

實施例24.如實施例1至23中任一項之方法，其中第一抗體不結合連接素-4。Embodiment 24. The method of any one of embodiments 1-23, wherein the first antibody does not bind connexin-4.

實施例25.如實施例1至24中任一項之方法，其中方法不包含投與抗體-藥物結合物，該抗體-藥物結合物包含結合連接素-4之抗體。Embodiment 25. The method of any one of embodiments 1-24, wherein the method does not comprise administering an antibody-drug conjugate comprising an antibody that binds connexin-4.

實施例26.如實施例1至25中任一項之方法，其中第一抗體結合選自以下之抗原：CD71、Axl、AMHRII及LGR5、Axl、CA9、CD142、CD20、CD22、CD228、CD248、CD30、CD33、CD37、CD48、CD7、CD71、CD79b、CLDN18.2、CLDN6、c-MET、EGFR、EphA2、ETBR、FCRH5、GCC、Globo H、gpNMB、HER-2、IL7R、整合素β-6、KAAG-1、LGR5、LIV-1、LRRC15、Ly6E、間皮素(MSLN)、MET、MRC2、MUC16、NaPi2b、連接素-4、OT-MUC1 (腫瘤栓繫-MUC1)、PSMA、ROR1、SLAMF7、SLC44A4、SLITRK6、STEAP-1、STn、TIM-1、TRA-1-60及腫瘤相關糖蛋白72 (TAG-72)。Embodiment 26. The method of any one of embodiments 1 to 25, wherein the first antibody binds to an antigen selected from the group consisting of CD71, Axl, AMHRII and LGR5, Axl, CA9, CD142, CD20, CD22, CD228, CD248, CD30, CD33, CD37, CD48, CD7, CD71, CD79b, CLDN18.2, CLDN6, c-MET, EGFR, EphA2, ETBR, FCRH5, GCC, Globo H, gpNMB, HER-2, IL7R, integrin beta-6 , KAAG-1, LGR5, LIV-1, LRRC15, Ly6E, Mesothelin (MSLN), MET, MRC2, MUC16, NaPi2b, Connexin-4, OT-MUC1 (Tumor Tethered-MUC1), PSMA, ROR1, SLAMF7, SLC44A4, SLITRK6, STEAP-1, STn, TIM-1, TRA-1-60 and tumor-associated glycoprotein 72 (TAG-72).

實施例27.如實施例1至25中任一項之方法，其中第一抗體結合選自以下之抗原：BCMA、GPC1、CD30、cMET、SAIL、HER3、CD70、CD46、CD48、HER2、5T4、ENPP3、CD19、EGFR及EphA2。Embodiment 27. The method of any one of embodiments 1 to 25, wherein the first antibody binds to an antigen selected from the group consisting of BCMA, GPC1, CD30, cMET, SAIL, HER3, CD70, CD46, CD48, HER2, 5T4, ENPP3, CD19, EGFR and EphA2.

實施例28.如實施例1至25中任一項之方法，其中第一抗體結合選自以下之抗原：Her2、TROP2、BCMA、cMet、整合素αVβ6 (integrin alphVbeta6/integrin αVβ6)、CD22、CD79b、CD30、CD19、CD70、CD228、CD47及CD48。Embodiment 28. The method of any one of embodiments 1 to 25, wherein the primary antibody binds to an antigen selected from the group consisting of Her2, TROP2, BCMA, cMet, integrin alphVbeta6/integrin αVβ6, CD22, CD79b , CD30, CD19, CD70, CD228, CD47 and CD48.

實施例29.如實施例1至25中任一項之方法，其中第一抗體結合選自以下之抗原：CD142、整合素β-6、整合素αVβ6、ENPP3、CD19、Ly6E、cMET、C4.4a、CD37、MUC16、STEAP-1、LRRC15、SLITRK6、ETBR、FCRH5、Axl、EGFR、CD79b、BCMA、CD70、PSMA、CD79b、CD228、CD48、LIV-1、EphA2、SLC44A4、CD30及sTn。Embodiment 29. The method of any one of embodiments 1 to 25, wherein the first antibody binds to an antigen selected from the group consisting of CD142, integrin β-6, integrin αVβ6, ENPP3, CD19, Ly6E, cMET, C4. 4a, CD37, MUC16, STEAP-1, LRRC15, SLITRK6, ETBR, FCRH5, Axl, EGFR, CD79b, BCMA, CD70, PSMA, CD79b, CD228, CD48, LIV-1, EphA2, SLC44A4, CD30 and sTn.

實施例30.如實施例1至26中任一項之方法，其中微管蛋白干擾劑為奧瑞他汀(auristatins)、微管蛋白裂解素(tubulysin)、秋水仙鹼(colchicine)、長春花生物鹼(vinca alkaloid)、紫杉烷(taxane)、克瑞托欣(cryptophycin)、類美登素(maytansinoid)或哈米特林(hemiasterlin)。Embodiment 30. The method of any one of embodiments 1 to 26, wherein the tubulin interfering agent is auristatins, tubulysin, colchicine, vinca biological Vinca alkaloid, taxane, cryptophycin, maytansinoid or hemiasterlin.

實施例31.如實施例30之方法，其中微管蛋白干擾劑為奧瑞他汀。Embodiment 31. The method of embodiment 30, wherein the tubulin interfering agent is auristatin.

實施例32.如實施例1至31中任一項之方法，其中微管蛋白干擾劑為尾海兔素(dolostatin)-10、MMAE (N-甲基纈胺酸-纈胺酸-多拉索因(dolaisoleuine)-多拉普因(dolaproine)-降麻黃鹼)、MMAF (N-甲基纈胺酸-纈胺酸-多拉索因-多拉普因-苯丙胺酸)、奧瑞他汀F、AEB、AEVB或AFP (奧瑞他汀苯丙胺酸苯二胺)。Embodiment 32. The method of any one of embodiments 1 to 31, wherein the tubulin interfering agent is dolostatin-10, MMAE (N-methylvaline-valine-dola Dolaisoleuine-dolaproine-noephedrine), MMAF (N-methylvaline-valine-dolasoine-dolaproine-phenylalanine), orrelline Statin F, AEB, AEVB or AFP (auristatin phenylalanine phenylenediamine).

實施例33.如實施例1至32中任一項之方法，其中微管蛋白干擾劑為MMAE。Embodiment 33. The method of any one of embodiments 1-32, wherein the tubulin interfering agent is MMAE.

實施例33-1.如實施例33之方法，其中抗體-藥物結合物包含MMAE且係選自：DP303c，亦稱為SYSA1501，其靶向HER-2 (CSPC Pharmaceutical；Dophen Biomed)；SIA01-ADC，亦稱為ST1，其靶向STn (Siamab Therapeutics)；拉地妥珠單抗維多汀(Ladiratuzumab vedotin)，亦稱為SGN-LIV1A，其靶向LIV-1 (Merck & Co., Inc.；Seagen (Seattle Genetics) Inc.)；ABBV-085，亦稱為沙馬妥單抗維多汀(Samrotamab vedotin)，其靶向LRRC15 (Abbvie; Seagen (Seattle Genetics) Inc.)；DMOT4039A，亦稱為RG7600；αMSLN-MMAE，其靶向間皮素(MSLN) (Roche-Genentech)；RC68，亦稱為Remegen EGFR ADC，其靶向EGFR (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.))；RC108，亦稱為RC108-ADC，其靶向c-MET (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.))；CMG901，亦稱為MRG005，其靶向CLDN18.2 (Keymed Biosciences；Lepu biotech；Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd))；YBL-001，亦稱為LCB67，其靶向DLK-1 (Lego Chem Biosciences；Pyxis Oncology；Y-Biologics)；DCDS0780A，亦稱為艾妥珠單抗維多汀(Iladatuzumab vedotin)；RG7986，其靶向CD79b (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；替索圖單抗維多汀(Tisotumab vedotin)，亦稱為Humax-TF-ADC；tf-011-mmae；TIVDAK™，其靶向CD142 (GenMab; Seagen (Seattle Genetics) Inc.)；GO-3D1-ADC，亦稱為humAb-3D1-MMAE ADC，其靶向MUC1-C (Genus Oncology LLC)；ALT-P7，亦稱為HM2-MMAE，其靶向HER-2 (Alteogen, Inc.；Levena Biopharma；3SBio, Inc.)；凡多土珠單抗維多汀(Vandortuzumab vedotin)，亦稱為DSTP3086S；RG7450，其靶向STEAP-1 (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；立伐土珠單抗維多汀(Lifastuzumab Vedotin)，亦稱為DNIB0600A；NaPi2b ADC；RG7599，其靶向NaPi2b (Roche-Genentech)；索菲土珠單抗維多汀(Sofituzumab vedotin)，亦稱為DMUC5754A；RG7458，其靶向MUC16 (Seagen (Seattle Genetics) Inc.；Roche-Genentech)；RG7841，亦稱為DLYE5953A，其靶向Ly6E (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；RG7598，亦稱為DFRF4539A，其靶向FCRH5 (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；RG7636，亦稱為DEDN6526A，其靶向ETBR (Seagen (Seattle Genetics) Inc.；Roche-Genentech)；匹納土珠單抗維多汀(Pinatuzumab vedotin)，亦稱為DCDT2980S；RG7593，其靶向CD22 (Roche-Genentech)；波妥珠單抗維多汀(Polatuzumab vedotin)，亦稱為DCDS4501A；POLIVY™；RG7596；RO-5541077，其靶向CD79b (Chugai Pharmaceutical；Roche-Genentech；Seagen (Seattle Genetics) Inc.)；DMUC4064A，亦稱為D-4064a；RG7882，其靶向MUC16 (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；SYSA1801，亦稱為CPO102，其靶向CLDN18.2 (Conjupro Biotherapeutics Inc.；CSPC ZhongQi Pharmaceutical Technology Co.)；RC118，亦稱為密連蛋白18.2-ADC；YH005，其靶向CLDN18.2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)；Biocytogen)；VLS-101，亦稱為瑟吐珠單抗維多汀(Cirmtuzumab vedotin)；MK-2140；UC-961ADC3；澤羅妥單抗維多汀(Zilovertamab Vedotin)，其靶向ROR1 (VelosBio. Inc)；格雷巴單抗維多汀(Glembatumumab vedotin)，亦稱為CDX-011；CR011-vcMMAE，其靶向gpNMB (Celldex Therapeutics)；BA3021，亦稱為CAB-ROR2-ADC；奧珠單抗維多汀(Ozuriftamab Vedotin)，其靶向ROR2 (Bioatla；Himalaya Therapeutics)；BA3011，亦稱為CAB-AXL-ADC；美克波單抗維多汀(Mecbotamab Vedotin)，其靶向Axl (Bioatla; Himalaya Therapeutics)；CM-09，亦稱為Bstrongximab-ADC，其靶向TRA-1-60 (CureMeta)；ABBV-838，亦稱為阿妥昔珠單抗維多汀(Azintuxizumab vedotin)，其靶向SLAMF7 (Abbvie)；依那波他單抗維多汀(Enapotamab vedotin)，亦稱為AXL-107-MMAE；HuMax-AXL-ADC，其靶向Axl (GenMab; Seagen (Seattle Genetics) Inc.)；ARC-01，亦稱為抗CD79b ADC，其靶向CD79b (Araris Biotech AG)；迪西妥單抗維多汀(Disitamab vedotin)，亦稱為Aidexi®；RC48，其靶向HER-2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)；Seagen (Seattle Genetics) Inc.)；ASG-5ME，亦稱為AGS-5；AGS-5ME，其靶向SLC44A4 (Agensys, Inc.；Astellas Pharma Inc.；Seagen (Seattle Genetics) Inc.)；恩諾單抗維多汀(Enfortumab vedotin)，亦稱為AGS-22M6E；ASG-22CE；ASG-22ME；PADCEV™，其靶向連接素-4 (Astellas Pharma Inc.；Seagen (Seattle Genetics) Inc.)；ASG-15ME，亦稱為AGS-15E；斯妥尤單抗維多汀(Sirtratumab vedotin)，其靶向SLITRK6 (Seagen (Seattle Genetics) Inc.；Astellas Pharma Inc.)；本妥昔單抗維多汀(Brentuximab vedotin)，亦稱為Adcetris；cAC10-vcMMAE；SGN-35，其靶向CD30 (Seagen (Seattle Genetics) Inc.；Takeda)；肽利索單抗維多汀(Telisotuzumab vedotin)，亦稱為ABBV-399，其靶向c-MET (Abbvie)；羅妥昔珠單抗維多汀(Losatuxizumab vedotin)，亦稱為ABBV-221，其靶向EGFR (Abbvie)；CX-2029，亦稱為ABBV-2029，其靶向CD71 (Abbvie；CytomX Therapeutics)；AB-3A4-ADC，亦稱為AB-3A4-vcMMAE，其靶向KAAG-1 (Alethia Biotherapeutics)；因杜薩土單抗維多汀(Indusatumab vedotin)，亦稱為5F9-vcMMAE；MLN0264；TAK-264，其靶向GCC (Takeda；Millennium Pharmaceuticals, Inc)；FOR46，其靶向CD46 (Fortis Therapeutics, Inc.)；LR004-VC-MMAE，其靶向EGFR (Chinese Academy of Medical Sciences Peking Union Medical College Hospital)；CD30-ADC，其靶向CD30 (NBE Therapeutics；Boehringer Ingelheim)；抗內皮唾酸蛋白-MC-VC-PABC-MMAE，其靶向CD248 (Genzyme)；OBI-998，其靶向SSEA-4 (OBI Pharma)；MRG002，其靶向HER-2 (Lepu biotech；Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd))；TRS005，其靶向CD20 (Teruisi Pharmaceuticals)；Oba01，其靶向DR5 (死亡受體5) (Obio Technology (Shanghai) Corp.,Ltd.；Yantai Obioadc Biomedical Technology Ltd.)；PSMA ADC，其靶向PSMA (Progenics Pharmaceuticals, Inc；Seagen (Seattle Genetics) Inc.)；SGN-CD48A，其靶向CD48 (Seagen (Seattle Genetics) Inc.)；IMAB362-vcMMAE，其靶向CLDN18.2 (Astellas Pharma Inc.; Ganymed)；GB251，其靶向HER-2 (Genor Biopharma Co., Ltd.)；Innate Pharma BTG-ADC，其靶向CD30 (Innate Pharma; Sanofi)；ADCendo uPARAP ADC，其靶向MRC2 (ADCendo)；XCN-010，其靶向actM (Xiconic Pharmaceuticals, LLC)；ANT-043，其靶向HER-2 (Antikor Biopharma)；OBI-999，其靶向Globo H (Abzena; OBI Pharma)；LY3343544，其靶向MET (Eli Lilly and Company)；Tagworks抗TAG72 ADC，其靶向TAG-72 (Tagworks Pharmaceuticals)；IMAB027-vcMMAE，其靶向CLDN6 (Ganymed; Astellas Pharma Inc.)；LGR5-ADC，其靶向LGR5 (Genentech, Inc.)；Philochem B12-MMAE ADC，其靶向IL-7R (Instituto de Medicina Molecular João Lobo Antunes; Philochem AG)；TE-1522，其靶向CD19 (Immunwork)；SGN-STNV，其靶向STn (Seagen (Seattle Genetics) Inc.)；HTI-1511，其靶向EGFR (Abzena; Halozyme Therapeutics)；Peptron PAb001-ADC，其靶向OT-MUC1 (腫瘤栓繫-MUC1) (Peptron; Qilu Pharmaceutical co. Ltd.)；LM-102，其靶向CLDN18.2 (LaNova Medicines Limited)；Anwita Biosciences MSLN-MMAE，其靶向間皮素(MSLN) (Anwita biosciences)；SGN-CD228A，其靶向CD228 (Seagen (Seattle Genetics) Inc.)；NBT828，其靶向HER-2 (NewBio Therapeutics; Genor Biopharma Co., Ltd.)；Gamamabs GM103，其靶向AMHR2 (GamaMabs Pharma; Exelixis)；LCB14-0302，其靶向HER-2 (Lego Chem Biosciences)；BAY79-4620，其靶向碳酸酐酶IX (CAIX) (Bayer; MorphoSys)；NBT508，其靶向CD79b (NewBio Therapeutics)；PAT-DX3-MMAE，其靶向未揭示(Patrys; Yale University)；AGS67E，其靶向CD37 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.)；CDX-014，其靶向TIM-1 (Celldex Therapeutics)；BVX001，其靶向CD33；CD7 (Bivictrix therapeutics)；SGN-B6A，其靶向整合素β-6 (Seagen (Seattle Genetics) Inc.)；MRG003，其靶向EGFR (Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd))及PYX-202，其靶向DLK-1 (Pyxis Oncology; Lego Chem Biosciences)。Embodiment 33-1. The method of embodiment 33, wherein the antibody-drug conjugate comprises MMAE and is selected from: DP303c, also known as SYSA1501, which targets HER-2 (CSPC Pharmaceutical; Dophen Biomed); SIA01-ADC , also known as ST1, which targets STn (Siamab Therapeutics); Ladiratuzumab vedotin, also known as SGN-LIV1A, which targets LIV-1 (Merck & Co., Inc. ; Seagen (Seattle Genetics) Inc.); ABBV-085, also known as Samrotamab vedotin, which targets LRRC15 (Abbvie; Seagen (Seattle Genetics) Inc.); DMOT4039A, also known as is RG7600; αMSLN-MMAE, which targets mesothelin (MSLN) (Roche-Genentech); RC68, also known as Remegen EGFR ADC, which targets EGFR (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)) RC108, also known as RC108-ADC, which targets c-MET (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)); CMG901, also known as MRG005, which targets CLDN18.2 (Keymed Biosciences; Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd)); YBL-001, also known as LCB67, which targets DLK-1 (Lego Chem Biosciences; Pyxis Oncology; Y-Biologics); DCDS0780A, also known as Iladatuzumab vedotin; RG7986, which targets CD79b (Roche-Genentech; Seagen (Seattle Genetics) Inc.); Tisotumab vedotin, also known as Humax - TF-ADC; tf-011-mmae; TIVDAK™, which targets CD142 (GenMab; Seagen (Seattle G enetics) Inc.); GO-3D1-ADC, also known as humAb-3D1-MMAE ADC, which targets MUCl-C (Genus Oncology LLC); ALT-P7, also known as HM2-MMAE, which targets HER- 2 (Alteogen, Inc.; Levena Biopharma; 3SBio, Inc.); Vandortuzumab vedotin, also known as DSTP3086S; RG7450, which targets STEAP-1 (Roche-Genentech; Seagen ( Seattle Genetics) Inc.); Lifastuzumab Vedotin, also known as DNIB0600A; NaPi2b ADC; RG7599, which targets NaPi2b (Roche-Genentech); (Sofituzumab vedotin), also known as DMUC5754A; RG7458, which targets MUC16 (Seagen (Seattle Genetics) Inc.; Roche-Genentech); RG7841, also known as DLYE5953A, which targets Ly6E (Roche-Genentech; Seagen (Seattle Genetics) ) Inc.); RG7598, also known as DFRF4539A, which targets FCRH5 (Roche-Genentech; Seagen (Seattle Genetics) Inc.); RG7636, also known as DEDN6526A, which targets ETBR (Seagen (Seattle Genetics) Inc.); Roche-Genentech); Pinatuzumab vedotin, also known as DCDT2980S; RG7593, which targets CD22 (Roche-Genentech); Polatuzumab vedotin, also known as Known as DCDS4501A; POLIVY™; RG7596; RO-5541077, which targets CD79b (Chugai Pharmaceutical; Roche-Genentech; Seagen (Seattle Genetics) Inc.); DMUC4064A, also known as D-4064a; RG7882, which targets MUC16 ( Roche-Genentech; Seagen (Seattle Geneti cs) Inc.); SYSA1801, also known as CPO102, which targets CLDN18.2 (Conjupro Biotherapeutics Inc.; CSPC ZhongQi Pharmaceutical Technology Co.); RC118, also known as claudin 18.2-ADC; YH005, which targets CLDN18.2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.); Biocytogen); VLS-101, also known as Cirmtuzumab vedotin; MK-2140; UC-961ADC3; Zilvertamab Vedotin, which targets ROR1 (VelosBio. Inc); Glembatumumab vedotin, also known as CDX-011; CR011-vcMMAE, which targets gpNMB ( Celldex Therapeutics); BA3021, also known as CAB-ROR2-ADC; Ozuriftamab Vedotin, which targets ROR2 (Bioatla; Himalaya Therapeutics); BA3011, also known as CAB-AXL-ADC; Mecbotamab Vedotin, which targets Axl (Bioatla; Himalaya Therapeutics); CM-09, also known as Bstrongximab-ADC, which targets TRA-1-60 (CureMeta); ABBV-838, Also known as Azintuxizumab vedotin, which targets SLAMF7 (Abbvie); Enapotamab vedotin, also known as AXL-107-MMAE; HuMax-AXL - ADC, which targets Axl (GenMab; Seagen (Seattle Genetics) Inc.); ARC-01, also known as anti-CD79b ADC, which targets CD79b (Araris Biotech AG); disitumab vedotin (Disitamab) vedotin), also known as Aidexi®; RC48, which targets HER-2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.); Seagen (Seattle Genetics) Inc.); ASG-5ME, also known as AGS-5; AGS-5ME, which targets SLC44A4 (Agensys, Inc.; Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.); Enfortumab vedotin, also known as AGS-22M6E; ASG-22CE; ASG-22ME; PADCEV™, which targets connexin-4 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.); ASG -15ME, also known as AGS-15E; Sirtratumab vedotin, which targets SLITRK6 (Seagen (Seattle Genetics) Inc.; Astellas Pharma Inc.); Bentuximab vedotin (Brentuximab vedotin), also known as Adcetris; cAC10-vcMMAE; SGN-35, which targets CD30 (Seagen (Seattle Genetics) Inc.; Takeda); Telisotuzumab vedotin, also known as ABBV -399, which targets c-MET (Abbvie); Losatuxizumab vedotin, also known as ABBV-221, which targets EGFR (Abbvie); CX-2029, also known as ABBV -2029, which targets CD71 (Abbvie; CytomX Therapeutics); AB-3A4-ADC, also known as AB-3A4-vcMMAE, which targets KAAG-1 (Alethia Biotherapeutics); Indusatumumab vedotin ( Indusatumab vedotin), also known as 5F9-vcMMAE; MLN0264; TAK-264, which targets GCC (Takeda; Millennium Pharmaceuticals, Inc); FOR46, which targets CD46 (Fortis Therapeutics, Inc.); LR004-VC-MMAE, It targets EGFR (Chinese Academy of Medical Sciences Peking Union Medical College Hospital); CD30-ADC, which targets CD30 (NB E Therapeutics; Boehringer Ingelheim); anti-endosialin-MC-VC-PABC-MMAE, which targets CD248 (Genzyme); OBI-998, which targets SSEA-4 (OBI Pharma); MRG002, which targets HER -2 (Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd)); TRS005, which targets CD20 (Teruisi Pharmaceuticals); Oba01, which targets DR5 (death receptor 5) (Obio Technology (Shanghai) Corp., Ltd.; Yantai Obioadc Biomedical Technology Ltd.); PSMA ADC, which targets PSMA (Progenics Pharmaceuticals, Inc; Seagen (Seattle Genetics) Inc.); SGN-CD48A, which targets CD48 (Seagen (Seattle Genetics)) Inc.); IMAB362-vcMMAE, which targets CLDN18.2 (Astellas Pharma Inc.; Ganymed); GB251, which targets HER-2 (Genor Biopharma Co., Ltd.); Innate Pharma BTG-ADC, which targets CD30 (Innate Pharma; Sanofi); ADCendo uPARAP ADC, which targets MRC2 (ADCendo); XCN-010, which targets actM (Xiconic Pharmaceuticals, LLC); ANT-043, which targets HER-2 (Antikor Biopharma); OBI-999, which targets Globo H (Abzena; OBI Pharma); LY3343544, which targets MET (Eli Lilly and Company); Tagworks anti-TAG72 ADC, which targets TAG-72 (Tagworks Pharmaceuticals); IMAB027-vcMMAE, which Targets CLDN6 (Ganymed; Astellas Pharma Inc.); LGR5-ADC, which targets LGR5 (Genentech, Inc.); Philochem B12 - MMAE ADC, which targets IL-7R (Instituto de Medicina Molecular João Lobo Antunes; Philochem AG); TE-1522, which targets CD19 (Immunwork); SGN-STNV, which targets STn (Seagen (Seattle Genetics) Inc) .); HTI-1511, which targets EGFR (Abzena; Halozyme Therapeutics); Peptron PAb001-ADC, which targets OT-MUC1 (Tumor Tethered-MUC1) (Peptron; Qilu Pharmaceutical co. Ltd.); LM-102 , which targets CLDN18.2 (LaNova Medicines Limited); Anwita Biosciences MSLN-MMAE, which targets mesothelin (MSLN) (Anwita biosciences); SGN-CD228A, which targets CD228 (Seagen (Seattle Genetics) Inc.) NBT828, which targets HER-2 (NewBio Therapeutics; Genor Biopharma Co., Ltd.); Gamamabs GM103, which targets AMHR2 (GamaMabs Pharma; Exelixis); LCB14-0302, which targets HER-2 (Lego Chem Biosciences ); BAY79-4620, which targets carbonic anhydrase IX (CAIX) (Bayer; MorphoSys); NBT508, which targets CD79b (NewBio Therapeutics); PAT-DX3-MMAE, which targets undisclosed (Patrys; Yale University) AGS67E, which targets CD37 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.); CDX-014, which targets TIM-1 (Celldex Therapeutics); BVX001, which targets CD33; CD7 (Bivictrix therapeutics); SGN-B6A, which targets integrin beta-6 (Seagen (Seattle Genetics) Inc.); MRG003, which targets EGFR (Lepu biotech; Shanghai Miracogen) Inc. (Shanghai Meiya Biotechnology Co., Ltd)) and PYX-202, which targets DLK-1 (Pyxis Oncology; Lego Chem Biosciences).

實施例34.如實施例33之方法，其中MMAE經由包含纈胺酸及瓜胺酸之連接子與第一抗體結合。Embodiment 34. The method of embodiment 33, wherein the MMAE is conjugated to the first antibody via a linker comprising valine and citrulline.

實施例35.如實施例34之方法，其中連接子-MMAE為vcMMAE。Embodiment 35. The method of Embodiment 34, wherein the linker-MMAE is vcMMAE.

實施例36.如實施例33之方法，其中MMAE經由包含白胺酸、丙胺酸及麩胺酸之連接子與第一抗體結合。Embodiment 36. The method of embodiment 33, wherein the MMAE is conjugated to the first antibody via a linker comprising leucine, alanine, and glutamic acid.

實施例37.如實施例36之方法，其中連接子-MMAE為dLAE-MMAE。Embodiment 37. The method of Embodiment 36, wherein the linker-MMAE is dLAE-MMAE.

實施例38.如實施例1至32中任一項之方法，其中微管蛋白干擾劑為MMAF。Embodiment 38. The method of any one of embodiments 1-32, wherein the tubulin interfering agent is MMAF.

實施例38-1.如實施例38之方法，其中抗體-藥物結合物包含MMAF且係選自：CD70-ADC，其靶向CD70 (Kochi University; Osaka University)；IGN786，其靶向SAIL (AstraZeneca；Igenica Biotherapeutics)；PF-06263507，其靶向5T4 (Pfizer)；GPC1-ADC，其靶向GPC-1 (Kochi University)；ADC-AVP10，其靶向CD30 (Avipep)；M290-MC-MMAF，其靶向CD103 (The Second Affiliated Hospital of Harbin Medical University)；BVX001，其靶向CD33；CD7 (Bivictrix therapeutics)；Tanabe P3D12-vc-MMAF，其靶向c-MET (Tanabe Research Laboratories)；LILRB4-靶向ADC，其靶向LILRB4 (The University of Texas Health Science Center, Houston)；TSD101，亦稱為ABL201，其靶向BCMA (TSD Life Science；ABL Bio；Lego Chem Biosciences)；瑪汀-迪妥昔珠單抗莫福汀(Depatuxizumab mafodotin)，亦稱為ABT-414，其靶向EGFR (Abbvie; Seagen (Seattle Genetics) Inc.)；AGS16F，亦稱為AGS-16C3F；AGS-16M8F，其靶向ENPP3 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.)；AVG-A11 BCMA ADC，亦稱為AVG-A11-mcMMAF，其靶向BCMA (Avantgen)；貝蘭他單抗莫福汀(Belantamab mafodotin)，亦稱為BLENREP；GSK2857916；J6M0-mcMMAF，其靶向BCMA (GlaxoSmithKline；Seagen (Seattle Genetics) Inc.)；MP-HER3-ADC，亦稱為HER3-ADC，其靶向HER-3 (MediaPharma)；FS-1502，亦稱為LCB14-0110，其靶向HER-2 (Lego Chem Biosciences;Shanghai Fosun Pharmaceutical Development Co, Ltd.)；MEDI-547，亦稱為MI-CP177，其靶向EphA2 (AstraZeneca；Seagen (Seattle Genetics) Inc.)；伏司妥珠單抗莫福汀(Vorsetuzumab mafodotin)，亦稱為SGN-75，其靶向CD70 (Seagen (Seattle Genetics) Inc.)；德甯土珠單抗莫福汀(Denintuzumab mafodotin)，亦稱為SGN-CD19A，其靶向CD19 (Seagen (Seattle Genetics) Inc.)及HTI-1066，亦稱為SHR-A1403，其靶向c-MET (Jiangsu HengRui Medicine Co., Ltd)。Embodiment 38-1. The method of embodiment 38, wherein the antibody-drug conjugate comprises MMAF and is selected from: CD70-ADC, which targets CD70 (Kochi University; Osaka University); IGN786, which targets SAIL (AstraZeneca Igenica Biotherapeutics); PF-06263507, which targets 5T4 (Pfizer); GPC1-ADC, which targets GPC-1 (Kochi University); ADC-AVP10, which targets CD30 (Avipep); M290-MC-MMAF, It targets CD103 (The Second Affiliated Hospital of Harbin Medical University); BVX001, which targets CD33; CD7 (Bivictrix therapeutics); Tanabe P3D12-vc-MMAF, which targets c-MET (Tanabe Research Laboratories); LILRB4-targeting To ADC, which targets LILRB4 (The University of Texas Health Science Center, Houston); TSD101, also known as ABL201, which targets BCMA (TSD Life Science; ABL Bio; Lego Chem Biosciences); Martin-Dituximab Depatuxizumab mafodotin, also known as ABT-414, which targets EGFR (Abbvie; Seagen (Seattle Genetics) Inc.); AGS16F, also known as AGS-16C3F; AGS-16M8F, which targets ENPP3 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.); AVG-A11 BCMA ADC, also known as AVG-A11-mcMMAF, which targets BCMA (Avantgen); Belantamab mafodotin , also known as BLENREP; GSK2857916; J6M0-mcMMAF, which targets BCMA (GlaxoSmithKline; Seagen (Seattle Genetics) Inc.); MP-HER3-ADC, also known as HER3-ADC, which targets HER-3 (MediaPharma) ;FS-1 502, also known as LCB14-0110, which targets HER-2 (Lego Chem Biosciences; Shanghai Fosun Pharmaceutical Development Co, Ltd.); MEDI-547, also known as MI-CP177, which targets EphA2 (AstraZeneca; Seagen ( Seattle Genetics) Inc.); Vorsetuzumab mafodotin, also known as SGN-75, which targets CD70 (Seagen (Seattle Genetics) Inc.); Denin (Seattle Genetics) Inc.); Denintuzumab mafodotin), also known as SGN-CD19A, which targets CD19 (Seagen (Seattle Genetics) Inc.) and HTI-1066, also known as SHR-A1403, which targets c-MET (Jiangsu HengRui Medicine Co., Ltd ).

實施例39.如實施例1至30中任一項之方法，其中微管蛋白干擾劑為微管蛋白裂解素。Embodiment 39. The method of any one of embodiments 1-30, wherein the tubulin interfering agent is tubulin lysin.

實施例40.如實施例39之方法，其中微管蛋白裂解素係選自微管蛋白裂解素D、微管蛋白裂解素M、微管苯丙胺酸及微管酪胺酸。Embodiment 40. The method of embodiment 39, wherein the tubulin lysin is selected from the group consisting of tubulin D, tubulin M, tubulin phenylalanine, and tubulin tyrosine.

實施例41.如實施例1至32中任一項之方法，其中抗體-藥物結合物選自AbGn-107 (Ab1-18Hr1)、AGS62P1 (ASP1235)、ALT-P7 (HM2-MMAE)、BA3011 (CAB-AXL-ADC)、貝蘭單抗莫福汀、本妥昔單抗維多汀、瑟吐珠單抗維多汀(VLS-101、UC-961ADC3)、考非妥珠單抗比利多汀(cofetuzumab pelidotin) (PF-06647020、PTK7-ADC、PF-7020、ABBV-647)、CX-2029 (ABBV-2029)、迪西妥單抗維多汀(RC48)、伊那波它單抗維多汀(HuMax-AXL-ADC、AXL-107-MMAE)、恩諾單抗維多汀(EV)、FS-1502 (LCB14-0110)、吉妥單抗奧佐米星(gemtuzumab ozogamicin)、HTI-1066 (SHR-A1403)、英妥珠單抗奧佐米星(inotuzumab ozogamicin)、PF-06804103 (NG-HER2 ADC)、波妥珠單抗維多汀、薩西土珠單抗戈維特坎(sacituzumab govitecan)、SGN-B6A、SGN-CD228A、SGN-STNV、STI-6129 (CD38 ADC、LNDS1001、CD38-077 ADC)、肽利索單抗維多汀(ABBV-399)、替索圖單抗維多汀(Humax-TF-ADC、tf-011-mmae、TV)、曲妥珠單抗德魯特坎(trastuzumab deruxtecan)、曲妥珠單抗恩他新(trastuzumab emtansine)及伏司妥珠單抗莫福汀。Embodiment 41. The method of any one of embodiments 1 to 32, wherein the antibody-drug conjugate is selected from the group consisting of AbGn-107 (Ab1-18Hr1), AGS62P1 (ASP1235), ALT-P7 (HM2-MMAE), BA3011 ( CAB-AXL-ADC), belantuzumab mofotine, brentuximab vedotin, cetozumab vedotin (VLS-101, UC-961ADC3), cofetuzumab bilidol cofetuzumab pelidotin (PF-06647020, PTK7-ADC, PF-7020, ABBV-647), CX-2029 (ABBV-2029), dicituzumab pelidotin (RC48), enabotazumab Dotin (HuMax-AXL-ADC, AXL-107-MMAE), ennozumab vedotin (EV), FS-1502 (LCB14-0110), gemtuzumab ozogamicin (gemtuzumab ozogamicin), HTI -1066 (SHR-A1403), inotuzumab ozogamicin (inotuzumab ozogamicin), PF-06804103 (NG-HER2 ADC), pertuzumab vedotin, sacituzumab govitcan ( sacituzumab govitecan), SGN-B6A, SGN-CD228A, SGN-STNV, STI-6129 (CD38 ADC, LNDS1001, CD38-077 ADC), peptidolizumab vedotin (ABBV-399), tesotuzumab vedotin Dotin (Humax-TF-ADC, tf-011-mmae, TV), trastuzumab deruxtecan, trastuzumab emtansine, and vostuzumab Anti-mofatine.

實施例42.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗密連蛋白-18.2抗體，其包含分別包含SEQ ID NO:61至66之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 42. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-claudin-18.2 antibody comprising an antibody comprising SEQ ID NOs: 61 to 66, respectively Heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 of amino acid sequences.

實施例43.如實施例42之方法，其中抗密連蛋白-18.2抗體包含：包含SEQ ID NO: 59之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO: 60之胺基酸序列的輕鏈可變區(VL)。Embodiment 43. The method of embodiment 42, wherein the anti-claudin-18.2 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:59; and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:60 The light chain variable region (VL) of the amino acid sequence.

實施例44.如實施例43之方法，其中抗密連蛋白-18.2抗體為唑貝妥西單抗(zolbetuximab) (175D10)。Embodiment 44. The method of embodiment 43, wherein the anti-claudin-18.2 antibody is zolbetuximab (175D10).

實施例45.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗密連蛋白-18.2抗體，其包含分別包含SEQ ID NO:69至74之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 45. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-claudin-18.2 antibody comprising an antibody comprising SEQ ID NOs: 69 to 74, respectively Heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 of amino acid sequences.

實施例46.如實施例45之方法，其中抗密連蛋白-18.2抗體包含：包含SEQ ID NO:67之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:68之胺基酸序列的輕鏈可變區(VL)。Embodiment 46. The method of embodiment 45, wherein the anti-Clamectin-18.2 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:67; and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:68 The light chain variable region (VL) of the amino acid sequence.

實施例47.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗PD-L1抗體，其包含分別包含SEQ ID NO:77至82之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 47. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-PD-L1 antibody comprising an amine group comprising SEQ ID NOs: 77 to 82, respectively Heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 of acid sequence.

實施例48.如實施例47之方法，其中抗PD-L1抗體包含：包含SEQ ID NO:75之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:76之胺基酸序列的輕鏈可變區(VL)。Embodiment 48. The method of embodiment 47, wherein the anti-PD-L1 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:75; and an amino group comprising SEQ ID NO:76 The light chain variable region (VL) of the acid sequence.

實施例49.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗ALP抗體，其包含分別包含SEQ ID NO:85至90之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 49. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-ALP antibody comprising amino acid sequences comprising SEQ ID NOs: 85 to 90, respectively The heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3.

實施例50.如實施例49之方法，其中抗ALP抗體包含：包含SEQ ID NO:83之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:84之胺基酸序列的輕鏈可變區(VL)。Embodiment 50. The method of embodiment 49, wherein the anti-ALP antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:83; and the amino acid sequence comprising SEQ ID NO:84 The light chain variable region (VL).

實施例51.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體包含抗B7H4抗體，其包含分別包含SEQ ID NO:93至98之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 51. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody comprises an anti-B7H4 antibody comprising amino acid sequences comprising SEQ ID NOs: 93 to 98, respectively The heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3.

實施例52.如實施例51之方法，其中抗B7H4抗體包含：包含SEQ ID NO:91之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:92之胺基酸序列的輕鏈可變區(VL)。Embodiment 52. The method of embodiment 51, wherein the anti-B7H4 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:91; and the amino acid sequence comprising SEQ ID NO:92 The light chain variable region (VL).

實施例53.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗HER2抗體，其包含：包含SEQ ID NO:99之胺基酸序列的重鏈及包含SEQ ID NO:100之胺基酸序列的輕鏈。Embodiment 53. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-HER2 antibody, comprising: a heavyweight comprising the amino acid sequence of SEQ ID NO:99 chain and light chain comprising the amino acid sequence of SEQ ID NO:100.

實施例54.如實施例53之方法，其中抗體-藥物結合物為迪西妥單抗維多汀。Embodiment 54. The method of Embodiment 53, wherein the antibody-drug conjugate is dicituzumab vedotin.

實施例55.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗NaPi2B抗體，其包含：包含SEQ ID NO:101之胺基酸序列的重鏈及包含SEQ ID NO:102之胺基酸序列的輕鏈。Embodiment 55. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-NaPi2B antibody, comprising: a heavyweight comprising the amino acid sequence of SEQ ID NO: 101 chain and a light chain comprising the amino acid sequence of SEQ ID NO:102.

實施例56.如實施例55之方法，其中抗體-藥物結合物為立伐土珠單抗維多汀。Embodiment 56. The method of embodiment 55, wherein the antibody-drug conjugate is rivatuzumab vedotin.

實施例57.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗連接素-4抗體，其包含分別包含SEQ ID NO:105至110之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 57. The method of any one of embodiments 1-41, 33-1 and 38-1, wherein the first antibody is an anti-connexin-4 antibody comprising an amine comprising SEQ ID NOs: 105-110, respectively Heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 of the amino acid sequence.

實施例58.如實施例57之方法，其中抗連接素-4抗體為包含以下之抗體：包含SEQ ID NO:103之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:104之胺基酸序列的輕鏈可變區(VL)。Embodiment 58. The method of embodiment 57, wherein the anti-connexin-4 antibody is an antibody comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 103; and comprising SEQ ID NO : The light chain variable region (VL) of the amino acid sequence of 104.

實施例59.如實施例58之方法，其中抗體-藥物結合物為恩諾單抗維多汀。Embodiment 59. The method of Embodiment 58, wherein the antibody-drug conjugate is ennozumab vedotin.

實施例60.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗AVB6抗體，其包含分別包含SEQ ID NO:113至118之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 60. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-AVB6 antibody comprising amino acid sequences comprising SEQ ID NOs: 113 to 118, respectively The heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3.

實施例61.如實施例60之方法，其中抗AVB6抗體包含：包含SEQ ID NO:111之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:112之胺基酸序列的輕鏈可變區(VL)。Embodiment 61. The method of embodiment 60, wherein the anti-AVB6 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 111; and the amino acid sequence comprising SEQ ID NO: 112 The light chain variable region (VL).

實施例62.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗AVB6抗體，其包含分別包含SEQ ID NO:121至126之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 62. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-AVB6 antibody comprising amino acid sequences comprising SEQ ID NOs: 121 to 126, respectively The heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3.

實施例63.如實施例62之方法，其中抗AVB6抗體包含：包含SEQ ID NO:119之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:120之胺基酸序列的輕鏈可變區(VL)。Embodiment 63. The method of embodiment 62, wherein the anti-AVB6 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 119; and the amino acid sequence comprising SEQ ID NO: 120 The light chain variable region (VL).

實施例64.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗CD228抗體，其包含分別包含SEQ ID NO:129至134之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 64. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-CD228 antibody comprising amino acid sequences comprising SEQ ID NOs: 129 to 134, respectively The heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3.

實施例65.如實施例64之方法，其中抗CD228抗體包含：包含SEQ ID NO:127之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:128之胺基酸序列的輕鏈可變區(VL)。Embodiment 65. The method of embodiment 64, wherein the anti-CD228 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 127; and the amino acid sequence comprising SEQ ID NO: 128 The light chain variable region (VL).

實施例66.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗LIV-1抗體，其包含分別包含SEQ ID NO:137至142之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 66. The method of any one of embodiments 1 to 41, 33-1 and 38-1, wherein the first antibody is an anti-LIV-1 antibody comprising an amine group comprising SEQ ID NOs: 137 to 142, respectively Heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 of acid sequence.

實施例67.如實施例66之方法，其中抗LIV-1抗體包含：包含SEQ ID NO:135之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:136之胺基酸序列的輕鏈可變區(VL)。Embodiment 67. The method of embodiment 66, wherein the anti-LIV-1 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 135; and an amino group comprising SEQ ID NO: 136 The light chain variable region (VL) of the acid sequence.

實施例68.如實施例1至41、33-1及38-1中任一項之方法，其中第一抗體為抗組織因子抗體，其包含分別包含SEQ ID NO:145至150之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 68. The method of any one of embodiments 1-41, 33-1 and 38-1, wherein the first antibody is an anti-tissue factor antibody comprising amino acids comprising SEQ ID NOs: 145-150, respectively Sequences of heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3.

實施例69.如實施例68之方法，其中抗組織因子抗體包含：包含SEQ ID NO:143之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:144之胺基酸序列的輕鏈可變區(VL)。Embodiment 69. The method of embodiment 68, wherein the anti-tissue factor antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:143; and an amino acid comprising SEQ ID NO:144 Sequence of the light chain variable region (VL).

實施例70.如實施例69之方法，其中抗體-藥物結合物為替索圖單抗維多汀。Embodiment 70. The method of embodiment 69, wherein the antibody-drug conjugate is tesotuzumab vedotin.

實施例71.如實施例1至70、33-1及38-1中任一項之方法，其中第二抗體結合選自以下之免疫細胞接合子：抗苗勒氏激素受體(anti-Mullerian Hormone Receptor) II (AMHR2)、B7、B7H1、B7H2、B7H3、B7H4、BAFF-R、BCMA (B細胞成熟抗原)、Bst1/CD157、C5補體、CC趨化因子受體4 (CCR4)、CD123、CD137、CD19、CD20、CD25 (IL2RA)、CD276、CD278、CD3、CD32、CD33、CD37、CD38、CD4及HIV-1 gp120結合位點、CD40、CD70、CD70 (TNF受體配位體家族中之成員)、CD80、CD86、密連蛋白18.2、c-MET、CSF1R、CTLA-4、EGFR、EGFR MET原癌基因、EPHA3、ERBB2、ERBB3、FGFR2b、FLT3、GITR、糖皮質激素誘導之TNF受體(GITR)、HER2、HER3、HLA、ICOS、IDO1、IFNAR1、IFNAR2、IGF-1R、IL-3Rα (CD123)、IL-5R、IL-5Rα、LAG-3、MET原癌基因、OX40 (CD134)、PD-1、PD-L1、PD-L2、PVRIG、EBOV糖蛋白(GP)之呼吸道融合病毒(RSV)重糖基化黏蛋白樣域、恆河猴(Rh) D、唾液酸免疫球蛋白樣凝集素8 (Siglec-8)、傳信淋巴球活化分子(SLAMF7/CS1)、T細胞受體細胞毒性T淋巴球相關抗原4 (CTLA4)、TIGIT、TIM3 (HAVCR2)、Muc1之腫瘤特異性糖抗原決定基(glycoepitope) (TA-Muc1)、VSIR (VISTA)及VTCN1。Embodiment 71. The method of any one of embodiments 1-70, 33-1, and 38-1, wherein the second antibody binds to an immune cell conjugator selected from the group consisting of: anti-Mullerian hormone receptor (anti-Mullerian hormone receptor) Hormone Receptor) II (AMHR2), B7, B7H1, B7H2, B7H3, B7H4, BAFF-R, BCMA (B cell maturation antigen), Bst1/CD157, C5 complement, CC chemokine receptor 4 (CCR4), CD123, CD137, CD19, CD20, CD25 (IL2RA), CD276, CD278, CD3, CD32, CD33, CD37, CD38, CD4 and HIV-1 gp120 binding site, CD40, CD70, CD70 (part of the TNF receptor ligand family member), CD80, CD86, claudin 18.2, c-MET, CSF1R, CTLA-4, EGFR, EGFR MET proto-oncogene, EPHA3, ERBB2, ERBB3, FGFR2b, FLT3, GITR, glucocorticoid-induced TNF receptor (GITR), HER2, HER3, HLA, ICOS, IDO1, IFNAR1, IFNAR2, IGF-1R, IL-3Rα (CD123), IL-5R, IL-5Rα, LAG-3, MET proto-oncogene, OX40 (CD134) , PD-1, PD-L1, PD-L2, PVRIG, Respiratory Fusion Virus (RSV) heavily glycosylated mucin-like domain of EBOV glycoprotein (GP), Rhesus monkey (Rh) D, Sialyl immunoglobulin Tumor specificity of agglutinin-like 8 (Siglec-8), signaling lymphocyte activation molecule (SLAMF7/CS1), T cell receptor cytotoxic T lymphocyte-associated antigen 4 (CTLA4), TIGIT, TIM3 (HAVCR2), Muc1 Glycoepitope (TA-Muc1), VSIR (VISTA) and VTCN1.

實施例72.如實施例1至71、33-1及38-1中任一項之方法，其中第二抗體結合TIGIT。Embodiment 72. The method of any one of embodiments 1-71, 33-1, and 38-1, wherein the second antibody binds to TIGIT.

實施例73.如實施例72之方法，其中第二抗體包含：(a)重鏈CDR1，其包含選自SEQ ID NO:7至9之胺基酸序列；(b)重鏈CDR2，其包含選自SEQ ID NO:10至13之胺基酸序列；(c)重鏈CDR3，其包含選自SEQ ID NO:14至16之胺基酸序列；(d)輕鏈CDR1，其包含SEQ ID NO: 17之胺基酸序列；(e)輕鏈CDR2，其包含SEQ ID NO: 18之胺基酸序列；以及(f)輕鏈CDR3，其包含SEQ ID NO: 19之胺基酸序列。Embodiment 73. The method of embodiment 72, wherein the second antibody comprises: (a) heavy chain CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 7 to 9; (b) heavy chain CDR2 comprising (c) heavy chain CDR3 comprising amino acid sequences selected from SEQ ID NOs: 14 to 16; (d) light chain CDR1 comprising SEQ ID The amino acid sequence of NO: 17; (e) the light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 18; and (f) the light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 19.

實施例74.如實施例72之方法，其中第二抗體包含：包含以下序列之重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3：(a)分別SEQ ID NO: 7、10、14、17、18及19；或(b)分別SEQ ID NO: 8、11、14、17、18及19；或(c)分別SEQ ID NO: 9、12、15、17、18及19；或(d)分別SEQ ID NO: 8、13、16、17、18及19；或(e)分別SEQ ID NO: 8、12、16、17、18及19。Embodiment 74. The method of embodiment 72, wherein the second antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the following sequences: (a) SEQ ID NOs: 7, 10, 14, respectively , 17, 18 and 19; or (b) SEQ ID NOs: 8, 11, 14, 17, 18 and 19, respectively; or (c) SEQ ID NOs: 9, 12, 15, 17, 18 and 19, respectively; or (d) SEQ ID NOs: 8, 13, 16, 17, 18 and 19, respectively; or (e) SEQ ID NOs: 8, 12, 16, 17, 18 and 19, respectively.

實施例75.如實施例72之方法，其中第二抗體包含：包含選自SEQ ID NO: 1至5之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 6之胺基酸序列的輕鏈可變區。Embodiment 75. The method of embodiment 72, wherein the second antibody comprises: a heavy chain variable region comprising an amino acid sequence selected from SEQ ID NO: 1 to 5; and an amino acid comprising SEQ ID NO: 6 Sequence of the light chain variable region.

實施例76.如實施例72之方法，其中第二抗體包含：包含選自SEQ ID NO:20至24之胺基酸序列的重鏈；及包含SEQ ID NO: 25之胺基酸序列的輕鏈。Embodiment 76. The method of embodiment 72, wherein the second antibody comprises: a heavy chain comprising the amino acid sequence selected from SEQ ID NO: 20 to 24; and a light comprising the amino acid sequence of SEQ ID NO: 25 chain.

實施例77.如實施例1至71中任一項之方法，其中第二抗體結合CD40。Embodiment 77. The method of any one of embodiments 1-71, wherein the second antibody binds to CD40.

實施例78.如實施例77之方法，其中第二抗體包含：包含以下序列之重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3：(a)分別SEQ ID NO: 30、31、32、33、34及35；或(b)分別SEQ ID NO: 30、36、32、33、34及35。Embodiment 78. The method of embodiment 77, wherein the second antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the following sequences: (a) SEQ ID NOs: 30, 31, 32, respectively , 33, 34 and 35; or (b) SEQ ID NOs: 30, 36, 32, 33, 34 and 35, respectively.

實施例79.如實施例77之方法，其中第二抗體包含：包含選自SEQ ID NO: 28之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 29之胺基酸序列的輕鏈可變區。Embodiment 79. The method of embodiment 77, wherein the second antibody comprises: a heavy chain variable region comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 28; and an amino acid sequence comprising the amino acid sequence of SEQ ID NO: 29 light chain variable region.

實施例80.如實施例77之方法，其中第二抗體包含：包含SEQ ID NO: 26之胺基酸序列的重鏈；及包含SEQ ID NO: 27之胺基酸序列的輕鏈。Embodiment 80. The method of embodiment 77, wherein the second antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:26; and a light chain comprising the amino acid sequence of SEQ ID NO:27.

實施例81.如實施例1至71、33-1及38-1中任一項之方法，其中第二抗體結合CD70。Embodiment 81. The method of any one of embodiments 1-71, 33-1 and 38-1, wherein the second antibody binds to CD70.

實施例82.如實施例81之方法，其中第二抗體包含：分別包含SEQ ID NO: 53至58之序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 82. The method of embodiment 81, wherein the second antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the sequences of SEQ ID NOs: 53 to 58, respectively.

實施例83.如實施例81之方法，其中第二抗體包含：包含SEQ ID NO: 41之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 42之胺基酸序列的輕鏈可變區。Embodiment 83. The method of embodiment 81, wherein the second antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 41; and a light chain comprising the amino acid sequence of SEQ ID NO: 42 variable region.

實施例84.如實施例1至71、33-1及38-1中任一項之方法，其中第二抗體結合BCMA。Embodiment 84. The method of any one of embodiments 1-71, 33-1 and 38-1, wherein the second antibody binds to BCMA.

實施例85.如實施例84之方法，其中第二抗體包含分別包含SEQ ID NO:47至52之序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。Embodiment 85. The method of embodiment 84, wherein the second antibody comprises heavy chain CDRl, CDR2 and CDR3 and light chain CDRl, CDR2 and CDR3 comprising the sequences of SEQ ID NOs: 47-52, respectively.

實施例86.如實施例84之方法，其中第二抗體包含：包含SEQ ID NO: 45之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 46之胺基酸序列的輕鏈可變區。Embodiment 86. The method of embodiment 84, wherein the second antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 45; and a light chain comprising the amino acid sequence of SEQ ID NO: 46 variable region.

實施例87.如實施例1至86、33-1及38-1中任一項之方法，其中第二抗體為IgG1或IgG3抗體。Embodiment 87. The method of any one of embodiments 1-86, 33-1 and 38-1, wherein the second antibody is an IgGl or IgG3 antibody.

實施例88.如實施例1至87、33-1及38-1中任一項之方法，其中第二抗體包含於抗體組合物中，其中該組合物中之至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%之抗體未經岩藻糖基化。Embodiment 88. The method of any one of embodiments 1 to 87, 33-1 and 38-1, wherein the second antibody is included in an antibody composition, wherein at least 90%, at least 91%, at least 91%, At least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the antibodies are not fucosylated.

實施例89.如實施例88之方法，其中組合物中之各抗體包含與第二抗體相同的重鏈及輕鏈胺基酸序列。Embodiment 89. The method of embodiment 88, wherein each antibody in the composition comprises the same heavy and light chain amino acid sequences as the second antibody.

實施例90.如實施例1至89、33-1及38-1中任一項之方法，其中與相同同種型之對應野生型Fc相比，第二抗體之Fc增強與一或多種活化FcγR之結合，其中活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。Embodiment 90. The method of any one of embodiments 1 to 89, 33-1 and 38-1, wherein the Fc enhancement of the second antibody is associated with one or more activating FcγRs compared to a corresponding wild-type Fc of the same isotype The binding, wherein the activated FcyR comprises one or more of FcyRIIIa, FcyRIIa and/or FcyRI.

實施例91.如實施例90之方法，其中第二抗體之Fc增強與FcγRIIIa之結合。Embodiment 91. The method of embodiment 90, wherein the Fc of the second antibody enhances binding to FcγRIIIa.

實施例92.如實施例1至91、33-1及38-1中任一項之方法，其中與相同同種型之對應野生型Fc相比，第二抗體之Fc減弱與一或多種抑制性FcγR之結合。Embodiment 92. The method of any one of embodiments 1 to 91, 33-1 and 38-1, wherein the Fc of the second antibody is attenuated and one or more inhibitory as compared to a corresponding wild-type Fc of the same isotype Binding of FcγRs.

實施例93.如實施例92之方法，其中第二抗體之Fc減弱與FcγRIIb之結合。Embodiment 93. The method of embodiment 92, wherein the Fc of the second antibody attenuates binding to FcyRIIb.

實施例94.如實施例1至93、33-1及38-1中任一項之方法，其中第二抗體之Fc增強與FcγRIIIa之結合且減弱與FcγRIIb之結合。Embodiment 94. The method of any one of embodiments 1-93, 33-1 and 38-1, wherein the Fc of the second antibody enhances binding to FcyRIIIa and reduces binding to FcyRIIb.

實施例95.如實施例1至94、33-1及38-1中任一項之方法，其中第二抗體為單株抗體。Embodiment 95. The method of any one of embodiments 1-94, 33-1 and 38-1, wherein the second antibody is a monoclonal antibody.

實施例96.如實施例1至95、33-1及38-1中任一項之方法，其中第二抗體為人類化抗體或人類抗體。Embodiment 96. The method of any one of embodiments 1-95, 33-1 and 38-1, wherein the second antibody is a humanized antibody or a human antibody.

實施例97.如實施例1至96、33-1及38-1中任一項之方法，其中癌症為膀胱癌、乳癌、子宮癌、子宮頸癌、卵巢癌、前列腺癌、睪丸癌、食道癌、胃腸癌、胃癌(gastric cancer)、胰臟癌、大腸直腸癌、大腸癌、腎癌、透明細胞腎癌瘤、頭頸癌、肺癌、肺腺癌、胃癌(stomach cancer)、生殖細胞癌、骨癌、肝癌、甲狀腺癌、皮膚癌、黑素瘤、中樞神經系統贅瘤、間皮瘤、淋巴瘤、白血病、慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤、霍奇金氏淋巴瘤(Hodgkin lymphoma)、骨髓瘤或肉瘤。Embodiment 97. The method of any one of embodiments 1 to 96, 33-1 and 38-1, wherein the cancer is bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophagus cancer cancer, gastrointestinal cancer, gastric cancer, pancreatic cancer, colorectal cancer, colorectal cancer, kidney cancer, clear cell renal carcinoma, head and neck cancer, lung cancer, lung adenocarcinoma, stomach cancer, germ cell cancer, Bone cancer, liver cancer, thyroid cancer, skin cancer, melanoma, central nervous system neoplasm, mesothelioma, lymphoma, leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, Hormone Hodgkin lymphoma, myeloma or sarcoma.

實施例98.如實施例1至97、33-1及38-1中任一項之方法，其中癌症為淋巴瘤、白血病、慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤或霍奇金氏淋巴瘤。Embodiment 98. The method of any one of embodiments 1 to 97, 33-1 and 38-1, wherein the cancer is lymphoma, leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma tumor or Hodgkin's lymphoma.

實施例99.如實施例1至98、33-1及38-1中任一項之方法，其中抗體-藥物結合物及第二抗體經同時投與。Embodiment 99. The method of any one of embodiments 1-98, 33-1, and 38-1, wherein the antibody-drug conjugate and the second antibody are administered simultaneously.

實施例100.如實施例99、33-1及38-1之方法，其中抗體-藥物結合物及第二抗體以單一醫藥組合物形式投與。Embodiment 100. The method of embodiments 99, 33-1 and 38-1, wherein the antibody-drug conjugate and the second antibody are administered as a single pharmaceutical composition.

實施例101.如實施例1至98、33-1及38-1中任一項之方法，其中抗體-藥物結合物及第二抗體經依序投與。Embodiment 101. The method of any one of embodiments 1-98, 33-1, and 38-1, wherein the antibody-drug conjugate and the second antibody are administered sequentially.

實施例102.如實施例101之方法，其中至少抗體藥物結合物之第一劑量係在第二抗體之第一劑量之前投與；或其中至少第二抗體之第一劑量係在抗體藥物結合物之第一劑量之前投與。Embodiment 102. The method of embodiment 101, wherein at least the first dose of the antibody drug conjugate is administered before the first dose of the second antibody; or wherein at least the first dose of the second antibody is administered before the antibody drug conjugate administered prior to the first dose.

實施例103.如實施例1至102、33-1及38-1中任一項之方法，其中第二抗體耗盡T調節性細胞(Treg)。Embodiment 103. The method of any one of embodiments 1-102, 33-1, and 38-1, wherein the second antibody depletes T regulatory cells (Treg).

實施例104.如實施例1至103、33-1及38-1中任一項之方法，其中抗體-藥物結合物誘發針對表現由抗體藥物結合物結合之抗原的細胞的免疫記憶。Embodiment 104. The method of any one of embodiments 1-103, 33-1, and 38-1, wherein the antibody-drug conjugate induces immune memory against cells expressing the antigen bound by the antibody-drug conjugate.

實施例105.如實施例104之方法，其中免疫記憶誘導包含誘導記憶T細胞。Embodiment 105. The method of embodiment 104, wherein the induction of immune memory comprises induction of memory T cells.

實施例106.如實施例1至105、33-1及38-1中任一項之方法，其中第二抗體活化抗原呈遞細胞(APC)。Embodiment 106. The method of any one of embodiments 1-105, 33-1, and 38-1, wherein the second antibody activates an antigen presenting cell (APC).

實施例107.如實施例1至106、33-1及38-1中任一項之方法，其中第二抗體增強CD8 T細胞反應。Embodiment 107. The method of any one of embodiments 1-106, 33-1, and 38-1, wherein the second antibody enhances a CD8 T cell response.

實施例108.如實施例1至107、33-1及38-1中任一項之方法，其中第二抗體上調共刺激受體。Embodiment 108. The method of any one of embodiments 1-107, 33-1, and 38-1, wherein the second antibody upregulates a costimulatory receptor.

實施例109.如實施例1至108、33-1及38-1中任一項之方法，其中投藥ADC及第二抗體有助於釋放免疫活化細胞介素。Embodiment 109. The method of any one of embodiments 1-108, 33-1, and 38-1, wherein administering the ADC and the second antibody facilitates the release of an immune-activating interleukin.

實施例110.如實施例109之方法，其中免疫活化細胞介素為CXCL10或IFNγ。Embodiment 110. The method of embodiment 109, wherein the immune activating interleukin is CXCL10 or IFNy.

實施例111.如實施例1至110、33-1及38-1中任一項之方法，其中ADC與第二抗體協同地起作用。Embodiment 111. The method of any one of embodiments 1-110, 33-1, and 38-1, wherein the ADC acts synergistically with the second antibody.

實施例112.如實施例1至111、33-1及38-1中任一項之方法，其中組合投與ADC及第二抗體具有與以單藥療法形式投與ADC或第二抗體時相當的毒性概況。Embodiment 112. The method of any one of embodiments 1 to 111, 33-1, and 38-1, wherein administering the ADC and the second antibody in combination has the same effect as when the ADC or the second antibody is administered in monotherapy toxicity profile.

實施例113.如實施例1至112、33-1及38-1中任一項之方法，其中ADC及/或第二抗體在組合給藥時之有效劑量小於在以單藥療法形式投與時之有效劑量。Embodiment 113. The method of any one of embodiments 1-112, 33-1 and 38-1, wherein the effective dose of the ADC and/or the second antibody when administered in combination is less than when administered as monotherapy effective dose at the time.

實施例114.如實施例1至113、33-1及38-1中任一項之方法，其中癌症具有高腫瘤突變負荷。Embodiment 114. The method of any one of embodiments 1-113, 33-1, and 38-1, wherein the cancer has a high tumor mutational load.

實施例115.如實施例1至114、33-1及38-1中任一項之方法，其中癌症具有微衛星不穩定性。Embodiment 115. The method of any one of embodiments 1-114, 33-1, and 38-1, wherein the cancer has microsatellite instability.

相關申請案之交叉參考Cross-references to related applications

本申請案主張於2020年11月8日申請之美國臨時申請案第63/111,045號、2021年4月8日申請之美國臨時申請案第63/172,411號及2021年6月8日申請之美國臨時申請案第63/208,179號之優先權，其中之每一者出於任何目的以全文引用之方式併入本文中。 I.   引言 This application claims US Provisional Application No. 63/111,045, filed on November 8, 2020, US Provisional Application No. 63/172,411, filed on April 8, 2021, and US Provisional Application No. 63/172,411, filed on June 8, 2021 Priority to Provisional Application No. 63/208,179, each of which is incorporated herein by reference in its entirety for any purpose. I. Introduction

經由細胞凋亡之細胞死亡為沈默、致耐受性過程。然而，某些細胞毒素劑，包括特定抗腫瘤劑，諸如蒽環黴素、奧沙利鉑或輻射誘導細胞死亡之特徵形式，稱為免疫原性細胞死亡(ICD)。ICD為經調節細胞死亡模式/產生PBMC及T細胞針對細胞凋亡癌細胞之免疫反應。如本文中所展現，用某些微管蛋白干擾劑，諸如奧瑞他汀(例如MMAE及MMAF)進行之治療使通常發現於ER內的蛋白質暴露於細胞表面上。腫瘤抗原對T細胞之吞噬細胞吸收及呈遞增加隨後激活後天免疫系統。如本文中進一步展示，諸如MMAE及MMAF之奧瑞他汀明顯能夠驅動ICD誘導，由此允許免疫系統識別及建立針對腫瘤之細胞毒性活性。本質上，死於ICD之細胞充當刺激針對任何殘餘疾病或在復發(relapse/recurrence)情況下之腫瘤特異性免疫反應的疫苗。Cell death via apoptosis is a silent, tolerogenic process. However, certain cytotoxic agents, including certain antineoplastic agents, such as anthracyclines, oxaliplatin, or radiation induce a characteristic form of cell death known as immunogenic cell death (ICD). ICD is a modulated cell death pattern/produces PBMC and T cell immune response against apoptotic cancer cells. As demonstrated herein, treatment with certain tubulin interfering agents, such as auristatins (eg, MMAE and MMAF) exposes proteins normally found in the ER on the cell surface. Increased phagocytic uptake and presentation of tumor antigens to T cells subsequently activates the acquired immune system. As shown further herein, auristatins such as MMAE and MMAF are clearly capable of driving ICD induction, thereby allowing the immune system to recognize and establish cytotoxic activity against tumors. Essentially, cells that die from the ICD act as a vaccine to stimulate a tumor-specific immune response against any residual disease or in the case of relapse/recurrence.

如本文所描述之實驗結果所表明，回應於奧瑞他汀，諸如MMAE及MMAF之經歷ICD的腫瘤細胞顯示強化其免疫原性及細胞凋亡的獨特特徵集合，包括：鈣網伴護蛋白易位至細胞表面、在細胞凋亡期間分泌ATP及釋放核蛋白HMGB1。ICD誘導特定MAMPS及危險相關分子模式(DAMPS)之釋放，其具有建立促進腫瘤抗原之T細胞識別之促發炎環境的獨特能力。如本文所展示，儘管其他化學治療劑可誘導細胞凋亡，但並非全部均可誘導與諸如MMAE及MMAF之奧瑞他汀的ICD反應穩固性。As demonstrated by the experimental results described herein, tumor cells undergoing ICD in response to auristatins, such as MMAE and MMAF, display a unique set of features that enhance their immunogenicity and apoptosis, including: calreticulin translocation to the cell surface, secretes ATP and releases the nuclear protein HMGB1 during apoptosis. ICD induces the release of specific MAMPS and Danger-Associated Molecular Patterns (DAMPS), which have the unique ability to establish a pro-inflammatory environment that promotes T cell recognition of tumor antigens. As demonstrated herein, while other chemotherapeutic agents can induce apoptosis, not all can induce ICD response robustness to auristatins such as MMAE and MMAF.

ICD之關鍵步驟產生活化先天免疫系統以識別腫瘤細胞且清除腫瘤細胞之一系列信號。首先，藥物誘導ER應激，且其轉而引起DAMP，包括鈣網伴護蛋白、熱休克蛋白(HSP70及HSP90)之表面暴露、ATP之分泌及高遷移率基團蛋白B1 (HMGB1)之釋放。在ICD過程期間此等DAMP的暴露及免疫調節劑的分泌可共同作用以引發免疫反應，包括樹突狀細胞及其他抗原呈遞細胞之活化，導致ER應激細胞之吞噬作用及損壞。A key step in ICD generates a series of signals that activate the innate immune system to recognize and eliminate tumor cells. First, drugs induce ER stress, which in turn causes DAMPs, including calreticulin, surface exposure of heat shock proteins (HSP70 and HSP90), secretion of ATP, and release of high mobility group protein B1 (HMGB1) . Exposure of these DAMPs and secretion of immunomodulatory agents during the ICD process can act together to trigger an immune response, including activation of dendritic cells and other antigen presenting cells, leading to phagocytosis and destruction of ER stressed cells.

如上文所提及，ICD之引發與ER應激有關。過載ER去摺疊多肽或破壞蛋白質摺疊環境之能力引發ER應激反應。ER緊密連接至經由動態組裝及收縮提供結構及彈性之微管網路。微管網路之破壞對ER網路產生影響且導致嚴重ER應激，其觸發ICD誘導所需之特徵表現且引起應激反應，被稱為未摺疊蛋白反應(UPR)。UPR反應具有包括增大的pIRE1、JNK之下游磷酸化及ATF4裂解之多個臂。As mentioned above, the initiation of ICD is associated with ER stress. The ability to overload ER unfolding polypeptides or disrupt the protein folding environment triggers an ER stress response. The ER is tightly connected to a network of microtubules that provide structure and elasticity through dynamic assembly and contraction. Disruption of the microtubule network has an impact on the ER network and results in severe ER stress that triggers the characteristic manifestations required for ICD induction and induces a stress response known as the unfolded protein response (UPR). The UPR reaction has multiple arms including increased pIRE1, downstream phosphorylation of JNK, and cleavage of ATF4.

本發明係部分地基於以下發現：與其他細胞毒性劑相比且特定言之與對抗體-藥物結合物使用之其他有效負載相比，諸如奧瑞他汀(例如MMAE及MMAF)之某些微管蛋白干擾劑能夠產生獨特的ICD反應。本發明係進一步基於以下發現：將此類藥劑驅動ICD之獨特能力與增強免疫反應之藥劑配對可使抗腫瘤活性增強。尤其在使用能夠結合參與免疫傳信之某些目標且具有增強Fc結合特徵及效應功能之抗體達成此類免疫促效作用之情況下發現此發現。所需Fc結合特徵包括諸如增強與活化FcγR之結合、降低與抑制性FcγR之結合、增強ADCC活性及/或增強ADCP活性之活性。具有所需活性之某些此類抗體未經岩藻糖基化。 The present invention is based in part on the discovery that certain tubulins such as auristatins (eg MMAE and MMAF) compared to other cytotoxic agents and in particular to other payloads used for antibody-drug conjugates Interfering agents can generate unique ICD responses. The present invention is further based on the discovery that pairing the unique ability of such agents to drive ICD with an agent that enhances the immune response results in enhanced antitumor activity. This finding was particularly found in the context of such immunostimulatory effects being achieved using antibodies capable of binding certain targets involved in immune signaling and having enhanced Fc binding characteristics and effector functions. Desirable Fc binding characteristics include activities such as enhancing binding to activating FcyRs, reducing binding to inhibitory FcyRs, enhancing ADCC activity, and/or enhancing ADCP activity. Certain such antibodies with the desired activity are not fucosylated.

基於此等共同發現，如本文中更詳細地描述，本發明人已證實使用諸如MMAE及MMAF之特定微管蛋白干擾劑與亦增強Fc活性之針對免疫細胞接合子之抗體的組合誘發ICD協同地引起改良的抗腫瘤反應。使用ADC而非標準化學療法亦展示減輕傳統化學療法可見之T細胞反應的削弱，因此提供此特定組合方法之另一益處。 Based on these joint findings, as described in more detail herein, the inventors have demonstrated that induction of ICD synergistically using the combination of specific tubulin interfering agents such as MMAE and MMAF with antibodies directed against immune cell engagers that also enhance Fc activity Causes an improved antitumor response. The use of ADCs rather than standard chemotherapy has also been shown to alleviate the impairment of T cell responses seen with traditional chemotherapy, thus providing another benefit of this particular combination approach.

因此，本文提供之一些實施例為組合療法，其包含向患有癌症之個體投與：(1)抗體-藥物結合物，其包含與第一抗體結合之微管蛋白干擾劑，該第一抗體結合腫瘤相關抗原；及(2)結合於免疫細胞接合子之抗體，其中第二抗體包含增強與一或多種活化FcγR之結合的Fc。在一些實施例中，第二抗體未經岩藻糖基化。在某些實施例中，第二抗體具有增強的ADCC及/或ADCP活性。 II. 定義 Accordingly, some embodiments provided herein are combination therapies comprising administering to an individual with cancer: (1) an antibody-drug conjugate comprising a tubulin interfering agent bound to a first antibody, the first antibody an antibody that binds to a tumor-associated antigen; and (2) an antibody that binds to an immune cell engager, wherein the second antibody comprises an Fc that enhances binding to one or more activating FcγRs. In some embodiments, the second antibody is not fucosylated. In certain embodiments, the second antibody has enhanced ADCC and/or ADCP activity. II. Definitions

除非另外定義，否則本文所使用之技術及科學術語具有與一般技術者通常理解相同的含義。參見例如Lackie, DICTIONARY OF CELL AND MOLECULAR BIOLOGY, Elsevier (第4版，2007)；Sambrook 等人，MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989) 。與本文所描述之方法、裝置及材料類似或等效的任何方法、裝置及材料可用於實踐本發明。 Unless otherwise defined, technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill. See, eg, Lackie, DICTIONARY OF CELL AND MOLECULAR BIOLOGY, Elsevier (4th ed., 2007); Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989) . Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of the present invention.

如本文所用，除非上下文另外明確規定，否則單數形式「一(a/an)」及「該(the)」包括複數個參考物。因此，例如，提及「抗體」視情況包括兩個或更多個此類分子之組合及其類似物。As used herein, the singular forms "a/an" and "the" include plural references unless the context clearly dictates otherwise. Thus, for example, reference to an "antibody" optionally includes combinations of two or more such molecules and analogs thereof.

如本文所用，術語「約」係指此技術領域之技術人員易於知曉之各別值的常見誤差範圍。As used herein, the term "about" refers to the common error range for the respective value readily known to those skilled in the art.

術語「抗體」包括完整抗體及其抗原結合片段，其中抗原結合片段包含抗原結合區及位於CH2中之包含天冬醯胺(N) 297之重鏈恆定區的至少一部分。典型地，「可變區」含有抗體之抗原結合區且涉及結合之特異性及親和力。參見 Fundamental Immunology 第 7 版 ,Paul編，Wolters Kluwer Health/Lippincott Williams & Wilkins (2013)。輕鏈通常經分類為κ或λ。重鏈通常經分類為γ、μ、α、δ或ε，其轉而分別定義免疫球蛋白類別IgG、IgM、IgA、IgD及IgE。 The term "antibody" includes whole antibodies and antigen-binding fragments thereof, wherein the antigen-binding fragment comprises the antigen-binding region and at least a portion of the heavy chain constant region comprising asparagine (N) 297 located in CH2. Typically, a "variable region" contains the antigen-binding region of an antibody and is involved in specificity and affinity of binding. See Fundamental Immunology 7th Edition , Paul ed., Wolters Kluwer Health/Lippincott Williams & Wilkins (2013). Light chains are generally classified as kappa or lambda. Heavy chains are generally classified as gamma, mu, alpha, delta, or epsilon, which in turn define the immunoglobulin classes IgG, IgM, IgA, IgD, and IgE, respectively.

術語「抗體」亦包括二價或雙特異性分子、雙功能抗體、三功能抗體及四功能抗體。二價及雙特異性分子描述於例如Kostelny 等人，(1992) J. Immunol.148:1547, Pack及Pluckthun (1992) Biochemistry31:1579，Hollinger 等人(1993), PNAS. USA90:6444，Gruber 等人(1994) J Immunol. 152:5368，Zhu 等人(1997) Protein Sci. 6:781，Hu 等人(1996) Cancer Res. 56:3055，Adams 等人(1993) Cancer Res. 53:4026及McCartney, 等人(1995) Protein Eng. 8:301中。 The term "antibody" also includes bivalent or bispecific molecules, diabodies, tribodies, and tetrabodies. Bivalent and bispecific molecules are described, for example, in Kostelny et al., (1992) J. Immunol. 148:1547, Pack and Pluckthun (1992) Biochemistry 31:1579, Hollinger et al. (1993), PNAS. USA 90:6444, Gruber et al (1994) J Immunol . 152:5368, Zhu et al (1997) Protein Sci . 6:781, Hu et al (1996) Cancer Res . 56:3055, Adams et al (1993) Cancer Res . 53: 4026 and in McCartney, et al. (1995) Protein Eng . 8:301.

術語「抗體」包括抗體自身(裸抗體)或與細胞毒性或細胞生長抑制藥物結合之抗體。The term "antibody" includes the antibody itself (naked antibody) or the antibody conjugated to a cytotoxic or cytostatic drug.

「單株抗體」係指自大體上均質的抗體群獲得之抗體，亦即除可能少量存在之可能的天然存在之突變以外，構成該群體之個別抗體係相同的。修飾語「單株」指示抗體之特徵為自大體上均質之抗體群獲得，且不應理解為需要藉由任何特定方法來產生該抗體。舉例而言，欲根據本發明使用之單株抗體可藉由Kohler等人(1975) Nature 256: 495首先描述的融合瘤方法製得，或可藉由重組DNA方法(參見例如美國專利第4816567號)製得。「單株抗體」亦可使用例如Clackson等人(1991) Nature, 352:624-628及Marks等人(1991) J. Mol. Biol., 222:581-597中所描述之技術自噬菌體抗體庫分離，或可藉由其他方法製得。本文所描述之抗體為單株抗體。"Monoclonal antibody" refers to an antibody obtained from a population of substantially homogeneous antibodies, that is, identical to the individual antibody systems comprising the population, except for possible naturally occurring mutations that may be present in minor amounts. The modifier "monoclonal" indicates that the antibody is characterized as being obtained from a population of substantially homogeneous antibodies, and should not be construed as requiring the production of the antibody by any particular method. For example, monoclonal antibodies to be used in accordance with the present invention can be made by the fusionoma method first described by Kohler et al. (1975) Nature 256:495, or by recombinant DNA methods (see, eg, US Pat. No. 4,816,567). )be made of. "Monoclonal antibodies" may also be derived from phage antibody libraries using techniques such as those described in Clackson et al. (1991) Nature, 352:624-628 and Marks et al. (1991) J. Mol. Biol., 222:581-597 isolated, or can be obtained by other methods. The antibodies described herein are monoclonal antibodies.

單株抗體與其靶抗原之特異性結合意謂親和力為至少10 6、10 7、10 8、10 9或10 10M -1。特異性結合之可偵測幅度較高且可與至少一個不相關目標發生之非特異性結合區分。特異性結合可為在特定官能基或特定空間擬合(例如鎖鑰類型(lock and key type))之間形成鍵之結果，而特異性結合通常為凡得瓦爾力(van der Waals force)之結果。 Specific binding of a monoclonal antibody to its target antigen means an affinity of at least 10 6 , 10 7 , 10 8 , 10 9 or 10 10 M −1 . Specific binding has a high detectable magnitude and is distinguishable from non-specific binding occurring with at least one unrelated target. Specific binding can be the result of bond formation between specific functional groups or specific steric fit (eg, lock and key type), while specific binding is usually the result of van der Waals forces .

基本抗體結構單元為子單元之四聚體。各四聚體包括兩對相同之多肽鏈，各對具有一個「輕」鏈(約25 kDa)及一個「重」鏈(約50至70 kDa)。各鏈之胺基端部分包括主要負責抗原識別之約100至110個或更多個胺基酸之可變區。此可變區初始地表現鍵聯至可裂解信號肽。無信號肽之可變區有時被稱作成熟可變區。因此，例如輕鏈成熟可變區意謂無輕鏈信號肽之輕鏈可變區。各鏈之羧基端部分定義主要負責效應功能之恆定區。The basic antibody building block is a tetramer of subunits. Each tetramer includes two identical pairs of polypeptide chains, each pair having a "light" chain (about 25 kDa) and a "heavy" chain (about 50 to 70 kDa). The amino-terminal portion of each chain includes a variable region of about 100 to 110 or more amino acids primarily responsible for antigen recognition. This variable region initially appears to be linked to a cleavable signal peptide. Variable regions without a signal peptide are sometimes referred to as mature variable regions. Thus, for example, a light chain mature variable region means a light chain variable region without a light chain signal peptide. The carboxy-terminal portion of each chain defines the constant region primarily responsible for effector functions.

輕鏈經分類為κ或λ。重鏈經分類為γ、μ、α、δ或ε，且分別定義抗體之同型為IgG、IgM、IgA、IgD及IgE。在輕鏈及重鏈內，可變區及恆定區由具有約12或更多個胺基酸之「J」區接合，其中重鏈亦包括約10或更多個胺基酸之「D」區。(一般參見Fundamental Immunology (Paul, W.,編, 第2版，Raven Press, N.Y., 1989,第7章)，出於所有目的其以全文引用的方式併入本文中)。Light chains are classified as kappa or lambda. Heavy chains are classified as gamma, mu, alpha, delta, or epsilon, and define the antibody's isotype as IgG, IgM, IgA, IgD, and IgE, respectively. Within the light and heavy chains, the variable and constant regions are joined by a "J" region of about 12 or more amino acids, where the heavy chain also includes a "D" of about 10 or more amino acids Area. (See generally Fundamental Immunology (Paul, W., ed., 2nd ed., Raven Press, N.Y., 1989, Chapter 7), which is hereby incorporated by reference in its entirety for all purposes).

各輕鏈/重鏈對之成熟可變區形成抗體結合位點。因此，完整抗體具有兩個結合位點。不同在於在雙官能或雙特異性抗體中，兩個結合位點係相同的。各鏈均呈現藉由三個高變區接合之相對保守構架區(FR)之相同通式結構，亦稱為互補決定區或CDR。各對之兩條鏈之CDR藉由構架區對齊，使得能夠結合於特定抗原決定基。自N端至C端，輕鏈及重鏈兩者包含域FR1、CDR1、FR2、CDR2、FR3、CDR3及FR4。將胺基酸分配至各域係根據Kabat，Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987及1991)或Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987)；Chothia等人，Nature 342:878-883 (1989)之定義，或Kabat及Chothia、或IMGT (ImMunoGeneTics資訊系統)、AbM或Contact之合成或CDR之其他習知定義進行。Kabat亦提供一種廣泛使用的編號規約(Kabat編號)，其中給不同重鏈之間或不同輕鏈之間的相應殘基分配相同編號。除非上下文另外顯而易見，否則Kabat編號用於指示胺基酸在可變區中之位置。除非另外自上下文顯而易知，否則EU編號用於指示恆定區中之位置。The mature variable region of each light/heavy chain pair forms the antibody binding site. Thus, intact antibodies have two binding sites. The difference is that in bifunctional or bispecific antibodies, the two binding sites are the same. Each strand exhibits the same general structure of relatively conserved framework regions (FRs) joined by three hypervariable regions, also known as complementarity determining regions or CDRs. The CDRs of the two chains of each pair are aligned by framework regions to enable binding to specific epitopes. From the N-terminus to the C-terminus, both the light and heavy chains comprise the domains FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. Assignment of amino acids to domains is according to Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD, 1987 and 1991) or Chothia & Lesk, J. Mol. Biol. 196:901-917 (1987 ); the definition of Chothia et al., Nature 342:878-883 (1989), or the synthesis of Kabat and Chothia, or IMGT (ImMunoGeneTics Information System), AbM or Contact, or other conventional definitions of CDRs. Kabat also provides a widely used numbering convention (Kabat numbering) in which corresponding residues between different heavy chains or between different light chains are assigned the same numbering. Unless otherwise apparent from the context, Kabat numbering is used to indicate the position of the amino acid within the variable region. EU numbers are used to indicate positions in the constant region unless otherwise apparent from the context.

「人類化」抗體為保持非人類抗體之反應性同時其在人類中具較低免疫原性之抗體。此可例如藉由保留非人類CDR區且用人類對應物置換抗體之剩餘部分而獲得。 參見例如Morrison 等人， PNAS USA, 81:6851-6855 (1984)；Morrison及Oi， Adv. Immunol., 44:65-92 (1988)；Verhoeyen 等人， Science, 239:1534-1536 (1988)；Padlan, Molec. Immun., 28:489-498 (1991)；Padlan, Molec. Immun., 31(3):169-217 (1994)。 "Humanized" antibodies are antibodies that retain the reactivity of non-human antibodies while being less immunogenic in humans. This can be obtained, for example, by retaining the non-human CDR regions and replacing the remainder of the antibody with the human counterpart. See, eg , Morrison et al., PNAS USA , 81:6851-6855 (1984); Morrison and Oi, Adv. Immunol. , 44:65-92 (1988); Verhoeyen et al., Science , 239:1534-1536 (1988) ; Padlan, Molec. Immun. , 28:489-498 (1991); Padlan, Molec. Immun ., 31(3):169-217 (1994).

如本文所用，術語「嵌合抗體」係指其中(a)恆定區或其部分經置換使得抗原結合位點(可變區、CDR或其部分)鍵聯至不同物種之恆定區的抗體分子。As used herein, the term "chimeric antibody" refers to an antibody molecule in which (a) the constant region or portion thereof is substituted such that the antigen binding site (variable region, CDR or portion thereof) is linked to a constant region of a different species.

術語「抗原決定基」係指抗原上由抗體結合的位點。抗原決定基可由藉由一或多種蛋白質之三級摺疊並列的相鄰胺基酸或非相鄰胺基酸形成。由相鄰的胺基酸形成之抗原決定基通常在暴露於變性溶劑後保留，而藉由三級摺疊形成之抗原決定基通常在變性溶劑處理後消失。抗原決定基通常包括呈獨特空間構形之至少3個且更通常至少5個或8至10個胺基酸。測定抗原決定基之空間構形之方法包括例如x射線結晶學及2維核磁共振。參見例如Epitope Mapping Protocols, Methods in Molecular Biology, 第66卷, Glenn E. Morris, 編(1996)。The term "epitope" refers to the site on an antigen that is bound by an antibody. Epitopes can be formed by adjacent amino acids or non-adjacent amino acids juxtaposed by tertiary folds of one or more proteins. Epitopes formed from adjacent amino acids typically remain after exposure to denaturing solvents, whereas epitopes formed by tertiary folding typically disappear after treatment with denaturing solvents. An epitope typically includes at least 3 and more typically at least 5 or 8 to 10 amino acids in a unique spatial configuration. Methods for determining the spatial configuration of epitopes include, for example, x-ray crystallography and 2-dimensional nuclear magnetic resonance. See, eg, Epitope Mapping Protocols, Methods in Molecular Biology, Vol. 66, Glenn E. Morris, ed. (1996).

識別相同或重疊抗原決定基的抗體可以在單一免疫分析中鑑別，該單一免疫分析展示一種抗體與另一抗體競爭結合於目標抗原的能力。抗體之抗原決定基亦可以由與其抗原結合以鑑別接觸殘基之抗體的X射線結晶學定義。替代地，若抗原中減少或消除一種抗體之結合的所有胺基酸突變減少或消除了另一抗體之結合，則兩個抗體具有相同抗原決定基。若減少或消除一種抗體之結合之一些胺基酸突變減少或消除另一抗體之結合，則兩個抗體具有重疊抗原決定基。Antibodies that recognize the same or overlapping epitopes can be identified in a single immunoassay that demonstrates the ability of one antibody to compete with another antibody for binding to the target antigen. An epitope of an antibody can also be defined by X-ray crystallography of the antibody that binds to its antigen to identify contact residues. Alternatively, two antibodies have the same epitope if all amino acid mutations in the antigen that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody. Two antibodies have overlapping epitopes if some amino acid mutations that reduce or eliminate binding of one antibody reduce or eliminate binding of the other antibody.

抗體之間的競爭係藉由以下分析來測定：其中受測試抗體抑制參考抗體與共同抗原之特異性結合(參見例如Junghans等人, Cancer Res.50:1495, 1990)。若如在競爭性結合分析中所量測，過量測試抗體(例如至少2×、5×、10×、20×或100×)將參考抗體之結合抑制至少50%，但較佳75%、90%或99%，則測試抗體與參考抗體競爭。藉由競爭分析(競爭抗體)鑑別之抗體包括與參考抗體結合於相同抗原決定基之抗體及結合於足夠靠近參考抗體所結合之抗原決定基之相鄰抗原決定基以發生位阻的抗體。 Competition between antibodies is determined by assays in which the tested antibody inhibits specific binding of the reference antibody to a common antigen (see eg, Junghans et al., Cancer Res. 50:1495, 1990). An excess of test antibody (eg, at least 2×, 5×, 10×, 20× or 100×) inhibits binding of the reference antibody by at least 50%, but preferably 75%, 90%, if measured in a competitive binding assay % or 99%, the test antibody competes with the reference antibody. Antibodies identified by competition assays (competing antibodies) include antibodies that bind to the same epitope as the reference antibody and antibodies that bind to adjacent epitopes sufficiently close to the epitope to which the reference antibody binds to be sterically hindered.

片語「特異性結合」係指與分子(例如抗體或抗體片段)結合非目標化合物相比，其以對樣本中之目標更大親和力、親合力、更容易及/或以更長持續時間結合於彼目標。在一些實施例中，特異性結合目標之抗體為以比非目標化合物大至少2倍的親和力，諸如大至少4倍、5倍、6倍、7倍、8倍、9倍、10倍、20倍、25倍、50倍或100倍的親和力結合於目標之抗體。舉例而言，特異性結合TIGIT之抗體通常以比非TIGIT目標大至少2倍的親和力結合於TIGIT。一般熟習此項技術者閱讀此定義應理解，例如特異性或較佳地結合於第一目標之抗體(或部分或抗原決定基)可或可不特異性或較佳結合於第二目標。因而，「特異性結合」不必需要(但其可包括)排他性結合。The phrase "specifically binds" means that it binds with greater affinity, avidity, more readily and/or for a longer duration to a target in a sample than a molecule (eg, an antibody or antibody fragment) binds to a non-target compound on his target. In some embodiments, the antibody that specifically binds the target is at least 2-fold greater than the non-target compound, such as at least 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, 9-fold, 10-fold, 20-fold greater than the non-target compound Antibodies that bind to the target with 25-fold, 25-fold, 50-fold or 100-fold affinity. For example, antibodies that specifically bind TIGIT typically bind to TIGIT with at least 2-fold greater affinity than non-TIGIT targets. Those of ordinary skill in the art will understand this definition, for example, an antibody (or moiety or epitope) that specifically or preferably binds to a first target may or may not bind specifically or preferably to a second target. Thus, "specific binding" does not necessarily require (but may include) exclusive binding.

術語「結合親和力」在本文中用作兩個分子，例如抗體或其片段與抗原之間的非共價相互作用強度的量測。術語「結合親和力」用於描述單價相互作用(固有活性)。The term "binding affinity" is used herein as a measure of the strength of the non-covalent interaction between two molecules, eg, an antibody or fragment thereof, and an antigen. The term "binding affinity" is used to describe monovalent interactions (intrinsic activity).

兩個分子，例如抗體或其片段與抗原之間經由單價交互作用的結合親和力可藉由測定解離常數(K D)定量。繼而，作為非限制性實例，K D可藉由使用例如表面電漿子共振(SPR)方法(Biacore™)量測複合物形成及解離之動力學來測定。對應於單價複合物之締合及解離的速率常數分別稱為締合速率常數 k a (或 k on )及解離速率常數 k d (或 k off )。K D經由等式K D= k d / k a 而與 k a 及 k d 相關。解離常數之值可直接藉由熟知方法測定，且可甚至針對複合混合物藉由諸如例如Caceci等人(1984, Byte9: 340-362)中所闡述之方法來計算。舉例而言，K D可使用諸如由Wong及Lohman (1993, Proc. Natl. Acad. Sci. USA90: 5428-5432)所揭示之雙過濾器硝化纖維過濾器結合分析來確立。評估配位體，諸如抗體針對目標抗原之結合能力的其他標準分析為此項技術中已知的，包括例如ELISA、西方墨點、RIA及流式細胞量測分析及在本文中之其他地方所例示之其他分析。抗體之結合動力學及結合親和力亦可藉由此項技術中已知或如下方實例章節中所描述之標準分析評估，諸如表面電漿子共振(SPR)，例如藉由使用Biacore™系統；動力排除分析，諸如KinExA®；以及BioLayer干涉量測術(例如使用ForteBio® Octet平台)。在一些實施例中，使用BioLayer干涉量測術分析測定結合親和力。參見例如Wilson等人， Biochemistry and Molecular Biology Education, 38:400-407 (2010)；Dysinger 等人， J. Immunol. Methods, 379:30-41 (2012)；以及Estep等人， Mabs, 2013, 5:270-278。 The binding affinity between two molecules, eg, an antibody or fragment thereof, and an antigen through a monovalent interaction can be quantified by determining the dissociation constant ( KD ). Then, as a non-limiting example, KD can be determined by measuring the kinetics of complex formation and dissociation using, for example, surface plasmon resonance (SPR) methods (Biacore™). The rate constants corresponding to the association and dissociation of monovalent complexes are referred to as the association rate constant ka (or kon ) and the dissociation rate constant kd (or koff ), respectively. K D is related to ka and k d via the equation K D = k d / ka . The value of the dissociation constant can be determined directly by well-known methods, and can be calculated even for complex mixtures by methods such as those described, for example, in Caceci et al. (1984, Byte 9: 340-362). For example, KD can be established using a dual filter nitrocellulose filter binding assay such as disclosed by Wong and Lohman (1993, Proc. Natl. Acad. Sci. USA 90: 5428-5432). Other standard assays to assess the binding ability of ligands, such as antibodies, to target antigens are known in the art, including, for example, ELISA, Western blot, RIA, and flow cytometry assays and those described elsewhere herein. Examples of other analyses. Binding kinetics and binding affinity of antibodies can also be assessed by standard assays known in the art or as described in the Examples section below, such as surface plasmon resonance (SPR), for example by using the Biacore™ system; Kinetics Exclude analyses, such as KinExA®; and BioLayer interferometry (eg, using the ForteBio® Octet platform). In some embodiments, binding affinity is determined using BioLayer interferometry analysis. See, eg, Wilson et al., Biochemistry and Molecular Biology Education , 38:400-407 (2010); Dysinger et al., J. Immunol. Methods , 379:30-41 (2012); and Estep et al., Mabs , 2013, 5 : 270-278.

如本文所用，術語「交叉反應」係指抗體結合於除產生該抗體之抗原以外的抗原之能力。在一些實施例中，交叉反應性係指抗體結合於來自另一物種之抗原而非產生該抗體之抗原的能力。作為一非限制性實例，針對人類TIGIT抗原產生的如本文所描述之抗TIGIT抗體可展現與來自不同物種(例如小鼠或猴)之TIGIT的交叉反應性。As used herein, the term "cross-reactivity" refers to the ability of an antibody to bind to an antigen other than the antigen from which the antibody was raised. In some embodiments, cross-reactivity refers to the ability of an antibody to bind to an antigen from another species other than the antigen from which the antibody was raised. As a non-limiting example, anti-TIGIT antibodies as described herein raised against human TIGIT antigens can exhibit cross-reactivity with TIGIT from different species (eg, mouse or monkey).

「經分離」抗體係指已經識別且分離及/或自其天然環境之組分回收之抗體及/或以重組方式產生之抗體。「純化抗體」為通常至少50%w/w純干擾蛋白及由其產生或純化產生之其他雜質之抗體，但不排除單株抗體與過量醫藥學上可接受之載劑或其他媒劑組合以便於其使用之可能性。干擾蛋白及其他污染物可包括例如分離或以重組方式產生抗體之細胞的細胞組分。有時單株抗體為至少60%、70%、80%、90%、95%或99% w/w純之干擾蛋白及由其產生或純化產生的污染物。本文所描述之抗體，包括大鼠、嵌合、面飾化及人類化抗體可以經分離及/或純化形式提供。An "isolated" antibody system refers to an antibody and/or recombinantly produced antibody that has been recognized and isolated and/or recovered from components of its natural environment. A "purified antibody" is an antibody that is typically at least 50% w/w pure interfering protein and other impurities produced therefrom or purified, but does not exclude that the monoclonal antibody is combined with an excess of a pharmaceutically acceptable carrier or other vehicle so as to the possibility of its use. Interfering proteins and other contaminants can include, for example, cellular components of cells isolated or recombinantly producing the antibody. Sometimes a monoclonal antibody is at least 60%, 70%, 80%, 90%, 95%, or 99% w/w pure interfering protein and contaminants produced or purified therefrom. Antibodies described herein, including rat, chimeric, faceted and humanized antibodies, can be provided in isolated and/or purified form.

術語「LAE」係指三肽連接子白胺酸-丙胺酸-麩胺酸。術語「dLAE」係指三肽連接子D-白胺酸-丙胺酸-麩胺酸，其中三肽連接子中之白胺酸呈D-組態。The term "LAE" refers to the tripeptide linker leucine-alanine-glutamic acid. The term "dLAE" refers to the tripeptide linker D-leucine-alanine-glutamic acid, wherein the leucine in the tripeptide linker is in the D-configuration.

「個體(Subject)」、「患者」、「個體(individual)」及類似術語可互換使用且除指定外係指哺乳動物，諸如人類及非人類靈長類動物以及家兔、大鼠、小鼠、山羊、豬及其他哺乳動物物種。術語不一定指示個體已診斷患有特定疾病，但典型地係指處於醫療監督下之個體。"Subject," "patient," "individual," and similar terms are used interchangeably and, unless specified, refer to mammals, such as humans and non-human primates, as well as rabbits, rats, mice , goats, pigs and other mammalian species. The term does not necessarily indicate that an individual has been diagnosed with a particular disease, but typically refers to an individual under medical supervision.

術語「療法」、「治療」及「改善」係指症狀嚴重程度之任何減輕。在治療癌症的情況下，治療可指減小例如腫瘤大小、癌細胞數目、生長速率、轉移性活性、非癌細胞之細胞死亡等。如本文所用，術語「治療」及「預防」並不意欲為絕對術語。治療及預防可指任意延遲發作、減輕症狀、提高患者存活率、增加存活時間或存活比率等。治療及預防可為完全(未剩餘可偵測症狀)或部分地，使得與無本文所描述之治療的患者相比，症狀變得不太頻繁或不嚴重。可將與未接受治療之個體或個體集合與治療之前或在治療期間的不同時間點的相同患者的治療效果進行比較。在一些態樣中，如與例如投藥之前的個體或未經歷治療之個體相比，疾病嚴重程度降低至少10%。在一些態樣中，疾病嚴重程度降低至少25%、50%、75%、80%或90%，或在一些情況下，使用標準診斷技術不可再偵測到。The terms "therapy", "treatment" and "improvement" refer to any reduction in the severity of symptoms. In the context of treating cancer, treatment can refer to reducing, for example, tumor size, number of cancer cells, growth rate, metastatic activity, cell death of non-cancer cells, and the like. As used herein, the terms "treatment" and "prevention" are not intended to be absolute terms. Treatment and prevention can refer to any delay in onset, alleviation of symptoms, increase in patient survival, increase in survival time or survival rate, and the like. Treatment and prevention can be complete (no detectable symptoms remaining) or partial, such that symptoms become less frequent or less severe than in patients without the treatment described herein. The effect of treatment can be compared to an individual or set of individuals who did not receive treatment with the same patient before treatment or at different time points during treatment. In some aspects, the disease severity is reduced by at least 10% compared to, eg, a subject prior to administration or a subject not undergoing treatment. In some aspects, disease severity is reduced by at least 25%, 50%, 75%, 80%, or 90%, or in some cases, is no longer detectable using standard diagnostic techniques.

如本文所用，藥劑(例如如本文所描述之抗體)之「治療量」或「治療有效量」為藥劑預防、緩解、減輕、改善或降低個體之疾病(例如癌症)之症狀的嚴重程度的量。As used herein, a "therapeutic amount" or "therapeutically effective amount" of an agent (eg, an antibody as described herein) is an amount of the agent that prevents, alleviates, alleviates, ameliorates, or reduces the severity of symptoms of a disease (eg, cancer) in an individual .

術語「投與(administer/administered/administering)」係指將藥劑、化合物或組合物遞送至生物學作用之所需位點之方法。此等方法包括但不限於局部遞送、非經腸遞送、靜脈內遞送、真皮內遞送、肌肉內遞送、結腸遞送、直腸遞送或腹膜內遞送。可視情況與本文中所描述之藥劑及方法一起採用的投藥技術包括例如Goodman及Gilman, The Pharmacological Basis of Therapeutics,現行版, Pergamon；及Remington's, Pharmaceutical Sciences, 現行版, Mack Publishing Co., Easton, Pa中所論述。 III.      結合免疫細胞接合子之例示性抗體 The term "administer/administered/administering" refers to a method of delivering an agent, compound or composition to the desired site of biological action. Such methods include, but are not limited to, local delivery, parenteral delivery, intravenous delivery, intradermal delivery, intramuscular delivery, colonic delivery, rectal delivery, or intraperitoneal delivery. Administration techniques that may optionally be employed with the agents and methods described herein include, for example, Goodman and Gilman, The Pharmacological Basis of Therapeutics, current edition, Pergamon; and Remington's, Pharmaceutical Sciences, current edition, Mack Publishing Co., Easton, Pa. discussed in. III. Exemplary Antibodies that Bind to Immune Cell Engagers

如上文所提及，本發明人已發現藉由投與包含某些微管蛋白干擾劑(例如奧瑞他汀，包括例如MMAE及MMAF)之抗體-藥物結合物與使用抗體觸發免疫反應的組合來誘導ICD可引起改良的抗腫瘤反應，包括協同性反應，該抗體結合直接地或間接地參與免疫調節之蛋白質且具有增強的Fc活性。因而，在一些實施例中，本文所提供之方法包含向患有癌症之個體投與結合參與調節免疫反應之目標之抗體，其中此類結合誘導、促進或增強免疫反應。此類抗體之目標可稱為「免疫細胞接合子」。如本文所用之「免疫細胞接合子」係指參與積極地或不利地調節免疫細胞反應之分子(例如跨膜蛋白)。在一些實施例中，抗體結合於免疫細胞或腫瘤上之受體且引起直接免疫細胞接合或釋放陰性抑制信號。在一些實施例中，免疫細胞接合子為參與T細胞傳信之分子。在一些實施例中，免疫細胞接合子調節(例如活化)抗原呈遞細胞(APC)。在某些實施例中，免疫細胞接合子為免疫檢查點蛋白。下文列舉潛在免疫細胞接合子之其他實例。在一些實施例中，抗體結合於免疫細胞或腫瘤上之受體。結合本文所描述之免疫細胞接合子的任何抗體可與本文所描述之任何抗體-藥物結合物組合。As mentioned above, the inventors have discovered that induction of an immune response by the administration of antibody-drug conjugates comprising certain tubulin interfering agents, such as auristatin, including, for example, MMAE and MMAF in combination with the use of antibodies to trigger an immune response is induced ICD elicits improved anti-tumor responses, including synergistic responses, the antibody binds proteins directly or indirectly involved in immune regulation and has enhanced Fc activity. Thus, in some embodiments, the methods provided herein comprise administering to an individual with cancer an antibody that binds a target involved in modulating an immune response, wherein such binding induces, promotes or enhances an immune response. The targets of such antibodies may be referred to as "immune cell engagers." As used herein, an "immune cell adaptor" refers to a molecule (eg, a transmembrane protein) involved in positively or negatively modulating immune cell responses. In some embodiments, the antibody binds to a receptor on an immune cell or tumor and causes direct immune cell engagement or release of a negative inhibitory signal. In some embodiments, the immune cell zygote is a molecule involved in T cell signaling. In some embodiments, the immune cell adaptor modulates (eg activates) antigen presenting cells (APCs). In certain embodiments, the immune cell adaptor is an immune checkpoint protein. Additional examples of potential immune cell zygotes are listed below. In some embodiments, the antibody binds to a receptor on an immune cell or tumor. Any antibody that binds to the immune cell engagers described herein can be combined with any of the antibody-drug conjugates described herein.

結合參與免疫調節之目標(例如免疫細胞接合子)之抗體亦包含具有呈任何組合形式之以下特徵中之一或多者或所有的Fc：1)增強與一或多種活化FcγR之結合，2)減弱與抑制性FcγR之結合，3)未經岩藻糖基化，4)具有增強的ADCC活性，5)具有增強的ADCP活性，6)活化抗原呈遞細胞(APC)，7)增強CD8T細胞反應，8)上調共刺激受體，9)活化先天細胞免疫反應及/或10)接合NK細胞。Antibodies that bind targets involved in immune regulation (eg, immune cell engagers) also include Fcs having one or more or all of the following characteristics, in any combination: 1) enhanced binding to one or more activating FcγRs, 2) Attenuates binding to inhibitory FcγRs, 3) is not fucosylated, 4) has enhanced ADCC activity, 5) has enhanced ADCP activity, 6) activates antigen presenting cells (APCs), 7) enhances CD8 T cell responses , 8) upregulate costimulatory receptors, 9) activate innate cellular immune responses and/or 10) engage NK cells.

因此，在一些實施例中，抗體包含增強與一或多種活化FcγR之結合及/或減弱與一或多種抑制性FcγR之結合以獲得所需增強的FcγR結合概況之Fc。活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。抑制性FcγR包括例如FcγRIIb。Thus, in some embodiments, the antibody comprises an Fc that enhances binding to one or more activating FcyRs and/or reduces binding to one or more inhibitory FcyRs to achieve the desired enhanced FcyR binding profile. Activated FcyRs include one or more of FcyRIIIa, FcyRIIa, and/or FcyRI. Inhibitory FcyRs include, for example, FcyRIIb.

在某些實施例中，抗體包含增強與至少FcγRIIIa之結合的Fc。在其他實施例中，抗體包含增強與至少FcγRIIIa及FcγRIIa之結合的Fc。在一些實施例中，抗體包含增強與至少FcγRIIIa及FcγRI之結合的Fc。在某些實施例中，抗體包含增強與FcγRIIIa、FcγRIIa及FcγRI之結合的Fc。In certain embodiments, the antibody comprises an Fc that enhances binding to at least FcyRIIIa. In other embodiments, the antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRIIa. In some embodiments, the antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRI. In certain embodiments, the antibody comprises an Fc that enhances binding to FcyRIIIa, FcyRIIa, and FcyRI.

在一些實施例中，除增強與活化FcγR之結合之外或與其分離，抗體減弱與一或多種抑制性FcγR之結合。因此，在一些實施例中，抗體減弱與FcγRIIa及/或FcγRIIb之結合。In some embodiments, the antibody reduces binding to one or more inhibitory FcyRs, in addition to enhancing binding to activating FcyRs or in isolation therefrom. Thus, in some embodiments, the antibody attenuates binding to FcyRIIa and/or FcyRIIb.

在一些實施例中，抗體未經岩藻糖基化。在一些實施例中，抗體進一步具有上文所描述之FcγR結合概況中之一者。In some embodiments, the antibody is not fucosylated. In some embodiments, the antibody further has one of the FcyR binding profiles described above.

在某些實施例中，抗體之Fc包含相對於野生型Fc之胺基酸變化以增強與活化FcγR之結合及/或減弱與一或多種抑制性FcγR之結合，以獲得諸如上文所描述之FcγR結合概況。舉例而言，在一些實施例中，抗體之Fc包含重鏈恆定區中之取代S293D、A330L及I332E。In certain embodiments, the Fc of an antibody comprises amino acid changes relative to a wild-type Fc to enhance binding to activating FcγRs and/or to decrease binding to one or more inhibitory FcγRs, such as those described above FcyR binding profile. For example, in some embodiments, the Fc of the antibody comprises the substitutions S293D, A330L, and I332E in the heavy chain constant region.

下文提供關於獲得具有所需FcγR概況之抗體之方法的額外細節作為用於獲得未經岩藻糖基化抗體的方法。 A.例示性免疫細胞接合子 Additional details on methods for obtaining antibodies with the desired FcyR profiles are provided below as methods for obtaining non-fucosylated antibodies. A. Exemplary Immune Cell Conjugates

抗體可靶向之非限制性例示性目標或免疫細胞接合子包括：苗勒氏激素受體II (AMHR2)、B7、B7H1、B7H2、B7H3、B7H4、BAFF-R、BCMA (B細胞成熟抗原)、Bst1/CD157、C5補體、CC趨化因子受體4 (CCR4)、CD123、CD137、CD19、CD20、CD25 (IL2RA)、CD276、CD278、CD3、CD32、CD33、CD37、CD38、CD4及HIV-1 gp120結合位點、CD40、CD70、CD70 (TNF受體配位體家族中之成員)、CD80、CD86、密連蛋白18.2、c-MET、CSF1R、CTLA-4、EGFR、EGFR MET原癌基因、EPHA3、ERBB2、ERBB3、FGFR2b、FLT3、GITR、糖皮質激素誘導之TNF受體(GITR)、HER2、HER3、HLA、ICOS、IDO1、IFNAR1、IFNAR2、IGF-1R、IL-3Rα (CD123)、IL-5R、IL-5Rα、LAG-3、MET原癌基因、OX40 (CD134)、PD-1、PD-L1、PD-L2、PVRIG、EBOV糖蛋白(GP)之呼吸道融合病毒(RSV)重糖基化黏蛋白樣域、恆河猴(Rh) D、唾液酸免疫球蛋白樣凝集素8 (Siglec-8)、傳信淋巴球活化分子(SLAMF7/CS1)、T細胞受體細胞毒性T淋巴球相關抗原4 (CTLA4)、TIGIT、TIM3 (HAVCR2)、Muc1之腫瘤特異性糖抗原決定基(TA-Muc1)、VSIR (VISTA)及VTCN1。Non-limiting exemplary targets or immune cell engagers that antibodies can target include: Mullerian hormone receptor II (AMHR2), B7, B7H1, B7H2, B7H3, B7H4, BAFF-R, BCMA (B cell maturation antigen) , Bst1/CD157, C5 complement, CC chemokine receptor 4 (CCR4), CD123, CD137, CD19, CD20, CD25 (IL2RA), CD276, CD278, CD3, CD32, CD33, CD37, CD38, CD4 and HIV- 1 gp120 binding site, CD40, CD70, CD70 (member of the TNF receptor ligand family), CD80, CD86, claudin 18.2, c-MET, CSF1R, CTLA-4, EGFR, EGFR MET proto-oncogene , EPHA3, ERBB2, ERBB3, FGFR2b, FLT3, GITR, glucocorticoid-induced TNF receptor (GITR), HER2, HER3, HLA, ICOS, IDO1, IFNAR1, IFNAR2, IGF-1R, IL-3Rα (CD123), Respiratory fusion virus (RSV) recombinant of IL-5R, IL-5Rα, LAG-3, MET proto-oncogene, OX40 (CD134), PD-1, PD-L1, PD-L2, PVRIG, EBOV glycoprotein (GP) Glycosylated mucin-like domain, rhesus monkey (Rh) D, sialic acid immunoglobulin-like lectin 8 (Siglec-8), signaling lymphocyte activation molecule (SLAMF7/CS1), T cell receptor cytotoxic T Lymphocyte-associated antigen 4 (CTLA4), TIGIT, TIM3 (HAVCR2), tumor-specific carbohydrate epitope of Muc1 (TA-Muc1), VSIR (VISTA) and VTCN1.

在一些實施例中，抗體為免疫細胞接合子之促效劑。在一些此類實施例中，抗體為選自以下之免疫細胞接合子之促效劑：CD80、CD86、OX40 (CD134)、GITR、CD137、CD40、VTCN1、CD276、IFNAR2、IFNAR1、CSF1R、VSIR (VISTA)及HLA。In some embodiments, the antibody is an agonist of an immune cell conjugate. In some such embodiments, the antibody is an agonist of an immune cell engager selected from CD80, CD86, OX40 (CD134), GITR, CD137, CD40, VTCN1, CD276, IFNAR2, IFNAR1, CSF1R, VSIR ( VISTA) and HLA.

在一些實施例中，抗體為免疫細胞接合子之拮抗劑。在一些此類實施例中，抗體為選自以下之免疫細胞接合子之拮抗劑：CTLA-4、PD-1、PD-L1、PD-L2、LAG-3、B7、TIM3 (HAVCR2)、PVRIG、TIGIT、CD25 (IL2RA)及IDO1。In some embodiments, the antibody is an antagonist of an immune cell conjugate. In some such embodiments, the antibody is an antagonist of an immune cell engager selected from CTLA-4, PD-1, PD-L1, PD-L2, LAG-3, B7, TIM3 (HAVCR2), PVRIG , TIGIT, CD25 (IL2RA) and IDO1.

在某些實施例中，用於本文所提供之方法之結合免疫細胞接合子的抗體可為針對檢查點蛋白之抑制劑。在一些實施例中，用於本文所提供之方法之結合免疫細胞接合子之抗體可為PD-1抑制劑、PD-L1抑制劑、PD-L2抑制劑、CTLA-4抑制劑、LAG-3抑制劑、B7抑制劑、TIM3 (HAVCR2)抑制劑、OX40 (CD134)抑制劑、GITR促效劑、CD137促效劑，或CD40促效劑、VTCN1抑制劑、IDO1抑制劑、CD276抑制劑、PVRIG抑制劑、TIGIT抑制劑、CD25 (IL2RA)抑制劑、IFNAR2抑制劑、IFNAR1抑制劑、CSF1R抑制劑、VSIR (VISTA)抑制劑或靶向HLA之治療劑。此類抑制劑、活化劑或治療劑進一步提供於下文中。在本文中之任何實施例中，結合免疫細胞接合子之抗體可為包含增強與一或多種活化FcγR之結合的Fc的抗體。在一些實施例中，結合免疫細胞接合子之抗體為未經岩藻糖基化抗體。In certain embodiments, antibodies that bind immune cell engagers for use in the methods provided herein can be inhibitors of checkpoint proteins. In some embodiments, antibodies that bind immune cell engagers for use in the methods provided herein can be PD-1 inhibitors, PD-L1 inhibitors, PD-L2 inhibitors, CTLA-4 inhibitors, LAG-3 Inhibitor, B7 inhibitor, TIM3 (HAVCR2) inhibitor, OX40 (CD134) inhibitor, GITR agonist, CD137 agonist, or CD40 agonist, VTCN1 inhibitor, IDO1 inhibitor, CD276 inhibitor, PVRIG Inhibitors, TIGIT inhibitors, CD25 (IL2RA) inhibitors, IFNAR2 inhibitors, IFNAR1 inhibitors, CSF1R inhibitors, VSIR (VISTA) inhibitors, or HLA-targeted therapeutics. Such inhibitors, activators or therapeutics are further provided below. In any of the embodiments herein, the antibody that binds the immune cell adaptor can be an antibody comprising an Fc that enhances binding to one or more activating FcγRs. In some embodiments, the antibody that binds to the immune cell adaptor is an unfucosylated antibody.

在一些實施例中，結合免疫細胞接合子之抗體為CTLA-4抑制劑。在一個實施例中，CTLA-4抑制劑為抗CTLA-4抗體。抗CTLA-4抗體之實例包括但不限於描述於美國專利第5,811,097號、第5,811,097號、第5,855,887號、第6,051,227號、第6,207,157號、第6,682,736號、第6,984,720號及第7,605,238號中之彼等者，該等專利中之所有以其全文引用之方式併入本文中。在一個實施例中，抗-CTLA-4抗體為曲美單抗(tremelimumab) (亦稱為替西單抗(ticilimumab)或CP-675,206)或其未經岩藻糖基化版本。在另一實施例中，抗-CTLA-4抗體為伊匹單抗(ipilimumab) (亦稱為MDX-010或MDX-101)或其未經岩藻糖基化版本。伊匹單抗為結合於CTLA-4之完全人類單株IgG抗體。伊匹單抗以商標名Yervoy™出售。In some embodiments, the antibody that binds to the immune cell engager is a CTLA-4 inhibitor. In one embodiment, the CTLA-4 inhibitor is an anti-CTLA-4 antibody. Examples of anti-CTLA-4 antibodies include, but are not limited to, those described in US Pat. Nos. 5,811,097, 5,811,097, 5,855,887, 6,051,227, 6,207,157, 6,682,736, 6,984,720, and 7,605,238 , all of these patents are incorporated herein by reference in their entirety. In one embodiment, the anti-CTLA-4 antibody is tremelimumab (also known as ticilimumab or CP-675,206) or a non-fucosylated version thereof. In another embodiment, the anti-CTLA-4 antibody is ipilimumab (also known as MDX-010 or MDX-101) or an unfucosylated version thereof. Ipilimumab is a fully human monoclonal IgG antibody that binds to CTLA-4. Ipilimumab is sold under the brand name Yervoy™.

在某些實施例中，結合免疫細胞接合子之抗體為PD-1/PD-L1抑制劑。PD-l/PD-L1抑制劑之實例包括但不限於美國專利第7,488,802號、第7,943,743號、第8,008,449號、第8,168,757號、第8,217,149號，以及PCT專利申請公開案第WO2003042402號、第WO2008156712號、第WO2010089411號、第WO2010036959號、第WO2011066342號、第WO2011159877號、第WO2011082400號及第WO2011161699中所描述之PD-l/PD-L1抑制劑，所有文獻以全文引用之方式併入本文中。In certain embodiments, the antibody that binds to the immune cell engager is a PD-1/PD-L1 inhibitor. Examples of PD-1/PD-L1 inhibitors include, but are not limited to, US Patent Nos. 7,488,802, 7,943,743, 8,008,449, 8,168,757, 8,217,149, and PCT Patent Application Publication Nos. WO2003042402, WO2008156712 , WO2010089411, WO2010036959, WO2011066342, WO2011159877, WO2011082400 and WO2011161699, all of which are incorporated herein by reference in their entirety.

在一些實施例中，結合免疫細胞接合子之抗體為PD-1抑制劑。在一個實施例中，PD-1抑制劑為抗PD-1抗體。在一個實施例中，抗PD-1抗體為BGB-A317、納武單抗(nivolumab) (亦被稱作ONO-4538、BMS-936558或MDX1106)或派立珠單抗(pembrolizumab) (亦被稱作MK-3475、SCH 900475或拉立珠單抗(lambrolizumab))或其未經岩藻糖基化版本。在一個實施例中，抗PD-1抗體為納武單抗或其未經岩藻糖基化版本。納武單抗為人類IgG4抗PD-1單株抗體，且以商標名Opdivo™出售。在另一實施例中，抗PD-1抗體為派立珠單抗或其未經岩藻糖基化版本。派立珠單抗為人類化單株IgG4抗體且以商標名Keytruda™出售。在又一實施例中，抗PD-1抗體為CT-011、人類化抗體或其未經岩藻糖基化版本。在復發情況下單獨投與CT-011未能展示治療急性骨髓白血病(AML)之反應。在又一實施例中，抗PD-1抗體為AMP-224、融合蛋白或其未經岩藻糖基化版本。在另一實施例中，PD-1抗體為BGB-A317或其未經岩藻糖基化版本。BGB-A317為一種單株抗體，其中特異性地工程改造出結合Fc γ受體I之能力，且其具有以高親和力及優良目標特異性與PD-1獨特結合之特徵。在一個實施例中，PD-1抗體為西米普利單抗(cemiplimab)或其未經岩藻糖基化版本。在另一實施例中，PD-1抗體為卡瑞利珠單抗(camrelizumab)或其未經岩藻糖基化版本。在另一實施例中，PD-1抗體為斯迪利單抗(sintilimab)或其未經岩藻糖基化版本。在一個實施例中，PD-1抗體為替雷利珠單抗(tislelizumab)或其未經岩藻糖基化版本。在某些實施例中，PD-1抗體為TSR-042或其未經岩藻糖基化版本。在又一實施例中，PD-1抗體為PDR001或其未經岩藻糖基化版本。在又一實施例中，PD-1抗體為特瑞普利單抗(toripalimab)或其未經岩藻糖基化版本。In some embodiments, the antibody that binds to the immune cell engager is a PD-1 inhibitor. In one embodiment, the PD-1 inhibitor is an anti-PD-1 antibody. In one embodiment, the anti-PD-1 antibody is BGB-A317, nivolumab (also known as ONO-4538, BMS-936558 or MDX1106) or pembrolizumab (also known as Known as MK-3475, SCH 900475 or lambrolizumab) or its unfucosylated version. In one embodiment, the anti-PD-1 antibody is nivolumab or an unfucosylated version thereof. Nivolumab is a human IgG4 anti-PD-1 monoclonal antibody and is sold under the brand name Opdivo™. In another embodiment, the anti-PD-1 antibody is pelivizumab or a non-fucosylated version thereof. Perilizumab is a humanized monoclonal IgG4 antibody and is sold under the brand name Keytruda™. In yet another embodiment, the anti-PD-1 antibody is CT-011, a humanized antibody, or a non-fucosylated version thereof. Administration of CT-011 alone failed to demonstrate response to treatment of acute myeloid leukemia (AML) in the setting of relapse. In yet another embodiment, the anti-PD-1 antibody is AMP-224, a fusion protein or an unfucosylated version thereof. In another embodiment, the PD-1 antibody is BGB-A317 or a non-fucosylated version thereof. BGB-A317 is a monoclonal antibody specifically engineered for the ability to bind Fcγ receptor I, and which is characterized by unique binding to PD-1 with high affinity and excellent target specificity. In one embodiment, the PD-1 antibody is cemiplimab or an unfucosylated version thereof. In another embodiment, the PD-1 antibody is camrelizumab or an unfucosylated version thereof. In another embodiment, the PD-1 antibody is sintilimab or a non-fucosylated version thereof. In one embodiment, the PD-1 antibody is tislelizumab or an unfucosylated version thereof. In certain embodiments, the PD-1 antibody is TSR-042 or an unfucosylated version thereof. In yet another embodiment, the PD-1 antibody is PDR001 or a non-fucosylated version thereof. In yet another embodiment, the PD-1 antibody is toripalimab or a non-fucosylated version thereof.

在某些實施例中，結合免疫細胞接合子之抗體為PD-L1抑制劑。在一個實施例中，PD-L1抑制劑為抗PD-L1抗體。在一個實施例中，抗PD-L1抗體為MEDI4736 (德瓦魯單抗(durvalumab))或其未經岩藻糖基化版本。在另一實施例中，抗PD-L1抗體為BMS-936559 (亦稱為MDX-1105-01)或其未經岩藻糖基化版本。在又一實施例中，PD-L1抑制劑為阿特珠單抗(atezolizumab) (亦稱為MPDL3280A及Tecentriq®)或其未經岩藻糖基化版本。在另一實施例中，PD-L1抑制劑為阿維魯單抗(avelumab)或其未經岩藻糖基化版本。In certain embodiments, the antibody that binds to the immune cell engager is a PD-L1 inhibitor. In one embodiment, the PD-L1 inhibitor is an anti-PD-L1 antibody. In one embodiment, the anti-PD-L1 antibody is MEDI4736 (durvalumab) or an unfucosylated version thereof. In another embodiment, the anti-PD-L1 antibody is BMS-936559 (also known as MDX-1105-01) or a non-fucosylated version thereof. In yet another embodiment, the PD-L1 inhibitor is atezolizumab (also known as MPDL3280A and Tecentriq®) or a non-fucosylated version thereof. In another embodiment, the PD-L1 inhibitor is avelumab or a non-fucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為PD-L2抑制劑。在一個實施例中，PD-L2抑制劑為抗PD-L2抗體。在一個實施例中，抗PD-L2抗體為rHIgM12B7A或其未經岩藻糖基化版本。In one embodiment, the antibody that binds to the immune cell engager is a PD-L2 inhibitor. In one embodiment, the PD-L2 inhibitor is an anti-PD-L2 antibody. In one embodiment, the anti-PD-L2 antibody is rHIgM12B7A or an unfucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為淋巴球活化基因-3 (LAG-3)抑制劑。在一個實施例中，LAG-3抑制劑為IMP321(一種可溶性Ig融合蛋白) (Brignone等人， J. Immunol., 2007, 179, 4202-4211)，或其未經岩藻糖基化版本。在另一實施例中，LAG-3抑制劑為BMS-986016或其未經岩藻糖基化版本。 In one embodiment, the antibody that binds to the immune cell engager is a lymphocyte activation gene-3 (LAG-3) inhibitor. In one embodiment, the LAG-3 inhibitor is IMP321, a soluble Ig fusion protein (Brignone et al., J. Immunol. , 2007 , 179, 4202-4211), or an unfucosylated version thereof. In another embodiment, the LAG-3 inhibitor is BMS-986016 or a non-fucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為B7抑制劑。在一個實施例中，B7抑制劑為B7-H3抑制劑或B7-H4抑制劑。在一個實施例中，B7-H3抑制劑為MGA271，抗B7-H3抗體(Loo等人， Clin. Cancer Res., 2012, 3834)，或其未經岩藻糖基化版本。在一些實施例中，B7抑制劑為B7-H4抑制劑。非限制性例示性B7-H4抑制劑為FPA150(一種針對B7-H4之未經岩藻糖基化抗體)。 參見PCT/US2018/047805。 In one embodiment, the antibody that binds to the immune cell engager is a B7 inhibitor. In one embodiment, the B7 inhibitor is a B7-H3 inhibitor or a B7-H4 inhibitor. In one embodiment, the B7-H3 inhibitor is MGA271, an anti-B7-H3 antibody (Loo et al., Clin. Cancer Res. , 2012 , 3834), or an unfucosylated version thereof. In some embodiments, the B7 inhibitor is a B7-H4 inhibitor. A non-limiting exemplary B7-H4 inhibitor is FPA150, a non-fucosylated antibody directed against B7-H4. See PCT/US2018/047805.

在一個實施例中，結合免疫細胞接合子之抗體為TIM3 (T細胞免疫球蛋白域及黏蛋白域3)抑制劑(Fourcade等人, J. Exp. Med., 2010, 207, 2175-86；Sakuishi等人, J. Exp. Med., 2010, 207, 2187-94)。 In one embodiment, the antibody that binds the immune cell engager is a TIM3 (T cell immunoglobulin domain and mucin domain 3) inhibitor (Fourcade et al., J. Exp. Med. , 2010 , 207, 2175-86; Sakuishi et al, J. Exp. Med. , 2010 , 207, 2187-94).

在一個實施例中，結合免疫細胞接合子之抗體為OX40 (CD134)促效劑抗體。在某些實施例中，抗OX40抗體為MEDI6469或其未經岩藻糖基化版本。In one embodiment, the antibody that binds the immune cell engager is an OX40 (CD134) agonist antibody. In certain embodiments, the anti-OX40 antibody is MEDI6469 or an unfucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為GITR促效劑。在一個實施例中，免疫細胞接合子為抗GITR抗體或其未經岩藻糖基化版本。在一個實施例中，抗GITR抗體為TRX518或其未經岩藻糖基化版本。In one embodiment, the antibody that binds to the immune cell engager is a GITR agonist. In one embodiment, the immune cell adaptor is an anti-GITR antibody or an unfucosylated version thereof. In one embodiment, the anti-GITR antibody is TRX518 or an unfucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為CD137促效劑。在一個實施例中，免疫細胞接合子為抗CD137抗體。在一個實施例中，抗CD137抗體為烏瑞魯單抗(urelumab)或其未經岩藻糖基化版本。在另一實施例中，抗CD137抗體為PF-05082566或其未經岩藻糖基化版本。In one embodiment, the antibody that binds to the immune cell engager is a CD137 agonist. In one embodiment, the immune cell adaptor is an anti-CD137 antibody. In one embodiment, the anti-CD137 antibody is urelumab or an unfucosylated version thereof. In another embodiment, the anti-CD137 antibody is PF-05082566 or a non-fucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為CD40促效劑。在一個實施例中，結合免疫細胞接合子之抗體為抗CD40抗體。在一個實施例中，抗CD40抗體為CF-870,893或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為MP0317 (Molecular Partners)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為YH003 (Eucure Biopharma)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為CDX-1140 (Celldex Therapeutics)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為YH003 (Eucure Biopharma)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為米佐利單抗(mitazalimab) (Alligator Bioscience)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為ABBV-927 (AbbVie)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為索替利單抗(sotigalimab) (Apexigen)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為GEN1042 (Genmab)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為2141 V-11 (Rockefeller University)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為塞立路單抗(selicrelumab) (Roche)或其未經岩藻糖基化版本。在一個實施例中，抗CD40抗體為SEA-CD40 (Seagen)，其為鼠類S2C6之未經岩藻糖基化人類化版本且其包含分別包含SEQ ID NO: 30至35之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。對應VH及VL分別包含SEQ ID NO: 28及29之胺基酸序列。SEA-CD40描述於美國專利公開案第2017/0333556號及第2017/0137528號中，該兩件公開案以引用之方式併入本文中。In one embodiment, the antibody that binds to the immune cell engager is a CD40 agonist. In one embodiment, the antibody that binds to the immune cell adaptor is an anti-CD40 antibody. In one embodiment, the anti-CD40 antibody is CF-870,893 or an unfucosylated version thereof. In one embodiment, the anti-CD40 antibody is MP0317 (Molecular Partners) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is YH003 (Eucure Biopharma) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is CDX-1140 (Celldex Therapeutics) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is YH003 (Eucure Biopharma) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is mitazalimab (Alligator Bioscience) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is ABBV-927 (AbbVie) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is sotigalimab (Apexigen) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is GEN1042 (Genmab) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is 2141 V-11 (Rockefeller University) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is selicrelumab (Roche) or a non-fucosylated version thereof. In one embodiment, the anti-CD40 antibody is SEA-CD40 (Seagen), which is a non-fucosylated humanized version of murine S2C6 and which comprises the amino acid sequences comprising SEQ ID NOs: 30 to 35, respectively The heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3. The corresponding VH and VL comprise the amino acid sequences of SEQ ID NOs: 28 and 29, respectively. SEA-CD40 is described in US Patent Publication Nos. 2017/0333556 and 2017/0137528, both of which are incorporated herein by reference.

在一些實施例中，結合免疫細胞接合子之抗體為結合CD70之抗體。在一些實施例中，抗體為SEA-CD70。 參見例如美國專利案第8,067,546號；本文中之序列表。 In some embodiments, the antibody that binds to the immune cell engager is an antibody that binds CD70. In some embodiments, the antibody is SEA-CD70. See , eg, US Patent No. 8,067,546; Sequence Listing herein.

在一些實施例中，結合免疫細胞接合子之抗體為結合BCMA之抗體。在一些實施例中，抗體為SEA-BCMA。 參見例如美國公開案第2017/0233484號及WO 2017/143069 (分別為SEQ ID NO: 13及19之VH及VL；SEQ ID NO: 60、61、62、90、91、92之CDR， 參見美國公開案第2017/0233484號)； 亦參見本文中之序列表(分別為SEQ ID NO: 45及46之VH及VL；SEQ ID NO: 47至52之CDR)。 In some embodiments, the antibody that binds to the immune cell engager is an antibody that binds BCMA. In some embodiments, the antibody is SEA-BCMA. See, eg , U.S. Publication No. 2017/0233484 and WO 2017/143069 (VH and VL of SEQ ID NOs: 13 and 19, respectively; CDRs of SEQ ID NOs: 60, 61, 62, 90, 91, 92, see U.S. Publication No. 2017/0233484); see also the Sequence Listing herein (VH and VL of SEQ ID NOs: 45 and 46, respectively; CDRs of SEQ ID NOs: 47-52).

在一個實施例中，結合免疫細胞接合子之抗體為抗介白素-15抗體。In one embodiment, the antibody that binds to the immune cell adaptor is an anti-interleukin-15 antibody.

在一個實施例中，結合免疫細胞接合子之抗體為VTCN抑制劑。在一個實施例中，VTCN抑制劑為FPA150或其未經岩藻糖基化版本。In one embodiment, the antibody that binds to the immune cell engager is a VTCN inhibitor. In one embodiment, the VTCN inhibitor is FPA150 or an unfucosylated version thereof.

在一個實施例中，結合免疫細胞接合子之抗體為抗IDO拮抗劑抗體。在一些實施例中，結合免疫細胞接合子之抗體為TIGIT抑制劑。在某些實施例中，TIGIT抑制劑為抗TIGIT抗體。在一個實施例中，TIGIT抑制劑為MTIG7192A或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為BMS-986207 (BMS)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為OMP-313M32或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為MK-7684。在另一實施例中，TIGIT抑制劑為AB154或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為CGEN-15137或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為SEA-TGT。在另一實施例中，TIGIT抑制劑為ASP8374 (Astellas)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為AJUD008或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為AB308 (Arcus Biosciences)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為AGEN1327 (Agenus)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為AK127 (Akeso Biopharma)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為BAT6005 (Bio-Thera Solutions)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為BAT6021 (Bio-Thera Solutions)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為CASC-674 (Seagen)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為COM902 (Compugen)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為多瓦利單抗(domvanalimab)(Arcus Biosciences) 或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為厄提吉利單抗(etigilimab) (Mereo BioPharma)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為GSK4428859 (GSK)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為HL186 (HanAll Biopharma)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為MIL-100 (Beijing Mabworks Biotech)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為YH-29143 (Yu Han)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為HLX53 (Shanghai Henlius Biotech)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為IBI939 (Innovent Biologics)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為JS006 (Junshi Biosciences)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為M6223 (Merck KGaA)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為MG1131 (Mogam Institute)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為西培利單抗(ociperlimab) (BeiGene)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為替拉格魯單抗(tiragolumab) (Roche；描述於美國專利案第10,047,158號中)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為TJT6 (I-Mab Biopharma)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為韋博妥單抗(vibostolimab) (MSD；描述於美國專利案第10,618,958號中)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為YBL-012 (Y Biologics)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為IBI-939 (Innovent)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為AZD2936 (AstraZeneca)或其未經岩藻糖基化版本。在一個實施例中，TIGIT抑制劑為EOS-448 (iTeos/GSK)或其未經岩藻糖基化版本。在另一實施例中，TIGIT抑制劑為BAT6005 (Bio-Thera)或其未經岩藻糖基化版本。在又一實施例中，TIGIT抑制劑為AGEN1777 (BMS/Agenus)或其未經岩藻糖基化版本。In one embodiment, the antibody that binds the immune cell engager is an anti-IDO antagonist antibody. In some embodiments, the antibody that binds the immune cell engager is a TIGIT inhibitor. In certain embodiments, the TIGIT inhibitor is an anti-TIGIT antibody. In one embodiment, the TIGIT inhibitor is MTIG7192A or an unfucosylated version thereof. In another embodiment, the TIGIT inhibitor is BMS-986207 (BMS) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is OMP-313M32 or an unfucosylated version thereof. In one embodiment, the TIGIT inhibitor is MK-7684. In another embodiment, the TIGIT inhibitor is AB154 or an unfucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is CGEN-15137 or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is SEA-TGT. In another embodiment, the TIGIT inhibitor is ASP8374 (Astellas) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is AJUD008 or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is AB308 (Arcus Biosciences) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is AGEN1327 (Agenus) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is AK127 (Akeso Biopharma) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is BAT6005 (Bio-Thera Solutions) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is BAT6021 (Bio-Thera Solutions) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is CASC-674 (Seagen) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is COM902 (Compugen) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is domvanalimab (Arcus Biosciences) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is etigilimab (Mereo BioPharma) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is GSK4428859 (GSK) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is HL186 (HanAll Biopharma) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is MIL-100 (Beijing Mabworks Biotech) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is YH-29143 (Yu Han) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is HLX53 (Shanghai Henlius Biotech) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is IBI939 (Innovent Biologics) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is JS006 (Junshi Biosciences) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is M6223 (Merck KGaA) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is MG1131 (Mogam Institute) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is ociperlimab (BeiGene) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is tiragolumab (Roche; described in US Pat. No. 10,047,158) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is TJT6 (I-Mab Biopharma) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is vibostolimab (MSD; described in US Pat. No. 10,618,958) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is YBL-012 (Y Biologics) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is IBI-939 (Innovent) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is AZD2936 (AstraZeneca) or a non-fucosylated version thereof. In one embodiment, the TIGIT inhibitor is EOS-448 (iTeos/GSK) or a non-fucosylated version thereof. In another embodiment, the TIGIT inhibitor is BAT6005 (Bio-Thera) or a non-fucosylated version thereof. In yet another embodiment, the TIGIT inhibitor is AGEN1777 (BMS/Agenus) or a non-fucosylated version thereof.

在一些實施例中，結合免疫細胞接合子之抗體為VSIR抑制劑。在某些實施例中，VSIR抑制劑為抗VSIR抗體。在一個實施例中，VSIR抑制劑為MTIG7192A或其未經岩藻糖基化版本。在另一實施例中，VSIR抑制劑為CA-170或其未經岩藻糖基化版本。在又一實施例中，VSIR抑制劑為JNJ 61610588或其未經岩藻糖基化版本。在一個實施例中，VSIR抑制劑為HMBD-002或其未經岩藻糖基化版本。In some embodiments, the antibody that binds to the immune cell engager is a VSIR inhibitor. In certain embodiments, the VSIR inhibitor is an anti-VSIR antibody. In one embodiment, the VSIR inhibitor is MTIG7192A or an unfucosylated version thereof. In another embodiment, the VSIR inhibitor is CA-170 or an unfucosylated version thereof. In yet another embodiment, the VSIR inhibitor is JNJ 61610588 or a non-fucosylated version thereof. In one embodiment, the VSIR inhibitor is HMBD-002 or an unfucosylated version thereof.

在一些實施例中，結合免疫細胞接合子之抗體為TIM3抑制劑。在某些實施例中，TIM3抑制劑為抗TIM3抗體。在一個實施例中，TIM3抑制劑為AJUD009或其未經岩藻糖基化版本。In some embodiments, the antibody that binds to the immune cell engager is a TIM3 inhibitor. In certain embodiments, the TIM3 inhibitor is an anti-TIM3 antibody. In one embodiment, the TIM3 inhibitor is AJUD009 or an unfucosylated version thereof.

在一些實施例中，結合免疫細胞接合子之抗體為CD25 (IL2RA)抑制劑。在某些實施例中，CD25 (IL2RA)抑制劑為抗CD25 (IL2RA)抗體。在一個實施例中，CD25 (IL2RA)抑制劑為達利珠單抗(daclizumab)或其未經岩藻糖基化版本。在另一實施例中，CD25 (IL2RA)抑制劑為巴利昔單抗(basiliximab)或其未經岩藻糖基化版本。In some embodiments, the antibody that binds to the immune cell engager is a CD25 (IL2RA) inhibitor. In certain embodiments, the CD25 (IL2RA) inhibitor is an anti-CD25 (IL2RA) antibody. In one embodiment, the CD25 (IL2RA) inhibitor is daclizumab or an unfucosylated version thereof. In another embodiment, the CD25 (IL2RA) inhibitor is basiliximab or an unfucosylated version thereof.

在一些實施例中，結合免疫細胞接合子之抗體為IFNAR1抑制劑。在某些實施例中，IFNAR1抑制劑為抗IFNAR1抗體。在一個實施例中，IFNAR1抑制劑為阿尼富路單抗(anifrolumab)或其未經岩藻糖基化版本。在另一實施例中，IFNAR1抑制劑為絲法力單抗(sifalimumab)或其未經岩藻糖基化版本。In some embodiments, the antibody that binds to the immune cell engager is an IFNAR1 inhibitor. In certain embodiments, the IFNAR1 inhibitor is an anti-IFNAR1 antibody. In one embodiment, the IFNAR1 inhibitor is anifrolumab or an unfucosylated version thereof. In another embodiment, the IFNAR1 inhibitor is sifalimumab or a non-fucosylated version thereof.

在一些實施例中，結合免疫細胞接合子之抗體為CSF1R抑制劑。在某些實施例中，CSF1R抑制劑為抗CSF1R抗體。在一個實施例中，CSF1R抑制劑為吡昔替尼(pexidartinib)或其未經岩藻糖基化版本。在另一實施例中，CSF1R抑制劑為艾瑪圖單抗(emactuzumab)或其未經岩藻糖基化版本。在又一實施例中，CSF1R抑制劑為卡比利珠單抗(cabiralizumab)或其未經岩藻糖基化版本。在一個實施例中，CSF1R抑制劑為ARRY-382或其未經岩藻糖基化版本。在另一實施例中，CSF1R抑制劑為BLZ945或其未經岩藻糖基化版本。在又一實施例中，CSF1R抑制劑為AJUD010或其未經岩藻糖基化版本。在一個實施例中，CSF1R抑制劑為AMG820或其未經岩藻糖基化版本。在另一實施例中，CSF1R抑制劑為IMC-CS4或其未經岩藻糖基化版本。在又一實施例中，CSF1R抑制劑為JNJ-40346527或其未經岩藻糖基化版本。在一個實施例中，CSF1R抑制劑為PLX5622或其未經岩藻糖基化版本。在另一實施例中，CSF1R抑制劑為FPA008或其未經岩藻糖基化版本。In some embodiments, the antibody that binds to the immune cell engager is a CSF1R inhibitor. In certain embodiments, the CSF1R inhibitor is an anti-CSF1R antibody. In one embodiment, the CSF1R inhibitor is pexidartinib or an unfucosylated version thereof. In another embodiment, the CSF1R inhibitor is emactuzumab or an unfucosylated version thereof. In yet another embodiment, the CSF1R inhibitor is cabiralizumab or an unfucosylated version thereof. In one embodiment, the CSF1R inhibitor is ARRY-382 or an unfucosylated version thereof. In another embodiment, the CSF1R inhibitor is BLZ945 or an unfucosylated version thereof. In yet another embodiment, the CSF1R inhibitor is AJUD010 or a non-fucosylated version thereof. In one embodiment, the CSF1R inhibitor is AMG820 or an unfucosylated version thereof. In another embodiment, the CSF1R inhibitor is IMC-CS4 or an unfucosylated version thereof. In yet another embodiment, the CSF1R inhibitor is JNJ-40346527 or a non-fucosylated version thereof. In one embodiment, the CSF1R inhibitor is PLX5622 or a non-fucosylated version thereof. In another embodiment, the CSF1R inhibitor is FPA008 or an unfucosylated version thereof.

在各種實施例中,結合免疫細胞接合子之抗體具有呈任何組合之以下活性中之一或多者或所有：1)耗盡T調節性(Treg)細胞，2)活化抗原呈遞細胞(APC)，3)增強CD8 T細胞反應，4)上調共刺激受體及/或5)促進免疫活化細胞介素(諸如CXCL10及/或IFNγ)之釋放。在一些實施例中，結合免疫細胞接合子之抗體促使免疫活化細胞介素(例如CXCL10及IFNγ)之釋放程度大於免疫抑制細胞介素(諸如IL10及/或MDC)之釋放。 B. 例示性抗 TIGIT 抗體 In various embodiments, the antibody that binds the immune cell engager has one or more or all of the following activities in any combination: 1) deplete T regulatory (Treg) cells, 2) activate antigen presenting cells (APCs) , 3) enhance CD8 T cell responses, 4) upregulate co-stimulatory receptors and/or 5) promote release of immune activating interferons such as CXCL10 and/or IFNγ. In some embodiments, antibodies that bind to immune cell engagers promote the release of immune activating interleukins (eg, CXCL10 and IFNy) to a greater extent than immunosuppressive interleukins (eg, IL10 and/or MDC). B. Exemplary Anti- TIGIT Antibodies

在一個態樣中，結合於人類TIGIT (具有Ig域及ITIM域之T細胞免疫受體)之抗體經提供為針對免疫細胞接合子之抗體。如本文中所描述，在一些實施例中，抗TIGIT抗體抑制TIGIT與配位體CD155及CD112中之一或兩者之間的相互作用。在一些實施例中，抗TIGIT抗體在功能性生物分析中抑制TIGIT與CD155之間的相互作用，允許發生CD155-CD226傳信。在一些實施例中，抗TIGIT抗體展現與抗PD-1藥劑(例如抗PD-1抗體)或抗PD-L1藥劑(例如抗PD-L1抗體)之協同作用。在一些實施例中，用於本發明方法之抗TIGIT抗體為SEA-TGT，其為包含分別包含SEQ ID NO: 7、10、14、17、18、及19之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3的未經岩藻糖基化IgG1抗體。對應VH及VL分別包含SEQ ID NO: 1及6之胺基酸序列。In one aspect, an antibody that binds to human TIGIT (a T cell immune receptor with Ig and ITIM domains) is provided as an antibody against an immune cell engager. As described herein, in some embodiments, an anti-TIGIT antibody inhibits the interaction of TIGIT with one or both of the ligands CD155 and CD112. In some embodiments, the anti-TIGIT antibody inhibits the interaction between TIGIT and CD155 in a functional biological assay, allowing CD155-CD226 signaling to occur. In some embodiments, the anti-TIGIT antibody exhibits synergy with an anti-PD-1 agent (eg, an anti-PD-1 antibody) or an anti-PD-L1 agent (eg, an anti-PD-L1 antibody). In some embodiments, the anti-TIGIT antibody used in the methods of the invention is SEA-TGT, which is a heavy chain CDR1 comprising the amino acid sequences comprising SEQ ID NOs: 7, 10, 14, 17, 18, and 19, respectively , CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 unfucosylated IgG1 antibody. The corresponding VH and VL comprise the amino acid sequences of SEQ ID NOs: 1 and 6, respectively.

本發明人出人意料地發現，具有諸如可藉由未經岩藻糖基化IgG1抗體達成之增強之效應功能的抗TIGIT抗體耗乏Treg細胞且展示改良之活體內功效。因此，在各種實施例中，提供未經岩藻糖基化抗TIGIT抗體。The inventors have unexpectedly discovered that anti-TIGIT antibodies with enhanced effector functions such as can be achieved by non-fucosylated IgGl antibodies deplete Treg cells and exhibit improved in vivo efficacy. Thus, in various embodiments, non-fucosylated anti-TIGIT antibodies are provided.

在一些實施例中，諸如未經岩藻糖基化抗TIGIT抗體之抗TIGIT抗體以高親和力結合於人類TIGIT蛋白質(SEQ ID NO:218)或其部分。在一些實施例中，抗體對於人類TIGIT之結合親和力(K D)小於5 nM、小於1 nM、小於500 pM、小於250 pM、小於150 pM、小於100 pM、小於50 pM、小於40 pM、小於30 pM、小於20 pM或小於約10 pM。在一些實施例中，抗體對於人類TIGIT之結合親和力(K D)小於50 pM。在一些實施例中，抗體對於人類TIGIT之K D在約1 pM至約5 nM，例如約1 pM至約1 nM、約1 pM至約500 pM、約5 pM至約250 pM或約10 pM至約100 pM範圍內。 In some embodiments, an anti-TIGIT antibody, such as a non-fucosylated anti-TIGIT antibody, binds with high affinity to human TIGIT protein (SEQ ID NO: 218) or a portion thereof. In some embodiments, the antibody has a binding affinity (K D ) for human TIGIT of less than 5 nM, less than 1 nM, less than 500 pM, less than 250 pM, less than 150 pM, less than 100 pM, less than 50 pM, less than 40 pM, less than 30 pM, less than 20 pM, or less than about 10 pM. In some embodiments, the antibody has a binding affinity (K D ) for human TIGIT of less than 50 pM. In some embodiments, the antibody has a KD for human TIGIT of about 1 pM to about 5 nM, eg, about 1 pM to about 1 nM, about 1 pM to about 500 pM, about 5 pM to about 250 pM, or about 10 pM to the range of about 100 pM.

在一些實施例中，除了以高親和力結合於人類TIGIT之外，未經岩藻糖基化抗TIGIT抗體展現與食蟹獼猴(「cyno」) TIGIT及/或小鼠TIGIT之交叉反應性。在一些實施例中，抗TIGIT抗體以100 nM或更小之結合親和力(K D)結合於小鼠TIGIT。在一些實施例中，抗TIGIT抗體以5 nM或更小之K D結合於人類TIGIT，且以100 nM或更小之K D與小鼠TIGIT交叉反應。在一些實施例中，結合於人類TIGIT之抗TIGIT抗體亦展現與食蟹獼猴TIGIT及小鼠TIGIT兩者之交叉反應性。 In some embodiments, in addition to binding to human TIGIT with high affinity, the non-fucosylated anti-TIGIT antibody exhibits cross-reactivity with cynomolgus monkey ("cyno") TIGIT and/or mouse TIGIT. In some embodiments, the anti-TIGIT antibody binds to mouse TIGIT with a binding affinity (K D ) of 100 nM or less. In some embodiments, the anti-TIGIT antibody binds to human TIGIT with a K of 5 nM or less, and cross-reacts with mouse TIGIT with a K of 100 nM or less. In some embodiments, anti-TIGIT antibodies that bind to human TIGIT also exhibit cross-reactivity with both cynomolgus monkey TIGIT and mouse TIGIT.

在一些實施例中，抗體交叉反應性係藉由偵測抗TIGIT抗體與表現於細胞(例如表現人類TIGIT、食蟹獼猴TIGIT或小鼠TIGIT之細胞株或內源性表現TIGIT的原代細胞，例如內源性表現人類TIGIT、cyno TIGIT或小鼠TIGIT之原代T細胞)上之TIGIT的特異性結合來測定。在一些實施例中，抗體結合及抗體交叉反應性係藉由偵測抗TIGIT抗體與純化或重組TIGIT (例如純化或重組人類TIGIT、純化或重組cyno TIGIT或純化或重組小鼠TIGIT)或包含TIGIT之嵌合蛋白(例如包含人類TIGIT、食蟹獼猴TIGIT或小鼠TIGIT之Fc融合蛋白或包含人類TIGIT、cyno TIGIT或小鼠TIGIT之His標記之蛋白質)的特異性結合來測定。 In some embodiments, antibody cross-reactivity is achieved by detecting anti-TIGIT antibodies with cell lines expressing human TIGIT, cynomolgus monkey TIGIT, or mouse TIGIT, or primary cells endogenously expressing TIGIT, For example, specific binding of TIGIT on primary T cells expressing human TIGIT, cyno TIGIT or mouse TIGIT endogenously is determined. In some embodiments, antibody binding and antibody cross-reactivity are achieved by detecting anti-TIGIT antibodies with purified or recombinant TIGIT (eg, purified or recombinant human TIGIT, purified or recombinant cyno TIGIT, or purified or recombinant mouse TIGIT) or comprising TIGIT The specific binding of a chimeric protein such as an Fc fusion protein comprising human TIGIT, cynomolgus TIGIT or mouse TIGIT or a His-tagged protein comprising human TIGIT, cyno TIGIT or mouse TIGIT was determined.

在一些實施例中，本文提供之抗TIGIT抗體抑制TIGIT與配位體CD155之間的相互作用。在一些實施例中，本文提供之抗TIGIT抗體抑制TIGIT與配位體CD112之間的相互作用。在一些實施例中，本文提供之抗TIGIT抗體抑制TIGIT與配位體CD155及CD112兩者之間的相互作用。In some embodiments, the anti-TIGIT antibodies provided herein inhibit the interaction between TIGIT and the ligand CD155. In some embodiments, the anti-TIGIT antibodies provided herein inhibit the interaction between TIGIT and the ligand CD112. In some embodiments, the anti-TIGIT antibodies provided herein inhibit the interaction between TIGIT and both the ligands CD155 and CD112.

在一些實施例中，結合於人類TIGIT之抗TIGIT抗體包含衍生於本文所描述之以下抗體中之任一者的輕鏈可變區序列或其部分及/或重鏈可變區序列或其部分：純系13、純系13A、純系13B、純系13C或純系13D。抗TIGIT抗體純系13、純系13A、純系13B、純系13C及純系13D之CDR、輕鏈可變域(VL)及重鏈可變域(VH)的胺基酸序列闡述於以下序列表中。In some embodiments, an anti-TIGIT antibody that binds to human TIGIT comprises a light chain variable region sequence or portion thereof and/or a heavy chain variable region sequence or portion thereof derived from any of the following antibodies described herein : Pure line 13, pure line 13A, pure line 13B, pure line 13C or pure line 13D. The amino acid sequences of the CDRs, light chain variable domains (VL) and heavy chain variable domains (VH) of the anti-TIGIT antibodies clone 13, clone 13A, clone 13B, clone 13C and clone 13D are set forth in the Sequence Listing below.

在一些實施例中，抗TIGIT抗體包含以下中之一或多者(例如一者、兩者、三者、四者、五者或六者)： 重鏈CDR1序列，其包含選自SEQ ID NO:7、SEQ ID NO:8及SEQ ID NO:9之胺基酸序列； 重鏈CDR2序列，其包含選自SEQ ID NO:10、SEQ ID NO:11、SEQ ID NO:12及SEQ ID NO:13之胺基酸序列； 重鏈CDR3序列，其包含選自SEQ ID NO:14、SEQ ID NO:15及16之胺基酸序列； 輕鏈CDR1序列，其包含SEQ ID NO:17之胺基酸序列； 輕鏈CDR2序列，其包含SEQ ID NO:18之胺基酸序列；及/或 輕鏈CDR3序列，其包含SEQ ID NO:19之胺基酸序列。 In some embodiments, the anti-TIGIT antibody comprises one or more of the following (eg, one, two, three, four, five, or six): A heavy chain CDR1 sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO:7, SEQ ID NO:8 and SEQ ID NO:9; a heavy chain CDR2 sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12 and SEQ ID NO: 13; a heavy chain CDR3 sequence comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 14, SEQ ID NO: 15 and 16; A light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 17; A light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 18; and/or A light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:19.

在一些實施例中，抗TIGIT抗體包含：重鏈CDR1序列，其包含SEQ ID NO: 7、SEQ ID NO: 8或SEQ ID NO: 9之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO: 10、SEQ ID NO: 11、SEQ ID NO: 12或SEQ ID NO: 13之胺基酸序列；以及重鏈CDR3序列，其包含SEQ ID NO: 14、SEQ ID NO: 15或16之胺基酸序列。 In some embodiments, the anti-TIGIT antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9; a heavy chain CDR2 sequence comprising SEQ ID The amino acid sequence of NO: 10, SEQ ID NO: 11, SEQ ID NO: 12 or SEQ ID NO: 13; and the heavy chain CDR3 sequence comprising the amine of SEQ ID NO: 14, SEQ ID NO: 15 or 16 base acid sequence.

在一些實施例中，抗TIGIT抗體包含：輕鏈CDR1序列，其包含SEQ ID NO: 17之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO: 18之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO: 19之胺基酸序列。 In some embodiments, the anti-TIGIT antibody comprises: a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 17; a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 18; and a light chain A CDR3 sequence comprising the amino acid sequence of SEQ ID NO:19.

在一些實施例中，抗TIGIT抗體包含：重鏈CDR1序列，其包含SEQ ID NO: 7、SEQ ID NO: 8或SEQ ID NO: 9之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO: 10、SEQ ID NO: 11、SEQ ID NO: 12或SEQ ID NO: 13之胺基酸序列；重鏈CDR3序列，其包含SEQ ID NO: 14、SEQ ID NO: 15或SEQ ID NO: 16之胺基酸序列；輕鏈CDR1序列，其包含SEQ ID NO: 17之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO: 18之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO: 19之胺基酸序列。In some embodiments, the anti-TIGIT antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 7, SEQ ID NO: 8 or SEQ ID NO: 9; a heavy chain CDR2 sequence comprising SEQ ID The amino acid sequence of NO: 10, SEQ ID NO: 11, SEQ ID NO: 12 or SEQ ID NO: 13; heavy chain CDR3 sequence comprising SEQ ID NO: 14, SEQ ID NO: 15 or SEQ ID NO: The amino acid sequence of 16; the light chain CDR1 sequence, which comprises the amino acid sequence of SEQ ID NO: 17; the light chain CDR2 sequence, which comprises the amino acid sequence of SEQ ID NO: 18; and the light chain CDR3 sequence, which Contains the amino acid sequence of SEQ ID NO: 19.

在一些實施例中，抗TIGIT抗體包含：包含以下之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3： (a)    分別SEQ ID NO: 7、10、14、17、18及19；或 (b)    分別SEQ ID NO: 8、11、14、17、18及19；或 (c)    分別SEQ ID NO: 9、12、15、17、18及19；或 (d)    分別SEQ ID NO: 8、13、16、17、18及19；或 (e)    分別SEQ ID NO: 8、12、16、17、18及19。 In some embodiments, the anti-TIGIT antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the following amino acid sequences: (a) SEQ ID NOs: 7, 10, 14, 17, 18 and 19, respectively; or (b) SEQ ID NOs: 8, 11, 14, 17, 18 and 19, respectively; or (c) SEQ ID NOs: 9, 12, 15, 17, 18 and 19, respectively; or (d) SEQ ID NOs: 8, 13, 16, 17, 18 and 19, respectively; or (e) SEQ ID NOs: 8, 12, 16, 17, 18 and 19, respectively.

在一些實施例中，抗TIGIT抗體包含重鏈可變區(VH)，其包含與SEQ ID NO: 1、SEQ ID NO: 2、SEQ ID NO: 3、SEQ ID NO: 4或SEQ ID NO: 5具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗TIGIT抗體包含VH，其包含SEQ ID NO: 1、SEQ ID NO: 2、SEQ ID NO: 3、SEQ ID NO: 4或SEQ ID NO: 5之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO: 1、SEQ ID NO: 2、SEQ ID NO: 3、SEQ ID NO: 4或SEQ ID NO: 5)具有至少90%序列一致性之VH序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類TIGIT且視情況仍能夠阻斷CD155及/或CD112與TIGIT之結合。In some embodiments, the anti-TIGIT antibody comprises a heavy chain variable region (VH) comprising the same sequence as SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5 have at least 90% sequence identity (e.g., at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequence. In some embodiments, the anti-TIGIT antibody comprises a VH comprising the amino acid sequence of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5. In some embodiments, a VH having at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5) the sequence contains one, two, three, four, five, six, seven, eight, nine, ten or more substitutions (eg conservative substitutions), insertions or deletions relative to the reference sequence, However, it still binds to human TIGIT and optionally blocks the binding of CD155 and/or CD112 to TIGIT.

在一些實施例中，抗TIGIT抗體包含輕鏈可變區(VL)，其包含與SEQ ID NO: 6具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗TIGIT抗體包含VL，其包含SEQ ID NO: 6之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO: 6)具有至少90%序列一致性之VL序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類TIGIT且視情況仍能夠阻斷CD155及/或CD112與TIGIT之結合。In some embodiments, the anti-TIGIT antibody comprises a light chain variable region (VL) comprising at least 90% sequence identity to SEQ ID NO: 6 (e.g., at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-TIGIT antibody comprises VL, which comprises the amino acid sequence of SEQ ID NO:6. In some embodiments, a VL sequence with at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 6) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg conservative substitutions), insertions or deletions, but still bind to human TIGIT and optionally block CD155 and/or CD112 binding to TIGIT.

在一些實施例中，抗TIGIT抗體包含重鏈可變區，其包含與SEQ ID NO: 1、SEQ ID NO: 2、SEQ ID NO: 3、SEQ ID NO: 4或SEQ ID NO: 5具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列；且包含輕鏈可變區，其包含與SEQ ID NO: 6具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%、或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗TIGIT抗體包含重鏈可變區，其包含SEQ ID NO: 1、SEQ ID NO: 2、SEQ ID NO: 3、SEQ ID NO: 4或SEQ ID NO: 5之胺基酸序列，且包含輕鏈可變區，其包含SEQ ID NO: 6之胺基酸序列。In some embodiments, the anti-TIGIT antibody comprises a heavy chain variable region comprising at least a Amino acids with 90% sequence identity (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) and comprising a light chain variable region comprising at least 90% sequence identity to SEQ ID NO: 6 (e.g. at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96% , at least 97%, at least 98%, or at least 99% sequence identity). In some embodiments, the anti-TIGIT antibody comprises a heavy chain variable region comprising the amine group of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, or SEQ ID NO: 5 acid sequence, and comprising the light chain variable region, which comprises the amino acid sequence of SEQ ID NO:6.

在一些實施例中，抗TIGIT抗體包含： (a)    包含SEQ ID NO: 1之胺基酸序列的VH及包含SEQ ID NO: 6之胺基酸序列的VL； (b)    包含SEQ ID NO: 2之胺基酸序列的VH及包含SEQ ID NO: 6之胺基酸序列的VL；或 (c)    包含SEQ ID NO: 3之胺基酸序列的VH及包含SEQ ID NO: 6之胺基酸序列的VL；或 (d)    包含SEQ ID NO: 4之胺基酸序列的VH及包含SEQ ID NO: 6之胺基酸序列的VL；或 (f)    包含SEQ ID NO: 5之胺基酸序列的VH及包含SEQ ID NO: 6之胺基酸序列的VL。 In some embodiments, the anti-TIGIT antibody comprises: (a) a VH comprising the amino acid sequence of SEQ ID NO: 1 and a VL comprising the amino acid sequence of SEQ ID NO: 6; (b) a VH comprising the amino acid sequence of SEQ ID NO: 2 and a VL comprising the amino acid sequence of SEQ ID NO: 6; or (c) a VH comprising the amino acid sequence of SEQ ID NO: 3 and a VL comprising the amino acid sequence of SEQ ID NO: 6; or (d) a VH comprising the amino acid sequence of SEQ ID NO: 4 and a VL comprising the amino acid sequence of SEQ ID NO: 6; or (f) a VH comprising the amino acid sequence of SEQ ID NO: 5 and a VL comprising the amino acid sequence of SEQ ID NO: 6.

在一些實施例中，抗TIGIT抗體包含：包含選自SEQ ID NO: 20、21、22、23及24之胺基酸序列的重鏈；及包含SEQ ID NO: 25之胺基酸序列的輕鏈。In some embodiments, the anti-TIGIT antibody comprises: a heavy chain comprising an amino acid sequence selected from the group consisting of SEQ ID NO: 20, 21, 22, 23 and 24; and a light comprising the amino acid sequence of SEQ ID NO: 25 chain.

在一些實施例中，用於本發明方法之抗TIGIT抗體為US 2009/0258013、US 2016/0176963、US 2016/0376365或WO 2016/028656中所揭示之抗TIGIT抗體的未經岩藻糖基化版本。 C. 例示性抗 CD40 抗體 In some embodiments, the anti-TIGIT antibodies used in the methods of the invention are unfucosylated anti-TIGIT antibodies disclosed in US 2009/0258013, US 2016/0176963, US 2016/0376365 or WO 2016/028656 Version. C. Exemplary Anti- CD40 Antibodies

如上文所提及，在一些實施例中，結合免疫細胞接合子之抗體為促效劑抗CD40抗體。包括達西珠單抗(dacetuzumab)之促效CD40單株抗體已在單一藥劑及組合化學療法背景下展示令人鼓舞的臨床活性。達西珠單抗在NHL之1期研究及彌漫性大B細胞淋巴瘤(DLBCL)之2期研究中展現一定臨床活性。 參加例如Advani等人， J. Clin. Oncol. 27:4371-4377 (2009)及De Vos等人， J. Hematol. Oncol. 7:1-9 (2014)。此外，當與太平洋紫杉醇或卡鉑(carboplatin)或吉西他濱(gemcitabine)組合時，針對CD40之人類化IgG2促效劑抗體CP-870,893在實體腫瘤適應症中展示令人鼓舞的活性。在此等研究中，可見抗原呈遞細胞活化、細胞介素產生及抗原特異性T細胞產生。 參見例如Beatty等人， Clin. Cancer Res. 19:6286-6295 (2013)及Vonderheide等人， Oncoimmunology2:e23033 (2013)。 As mentioned above, in some embodiments, the antibody that binds the immune cell engager is an agonist anti-CD40 antibody. Agonistic CD40 monoclonal antibodies, including dacetuzumab, have demonstrated encouraging clinical activity in both single-agent and combination chemotherapy settings. Dacilizumab has demonstrated clinical activity in a Phase 1 study in NHL and a Phase 2 study in diffuse large B-cell lymphoma (DLBCL). See eg , Advani et al, J. Clin. Oncol . 27:4371-4377 (2009) and De Vos et al, J. Hematol. Oncol . 7:1-9 (2014). In addition, the humanized IgG2 agonist antibody against CD40, CP-870,893, showed encouraging activity in solid tumor indications when combined with paclitaxel or carboplatin or gemcitabine. In these studies, antigen-presenting cell activation, interleukin production, and antigen-specific T cell production were seen. See, eg , Beatty et al., Clin. Cancer Res . 19:6286-6295 (2013) and Vonderheide et al., Oncoimmunology 2:e23033 (2013).

在一些實施例中，提供未經岩藻糖基化抗CD40抗體用於本發明方法。在一些實施例中，未經岩藻糖基化抗CD40抗體為SEA-CD40，其為鼠類S2C6之未經岩藻糖基化人類化版本且其包含分別包含SEQ ID NO: 30至35之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。對應VH及VL分別包含SEQ ID NO: 28及29之胺基酸序列。SEA-CD40描述於美國專利公開案第2017/0333556號及第2017/0137528號中，該兩件公開案以引用之方式併入本文中。S2C6最初經分離作為針對人類膀胱癌產生之鼠類單株抗體，在本文中稱作mS2C6。 參見例如Paulie等人， Cancer Immunol. Immunother. 17:165-179 (1984)。S2C6抗體為CD40傳信路徑之部分促效劑且在一些實施例中具有以下活性：結合於人類CD40蛋白、結合於食蟹獼猴CD40蛋白、活化CD40傳信路徑、增強CD40與其配位體CD40L之相互作用。 參見例如美國專利第6,946,129號。 In some embodiments, non-fucosylated anti-CD40 antibodies are provided for use in the methods of the invention. In some embodiments, the non-fucosylated anti-CD40 antibody is SEA-CD40, which is a non-fucosylated humanized version of murine S2C6 and which comprises SEQ ID NOs: 30-35, respectively Heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 of amino acid sequence. The corresponding VH and VL comprise the amino acid sequences of SEQ ID NOs: 28 and 29, respectively. SEA-CD40 is described in US Patent Publication Nos. 2017/0333556 and 2017/0137528, both of which are incorporated herein by reference. S2C6 was originally isolated as a murine monoclonal antibody raised against human bladder cancer, referred to herein as mS2C6. See, eg , Paulie et al., Cancer Immunol. Immunother . 17:165-179 (1984). The S2C6 antibody is a partial agonist of the CD40 signaling pathway and in some embodiments has the following activities: binds to human CD40 protein, binds to cynomolgus monkey CD40 protein, activates the CD40 signaling pathway, enhances the interaction between CD40 and its ligand CD40L interaction. See, eg , US Patent No. 6,946,129.

S2C6經人類化且此人類化抗體在本文中稱為人類化S2C6，且替代地稱為達西珠單抗，其為經岩藻糖基化人類化S2C6 (fhS2C6或SGN-40)。參見例如WO 2006/128103，其出於任何目的以引用的方式併入本文中。SEA-CD40為未經岩藻糖基化人類化S2C6抗體。人類化S2C6之其他版本揭示於WO2008/091954處；此等版本可未經岩藻糖基化且用於本文所揭示之方法中。S2C6 was humanized and this humanized antibody is referred to herein as humanized S2C6, and alternatively dacizumab, which is fucosylated humanized S2C6 (fhS2C6 or SGN-40). See, eg, WO 2006/128103, which is hereby incorporated by reference for any purpose. SEA-CD40 is an unfucosylated humanized S2C6 antibody. Other versions of humanized S2C6 are disclosed at WO2008/091954; such versions can be unfucosylated and used in the methods disclosed herein.

在一些實施例中，抗CD40抗體包含以下中之一或多者(例如一者、兩者、三者、四者、五者或六者)： 重鏈CDR1序列，其包含SEQ ID NO: 30之胺基酸序列； 重鏈CDR2序列，其包含SEQ ID NO: 31或SEQ ID NO: 36之胺基酸序列； 重鏈CDR3序列，其包含SEQ ID NO: 32之胺基酸序列； 輕鏈CDR1序列，其包含SEQ ID NO: 33之胺基酸序列； 輕鏈CDR2序列，其包含SEQ ID NO:34之胺基酸序列；及/或 輕鏈CDR3序列，其包含SEQ ID NO: 35之胺基酸序列。 In some embodiments, the anti-CD40 antibody comprises one or more of the following (eg, one, two, three, four, five, or six): A heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 30; A heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 31 or SEQ ID NO: 36; A heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 32; A light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 33; A light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 34; and/or A light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:35.

在一些實施例中，抗CD40抗體包含：重鏈CDR1序列，其包含SEQ ID NO: 30之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO: 31或SEQ ID NO: 36中之任一者之胺基酸序列；以及重鏈CDR3序列，其包含SEQ ID NO: 32之胺基酸序列。 In some embodiments, the anti-CD40 antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 30; a heavy chain CDR2 sequence comprising any of SEQ ID NO: 31 or SEQ ID NO: 36 The amino acid sequence of one; and the heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:32.

在一些實施例中，抗CD40抗體包含：輕鏈CDR1序列，其包含SEQ ID NO: 33之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO: 34之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO: 35之胺基酸序列。 In some embodiments, the anti-CD40 antibody comprises: a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 33; a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 34; and a light chain A CDR3 sequence comprising the amino acid sequence of SEQ ID NO:35.

在一些實施例中，抗CD40抗體包含：重鏈CDR1序列，其包含SEQ ID NO: 30之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO: 31或SEQ ID NO: 36之胺基酸序列；重鏈CDR3序列，其包含SEQ ID NO: 32之胺基酸序列；輕鏈CDR1序列，其包含SEQ ID NO: 33之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO: 34之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO: 35之胺基酸序列。In some embodiments, the anti-CD40 antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:30; a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:31 or SEQ ID NO:36 acid sequence; heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 32; light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 33; light chain CDR2 sequence comprising SEQ ID NO: The amino acid sequence of 34; and the light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO: 35.

在一些實施例中，抗CD40抗體包含：包含以下之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3： (a)    分別SEQ ID NO: 30、31、33、34及35；或 (b)    分別SEQ ID NO: 30、36、33、34及35。 In some embodiments, the anti-CD40 antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the following amino acid sequences: (a) SEQ ID NOs: 30, 31, 33, 34 and 35, respectively; or (b) SEQ ID NOs: 30, 36, 33, 34 and 35, respectively.

在一些實施例中，抗CD40抗體包含重鏈可變區(VH)，其包含與SEQ ID NO:28具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗CD40抗體包含VH，其包含NO:28之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO:28)具有至少90%序列一致性之VH序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類CD40。In some embodiments, the anti-CD40 antibody comprises a heavy chain variable region (VH) comprising at least 90% sequence identity to SEQ ID NO: 28 (eg, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-CD40 antibody comprises a VH comprising the amino acid sequence of NO:28. In some embodiments, a VH sequence with at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 28) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg, conservative substitutions), insertions or deletions, but still capable of binding to human CD40.

在一些實施例中，抗CD40抗體包含輕鏈可變區(VL)，其包含與SEQ ID NO:29具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗CD40抗體包含VL，其包含SEQ ID NO:29之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO:29)具有至少90%序列一致性之VL序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類CD40。In some embodiments, the anti-CD40 antibody comprises a light chain variable region (VL) comprising at least 90% sequence identity to SEQ ID NO:29 (eg, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-CD40 antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:29. In some embodiments, a VL sequence with at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 29) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg, conservative substitutions), insertions or deletions, but still capable of binding to human CD40.

在一些實施例中，抗CD40抗體包含重鏈可變區，其包含與SEQ ID NO:28具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列，且包含輕鏈可變區，其包含與SEQ ID NO:29具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗CD40抗體包含重鏈可變區，其包含SEQ ID NO:28之胺基酸序列，且包含輕鏈可變區，其包含SEQ ID NO:29之胺基酸序列。In some embodiments, the anti-CD40 antibody comprises a heavy chain variable region comprising at least 90% sequence identity to SEQ ID NO: 28 (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% sequence identity) amino acid sequence and comprising a light chain variable region comprising at least 90% sequence identity with SEQ ID NO:29 Amino acid sequences that are homogenous (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical in sequence identity). In some embodiments, the anti-CD40 antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:28 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:29.

在一些實施例中，抗CD40抗體包含分別揭示為SEQ ID NO:28及29之重鏈可變區及輕鏈可變區。在一些實施例中，抗CD40抗體包含分別揭示為SEQ ID NO: 26及27之重鏈及輕鏈。 D. 例示性抗 CD70 抗體 In some embodiments, the anti-CD40 antibody comprises a heavy chain variable region and a light chain variable region disclosed as SEQ ID NOs: 28 and 29, respectively. In some embodiments, the anti-CD40 antibody comprises the heavy and light chains disclosed as SEQ ID NOs: 26 and 27, respectively. D. Exemplary Anti- CD70 Antibodies

在一些實施例中，提供未經岩藻糖基化抗CD70抗體作為結合免疫細胞接合子之抗體用於本發明方法。在一些實施例中，未經岩藻糖基化抗CD70抗體為SEA-CD70，如美國專利案第8,067,546號中所描述且其包含分別包含SEQ ID NO: 53至58之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。對應VH及VL分別包含SEQ ID NO: 41及42之胺基酸序列。CD70分子為腫瘤壞死因子(TNF)配位體超家族(TNFSF)中之成員且其結合於相關受體CD27 (TNFRSF7)。兩個分子之間的相互作用活化來自兩種受體之細胞內信號。在正常條件下，CD70表現為短暫的且限於活化之T及B細胞、成熟樹突狀細胞及自然殺手(NK)細胞。類似地，CD27表現於天然且活化之效應T細胞以及NK及活化之B細胞兩者上。然而，CD70亦異常表現於各種血液癌，包括急性骨髓白血病(AML)、骨髓發育不良症候群(MDS)及非霍奇金氏淋巴瘤(NHL)以及癌瘤中，且在腫瘤細胞存活及/或腫瘤免疫逃避兩者中起作用。SEA-CD70經由阻斷CD70/CD27軸傳信、引發抗體依賴性細胞吞噬作用(ADCP)及補體依賴性細胞毒性(CDC)以及增強抗體依賴性細胞毒性(ADCC)來起作用。In some embodiments, non-fucosylated anti-CD70 antibodies are provided for use in the methods of the invention as antibodies that bind to immune cell engagers. In some embodiments, the non-fucosylated anti-CD70 antibody is SEA-CD70, as described in US Pat. No. 8,067,546 and which comprises a heavyweight comprising the amino acid sequences of SEQ ID NOs: 53-58, respectively Chains CDR1, CDR2 and CDR3 and light chains CDR1, CDR2 and CDR3. The corresponding VH and VL comprise the amino acid sequences of SEQ ID NOs: 41 and 42, respectively. The CD70 molecule is a member of the tumor necrosis factor (TNF) ligand superfamily (TNFSF) and it binds to the related receptor CD27 (TNFRSF7). The interaction between the two molecules activates intracellular signals from both receptors. Under normal conditions, CD70 is transient and restricted to activated T and B cells, mature dendritic cells and natural killer (NK) cells. Similarly, CD27 is expressed on both naive and activated effector T cells as well as NK and activated B cells. However, CD70 is also abnormally expressed in various blood cancers, including acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and non-Hodgkin's lymphoma (NHL), as well as carcinomas, and is involved in tumor cell survival and/or Tumour immune evasion plays a role in both. SEA-CD70 acts by blocking CD70/CD27 axis signaling, triggering antibody-dependent cellular phagocytosis (ADCP) and complement-dependent cytotoxicity (CDC), and enhancing antibody-dependent cellular cytotoxicity (ADCC).

在一些實施例中，抗CD70抗體包含以下中之一或多者(例如一者、兩者、三者、四者、五者或六者)： 重鏈CDR1序列，其包含SEQ ID NO:53之胺基酸序列； 重鏈CDR2序列，其包含SEQ ID NO:54之胺基酸序列； 重鏈CDR3序列，其包含SEQ ID NO:55之胺基酸序列； 輕鏈CDR1序列，其包含SEQ ID NO:56之胺基酸序列； 輕鏈CDR2序列，其包含SEQ ID NO:57之胺基酸序列；及/或 輕鏈CDR3序列，其包含SEQ ID NO:58之胺基酸序列。 In some embodiments, the anti-CD70 antibody comprises one or more of the following (eg, one, two, three, four, five, or six): A heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:53; A heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:54; A heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:55; A light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO: 56; A light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO: 57; and/or A light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:58.

在一些實施例中，抗CD70抗體包含：重鏈CDR1序列，其包含SEQ ID NO:53之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO:54之胺基酸序列；以及重鏈CDR3序列，其包含SEQ ID NO:55之胺基酸序列。 In some embodiments, the anti-CD70 antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:53; a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:54; and a heavy chain A CDR3 sequence comprising the amino acid sequence of SEQ ID NO:55.

在一些實施例中，抗CD70抗體包含：輕鏈CDR1序列，其包含SEQ ID NO:56之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO:57之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO:58之胺基酸序列。 In some embodiments, the anti-CD70 antibody comprises: a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:56; a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:57; and a light chain A CDR3 sequence comprising the amino acid sequence of SEQ ID NO:58.

在一些實施例中，抗CD70抗體包含：重鏈CDR1序列，其包含SEQ ID NO:53之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO:54之胺基酸序列；重鏈CDR3序列，其包含SEQ ID NO:55之胺基酸序列；輕鏈CDR1序列，其包含SEQ ID NO:56之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO:57之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO:58之胺基酸序列。In some embodiments, the anti-CD70 antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:53; a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:54; a heavy chain CDR3 Sequence comprising the amino acid sequence of SEQ ID NO:55; light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:56; light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:57 and the light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:58.

在一些實施例中，抗CD70抗體包含重鏈可變區(VH)，其包含與SEQ ID NO:41具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗CD70抗體包含VH，其包含NO:41之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO:41)具有至少90%序列一致性之VH序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類CD70。In some embodiments, the anti-CD70 antibody comprises a heavy chain variable region (VH) comprising at least 90% sequence identity to SEQ ID NO:41 (eg, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-CD70 antibody comprises a VH comprising the amino acid sequence of NO:41. In some embodiments, a VH sequence having at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 41 ) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg, conservative substitutions), insertions or deletions, but still capable of binding to human CD70.

在一些實施例中，抗CD70抗體包含輕鏈可變區(VL)，其包含與SEQ ID NO:42具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗CD70抗體包含VL，其包含SEQ ID NO:42之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO:42)具有至少90%序列一致性之VL序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類CD70。In some embodiments, the anti-CD70 antibody comprises a light chain variable region (VL) comprising at least 90% sequence identity to SEQ ID NO:42 (eg, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-CD70 antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:42. In some embodiments, a VL sequence with at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 42) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg, conservative substitutions), insertions or deletions, but still capable of binding to human CD70.

在一些實施例中，抗CD70抗體包含重鏈可變區，其包含與SEQ ID NO:41具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列，且包含輕鏈可變區，其包含與SEQ ID NO:42具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗CD70抗體包含重鏈可變區，其包含SEQ ID NO:41之胺基酸序列，且包含輕鏈可變區，其包含SEQ ID NO:42之胺基酸序列。In some embodiments, the anti-CD70 antibody comprises a heavy chain variable region comprising at least 90% sequence identity to SEQ ID NO:41 (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequence, and comprising a light chain variable region comprising at least 90% sequence identity with SEQ ID NO:42 Amino acid sequences that are homogenous (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical in sequence identity). In some embodiments, the anti-CD70 antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:41 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:42.

在一些實施例中，抗CD70抗體包含分別揭示為SEQ ID NO:41及42之重鏈可變區及輕鏈可變區。 E. 例示性抗 BCMA 抗體 In some embodiments, the anti-CD70 antibody comprises a heavy chain variable region and a light chain variable region disclosed as SEQ ID NOs: 41 and 42, respectively. E. Exemplary Anti- BCMA Antibodies

在一些實施例中，提供未經岩藻糖基化抗BCMA抗體作為結合免疫細胞接合子之抗體用於本發明方法。在一些實施例中，未經岩藻糖基化抗BCMA抗體為SEA-BCMA，其為靶向B細胞成熟抗原(BCMA)之抗體且其包含分別包含SEQ ID NO: 47至52之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。對應VH及VL分別包含SEQ ID NO: 45及46之胺基酸序列。BCMA表現於多發性骨髓瘤(MM)上。抗體經由阻斷配位體介導之BCMA細胞傳信、抗體依賴性細胞吞噬作用(ADCP)及增強抗體依賴性細胞細胞毒性(ADCC)來起作用。In some embodiments, non-fucosylated anti-BCMA antibodies are provided for use in the methods of the invention as antibodies that bind to immune cell engagers. In some embodiments, the unfucosylated anti-BCMA antibody is SEA-BCMA, which is an antibody targeting B cell maturation antigen (BCMA) and which comprises amino acids comprising SEQ ID NOs: 47-52, respectively Sequences of heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3. The corresponding VH and VL comprise the amino acid sequences of SEQ ID NOs: 45 and 46, respectively. BCMA is manifested in multiple myeloma (MM). Antibodies work by blocking ligand-mediated BCMA cellular signaling, antibody-dependent cellular phagocytosis (ADCP), and enhanced antibody-dependent cellular cytotoxicity (ADCC).

在一些實施例中，抗BCMA抗體包含以下中之一或多者(例如一者、兩者、三者、四者、五者或六者)： 重鏈CDR1序列，其包含SEQ ID NO:47之胺基酸序列； 重鏈CDR2序列，其包含SEQ ID NO:48之胺基酸序列； 重鏈CDR3序列，其包含SEQ ID NO:49之胺基酸序列； 輕鏈CDR1序列，其包含SEQ ID NO:50之胺基酸序列； 輕鏈CDR2序列，其包含SEQ ID NO 51之胺基酸序列；及/或 輕鏈CDR3序列，其包含SEQ ID NO:52之胺基酸序列。 In some embodiments, the anti-BCMA antibody comprises one or more of the following (eg, one, two, three, four, five, or six): A heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:47; A heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:48; A heavy chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:49; A light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:50; A light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO 51; and/or A light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:52.

在一些實施例中，抗BCMA抗體包含：重鏈CDR1序列，其包含SEQ ID NO:47之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO:48之胺基酸序列；以及重鏈CDR3序列，其包含SEQ ID NO:49之胺基酸序列。 In some embodiments, the anti-BCMA antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:47; a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:48; and a heavy chain A CDR3 sequence comprising the amino acid sequence of SEQ ID NO:49.

在一些實施例中，抗BCMA抗體包含：輕鏈CDR1序列，其包含SEQ ID NO:50之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO:51之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO:52之胺基酸序列。 In some embodiments, the anti-BCMA antibody comprises: a light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:50; a light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:51; and a light chain A CDR3 sequence comprising the amino acid sequence of SEQ ID NO:52.

在一些實施例中，抗BCMA抗體包含：重鏈CDR1序列，其包含SEQ ID NO:47之胺基酸序列；重鏈CDR2序列，其包含SEQ ID NO:48之胺基酸序列；重鏈CDR3序列，其包含SEQ ID NO:49之胺基酸序列；輕鏈CDR1序列，其包含SEQ ID NO:50之胺基酸序列；輕鏈CDR2序列，其包含SEQ ID NO:51之胺基酸序列；以及輕鏈CDR3序列，其包含SEQ ID NO:52之胺基酸序列。In some embodiments, the anti-BCMA antibody comprises: a heavy chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:47; a heavy chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:48; a heavy chain CDR3 Sequence comprising the amino acid sequence of SEQ ID NO:49; light chain CDR1 sequence comprising the amino acid sequence of SEQ ID NO:50; light chain CDR2 sequence comprising the amino acid sequence of SEQ ID NO:51 and a light chain CDR3 sequence comprising the amino acid sequence of SEQ ID NO:52.

在一些實施例中，抗BCMA抗體包含重鏈可變區(VH)，其包含與SEQ ID NO:45具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗BCMA抗體包含VH，其包含NO:45之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO:45)具有至少90%序列一致性之VH序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類BCMA。In some embodiments, the anti-BCMA antibody comprises a heavy chain variable region (VH) comprising at least 90% sequence identity to SEQ ID NO:45 (eg, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-BCMA antibody comprises a VH comprising the amino acid sequence of NO:45. In some embodiments, a VH sequence having at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 45) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg, conservative substitutions), insertions or deletions, but still capable of binding to human BCMA.

在一些實施例中，抗BCMA抗體包含輕鏈可變區(VL)，其包含與SEQ ID NO:46具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗BCMA抗體包含VL，其包含SEQ ID NO:46之胺基酸序列。在一些實施例中，與參考序列(例如SEQ ID NO:46)具有至少90%序列一致性之VL序列相對於參考序列含有一個、兩個、三個、四個、五個、六個、七個、八個、九個、十個或更多個取代(例如保守性取代)、插入或缺失，但仍能夠結合於人類BCMA。In some embodiments, the anti-BCMA antibody comprises a light chain variable region (VL) comprising at least 90% sequence identity to SEQ ID NO:46 (eg, at least 91%, at least 92%, at least 93%, at least 94%) %, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequences. In some embodiments, the anti-BCMA antibody comprises a VL comprising the amino acid sequence of SEQ ID NO:46. In some embodiments, a VL sequence with at least 90% sequence identity to a reference sequence (eg, SEQ ID NO: 46) contains one, two, three, four, five, six, seven relative to the reference sequence One, eight, nine, ten or more substitutions (eg, conservative substitutions), insertions or deletions, but still capable of binding to human BCMA.

在一些實施例中，抗BCMA抗體包含重鏈可變區，其包含與SEQ ID NO:45具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列，且包含輕鏈可變區，其包含與SEQ ID NO:46具有至少90%序列一致性(例如至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列。在一些實施例中，抗BCMA抗體包含重鏈可變區，其包含SEQ ID NO:45之胺基酸序列，且包含輕鏈可變區，其包含SEQ ID NO:46之胺基酸序列。In some embodiments, the anti-BCMA antibody comprises a heavy chain variable region comprising at least 90% sequence identity to SEQ ID NO:45 (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) amino acid sequence, and comprising a light chain variable region comprising at least 90% sequence identity with SEQ ID NO:46 Amino acid sequences that are homogenous (eg, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical in sequence identity). In some embodiments, the anti-BCMA antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:45 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:46.

在一些實施例中，抗BCMA抗體包含分別揭示為SEQ ID NO:45及46之重鏈可變區及輕鏈可變區。 F. 增強型Fc主鏈 In some embodiments, the anti-BCMA antibody comprises a heavy chain variable region and a light chain variable region disclosed as SEQ ID NOs: 45 and 46, respectively. F. Enhanced Fc Backbone

如上文所提及，結合免疫細胞接合子之抗體包含具有以下活性中之一或多者的Fc：增強與一或多種活化FcγR之結合；減弱與抑制性FcγR之結合；增強ADCC活性及/或增強ADCP活性。具有帶此類活性及所需活性概況之Fc的抗體可以多種方式產生，包括產生未經岩藻糖基化蛋白及/或藉由將Fc工程化以含有產生所需活性之某些突變。此部分提供關於產生未經岩藻糖基化抗體及例示性工程化途徑之方法的額外細節。關於恆定區之選擇及抗體之製造的額外指導提供於下文其他部分中。As mentioned above, an antibody that binds an immune cell engager comprises an Fc having one or more of the following activities: enhance binding to one or more activating FcγRs; reduce binding to inhibitory FcγRs; enhance ADCC activity and/or Enhance ADCP activity. Antibodies with Fcs with such activities and desired activity profiles can be produced in a variety of ways, including by producing unfucosylated proteins and/or by engineering the Fc to contain certain mutations that produce the desired activity. This section provides additional details on methods for producing non-fucosylated antibodies and exemplary engineered pathways. Additional guidance on selection of constant regions and manufacture of antibodies is provided elsewhere below.

抗體可在其恆定區中之保守位置處經糖基化(Jefferis及Lund, (1997) Chem. Immunol. 65:111-128；Wright及Morrison, (1997) TibTECH 15:26-32)。免疫球蛋白之寡糖側鏈影響蛋白質之功能(Boyd等人, (1996) Mol. Immunol. 32:1311-1318；Wittwe及Howard, (1990) Biochem. 29:4175-4180)及糖蛋白之部分之間的分子內相互作用，該相互作用可能影響糖蛋白之構形及所呈現之三維表面(Jefferis及Lund，見上文；Wyss及Wagner, (1996) Current Op. Biotech. 7:409-416)。寡糖亦可用於基於特定識別結構而將給定糖蛋白靶向至某些分子。舉例而言，已報導在無乳糖化(agalactosylated) IgG中，CH2間空間及末端N-乙醯基葡糖胺殘基外『翻轉』出之寡糖部分變成可用以結合甘露糖結合蛋白(Malhotra等人, (1995) Nature Med. 1:237-243)。自中國倉鼠卵巢(CHO)細胞中產生之CAMPATH-1H (識別人類淋巴球之CDw52抗原的重組人類化鼠類單株IgG1抗體)移除寡糖之醣肽導致補體介導之裂解(CMCL)完全減小(Boyd 等人 ,(1996) Mol. Immunol. 32:1311-1318)，而使用神經胺糖酸酶選擇性移除唾液酸殘基未導致DMCL損失。亦報導抗體之糖基化影響抗體依賴性細胞毒性(ADCC)。特定言之，據報導，具有β(1,4)-N-乙醯胺基葡萄糖轉移酶III (GnTIII) (一種催化二等分GlcNAc之形成之糖基轉移酶)之四環素調節表現之CHO細胞具有提高的ADCC活性(Umana等人，(1999) Mature Biotech. 17:176-180)。 Antibodies can be glycosylated at conserved positions in their constant regions (Jefferis and Lund, (1997) Chem. Immunol. 65:111-128; Wright and Morrison, (1997) TibTECH 15:26-32). Oligosaccharide side chains of immunoglobulins affect protein function (Boyd et al., (1996) Mol. Immunol. 32:1311-1318; Wittwe and Howard, (1990) Biochem. 29:4175-4180) and portions of glycoproteins Intramolecular interactions between glycoproteins that may affect the conformation of glycoproteins and the presented three-dimensional surface (Jefferis and Lund, supra; Wyss and Wagner, (1996) Current Op. Biotech. 7:409-416 ). Oligosaccharides can also be used to target a given glycoprotein to certain molecules based on specific recognition structures. For example, it has been reported that in agalactosylated IgG, the oligosaccharide moiety "flipped" out of the inter-CH2 space and the terminal N-acetylglucosamine residue becomes available for binding to mannose-binding protein (Malhotra et al, (1995) Nature Med. 1:237-243). Removal of oligosaccharide glycopeptides from CAMPATH-1H (a recombinant humanized murine monoclonal IgG1 antibody recognizing the CDw52 antigen of human lymphocytes) produced in Chinese hamster ovary (CHO) cells results in complete complement-mediated lysis (CMCL) reduced (Boyd et al ., (1996) Mol. Immunol. 32:1311-1318), while selective removal of sialic acid residues using neuraminidase did not result in DMCL loss. Glycosylation of antibodies has also been reported to affect antibody-dependent cellular cytotoxicity (ADCC). Specifically, CHO cells were reported to have tetracycline-regulated performance of β(1,4)-N-acetylaminoglucosyltransferase III (GnTIII), a glycosyltransferase that catalyzes the formation of bisecting GlcNAc Has increased ADCC activity (Umana et al. (1999) Mature Biotech. 17:176-180).

抗體之糖基化通常為N鍵聯或O鍵聯。N鍵聯係指碳水化合物部分與天冬醯胺殘基之側鏈連接。三肽序列天冬醯胺-X-絲胺酸及天冬醯胺-X-蘇胺酸，其中X為除脯胺酸以外之任何胺基酸，為用於碳水化合物部分與天冬醯胺側鏈之酶促連接之識別序列。因此，多肽中存在此等三肽序列中之任一者會產生潛在糖基化位點。O鍵聯之糖基化係指糖N-乙醯半乳胺糖、半乳糖或木糖中之一者與羥胺基酸，最通常絲胺酸或蘇胺酸之連接，但亦可使用5-羥基脯胺酸或5-羥基離胺酸。Glycosylation of antibodies is usually N-linked or O-linked. N linkage refers to the attachment of the carbohydrate moiety to the side chain of the asparagine residue. The tripeptide sequences asparagine-X-serine and asparagine-X-threonine, where X is any amino acid except proline, are used for the carbohydrate moiety and asparagine Recognition sequences for enzymatic ligation of side chains. Thus, the presence of any of these tripeptide sequences in a polypeptide creates a potential glycosylation site. O-linked glycosylation refers to the attachment of one of the sugars N-acetylgalactosamine, galactose or xylose to a hydroxylamine acid, most commonly serine or threonine, but 5 can also be used -Hydroxyproline or 5-hydroxylysine.

抗體之糖基化變體為其中抗體之糖基化模式經改變之變異體。改變意謂刪除抗體中發現的一或多個碳水化合物部分，將一或多個碳水化合物部分添加至抗體，改變糖基化組成(糖基化模式)、糖基化程度等。A glycosylation variant of an antibody is one in which the glycosylation pattern of the antibody is altered. Altering means deleting one or more carbohydrate moieties found in the antibody, adding one or more carbohydrate moieties to the antibody, altering the glycosylation composition (glycosylation pattern), degree of glycosylation, and the like.

將糖基化位點添加至抗體可藉由改變胺基酸序列，使得其含有上述三肽序列(用於N鍵聯糖基化位點)中之一或多者來實現。亦可藉由向原始抗體之序列添加一或多個絲胺酸或蘇胺酸殘基或用該等殘基取代原始抗體之序列來進行改變(用於O鍵聯的糖基化位點)。類似地，移除糖基化位點可藉由抗體之天然糖基化位點內之胺基酸改變來實現。Addition of glycosylation sites to the antibody can be accomplished by altering the amino acid sequence such that it contains one or more of the tripeptide sequences described above (for N-linked glycosylation sites). Changes can also be made by adding or substituting one or more serine or threonine residues to the sequence of the original antibody (for O-linked glycosylation sites) . Similarly, removal of glycosylation sites can be accomplished by amino acid changes within the antibody's native glycosylation sites.

胺基酸序列通常藉由改變下伏核酸序列而改變。此等方法包括自天然來源分離(在天然存在之胺基酸序列變體的情況下)或藉由對抗體的早期製備變異體或非變異體版本進行寡核苷酸介導(或定點)突變誘發、PCR突變誘發及卡匣突變誘發來製備。The amino acid sequence is usually altered by altering the underlying nucleic acid sequence. Such methods include isolation from natural sources (in the case of naturally-occurring amino acid sequence variants) or by oligonucleotide-mediated (or site-directed) mutagenesis of earlier prepared variant or non-variant versions of the antibody Induction, PCR mutagenesis, and cassette mutagenesis were prepared.

抗體之糖基化(包括糖基化模式)亦可在不改變胺基酸序列或下伏核苷酸序列的情況下改變。 參見例如Pereira等人，2018, MAbs, 10(5): 693-711。糖基化在很大程度上視用於表現抗體之宿主細胞而定。由於用於將重組糖蛋白，例如抗體表現為潛在治療劑之細胞類型為罕見天然細胞，因此可預期抗體之糖基化模式的明顯變化。參見例如Hse 等人，(1997) J. Biol. Chem. 272:9062-9070。除選擇宿主細胞之外，在重組產生抗體期間影響糖基化的因素包括生長模式、培養基調配、培養物密度、加氧作用、pH、純化方案及其類似因素。已提議改變特定宿主生物體中達成之糖基化模式的各種方法，包括引入或過度表現參與寡醣產生之某些酶(美國專利第5047335號；第5510261號；第5278299號)。可以酶方式(例如使用內切糖苷酶H (Endo H))自糖蛋白移除糖基化或某些類型之糖基化。另外，重組宿主細胞可經基因工程改造以例如在處理某些類型之多糖中形成缺陷。此等及類似技術為此項技術中所熟知。 Glycosylation, including glycosylation patterns, of antibodies can also be altered without altering the amino acid sequence or underlying nucleotide sequence. See, eg , Pereira et al., 2018, MAbs , 10(5): 693-711. Glycosylation depends to a large extent on the host cell used to express the antibody. Since the cell types used to express recombinant glycoproteins, such as antibodies, as potential therapeutics are rare natural cells, significant changes in the glycosylation pattern of antibodies can be expected. See, eg, Hse et al., (1997) J. Biol. Chem. 272:9062-9070. In addition to selection of host cells, factors affecting glycosylation during recombinant antibody production include growth mode, media formulation, culture density, oxygenation, pH, purification protocols, and the like. Various approaches have been proposed to alter the glycosylation pattern achieved in a particular host organism, including the introduction or overexpression of certain enzymes involved in oligosaccharide production (US Pat. Nos. 5,047,335; 5,510,261; 5,278,299). Glycosylation, or certain types of glycosylation, can be removed from glycoproteins enzymatically (eg, using Endoglycosidase H (Endo H)). In addition, recombinant host cells can be genetically engineered to develop defects, eg, in the processing of certain types of polysaccharides. These and similar techniques are well known in the art.

抗體之糖基化結構可藉由碳水化合物分析之習知技術，包括凝集素層析、NMR、質譜分析、HPLC、GPC、單糖組成分析、依序酶促消化及HPAEC-PAD容易地分析，該碳水化合物分析使用高pH陰離子交換層析以基於電荷分離寡糖。出於分析型目的用於釋放寡糖之方法亦為已知的，且包括但不限於酶促處理(通常使用肽-N-糖苷酶F/內-β-半乳糖執行)，使用刺激性鹼性環境進行消除以釋放主要O鍵聯之結構及使用無水肼釋放N及O鍵聯之寡糖兩者之化學方法。The glycosylation structure of an antibody can be readily analyzed by conventional techniques of carbohydrate analysis, including lectin chromatography, NMR, mass spectrometry, HPLC, GPC, monosaccharide composition analysis, sequential enzymatic digestion and HPAEC-PAD, This carbohydrate analysis uses high pH anion exchange chromatography to separate oligosaccharides based on charge. Methods for the release of oligosaccharides for analytical purposes are also known and include, but are not limited to, enzymatic treatment (usually performed using peptide-N-glycosidase F/endo-beta-galactose), using stimulating bases The sexual environment is eliminated to release the predominantly O-linked structure and the chemistry using anhydrous hydrazine to release both N- and O-linked oligosaccharides.

抗體之糖基化修飾之較佳形式為減少核心岩藻糖基化。「核心岩藻糖基化」係指將岩藻糖(「岩藻糖基化」)添加至N鍵聯之聚糖之還原末端處的N-乙醯基葡糖胺(「GlcNAc」)。A preferred form of glycosylation modification of the antibody is to reduce core fucosylation. "Core fucosylation" refers to the addition of fucose ("fucosylation") to N-acetylglucosamine ("GlcNAc") at the reducing end of an N-linked glycan.

「複合N-糖苷鍵聯之糖鏈」通常結合於天冬醯胺297 (根據Kabat之編號)。如本文所用，複合N-糖苷鍵聯之糖鏈具有二觸角複合糖鏈，其主要具有以下結構：

Figure 02_image001
其中 +指示糖分子可存在或不存在，且數字指示糖分子之間的鍵位置。在上文結構中，結合至天冬醯胺之糖鏈端稱為還原末端(右側)，而相對側稱為非還原末端。岩藻糖通常藉由α1,6鍵(GlcNAc之6位鍵聯至岩藻糖之1位)通常結合於還原末端之N-乙醯基葡糖胺(「GlcNAc」)。「Gal」係指半乳糖，且「Man」係指甘露糖。 A "complex N-glycosidically linked sugar chain" is usually bound to asparagine 297 (numbering according to Kabat). As used herein, a complex N-glycosidically linked sugar chain has a diantennary complex sugar chain, which mainly has the following structure:
Figure 02_image001
Where + indicates that the sugar molecule may or may not be present, and the number indicates the bond position between the sugar molecules. In the above structure, the end of the sugar chain bound to asparagine is called the reducing end (right side), and the opposite side is called the non-reducing end. Fucose is usually bound to N-acetylglucosamine ("GlcNAc") at the reducing end, typically via an alpha 1,6 bond (the 6-position of GlcNAc is bound to the 1-position of fucose). "Gal" refers to galactose, and "Man" refers to mannose.

「複合N-糖苷鍵聯之糖鏈」包括1)複合型，其中核心結構之非還原末端側具有半乳糖-N-乙醯基葡糖胺(亦稱為「gal-GlcNAc」)之零個、一或多個分支且gal-GlcNAc之非還原末端側視情況具有唾液酸、等分N-乙醯基葡糖胺或其類似物；以及2)混合型，其中核心結構之非還原末端側具有高甘露糖N-糖苷鍵聯之糖鏈及複合N-糖苷鍵聯之糖鏈之兩個分支。The "complex N-glycosidically linked sugar chain" includes 1) a complex type in which the non-reducing end side of the core structure has zero galactose-N-acetylglucosamine (also known as "gal-GlcNAc") , one or more branches and the non-reducing end of gal-GlcNAc optionally flanks sialic acid, an aliquot of N-acetylglucosamine, or an analog thereof; and 2) a mixed type in which the non-reducing end of the core structure flanks Two branches of sugar chains with high mannose N-glycosidic linkages and complex N-glycosidic linkages.

在如本文所提供之一些方法中，僅較小量之岩藻糖經併入至抗體之複合N-糖苷鍵聯之糖鏈中。舉例而言，在各種實施例中，組合物中小於約60%、小於約50%、小於約40%、小於約30%、小於約20%、小於約15%、小於約10%、小於約5%或小於約3%之抗體具有藉由岩藻糖之核心岩藻糖基化。在一些實施例中，組合物中約2%之抗體具有藉由岩藻糖之核心岩藻糖基化。在各種實施例中，當組合物中小於60%之抗體具有藉由岩藻糖之核心岩藻糖基化時，該組合物之抗體可稱為「未經岩藻糖基化」或「去岩藻糖基化」。在一些實施例中，組合物中至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%之抗體未經岩藻糖基化。In some methods as provided herein, only minor amounts of fucose are incorporated into the complex N-glycosidically linked sugar chains of the antibody. For example, in various embodiments, less than about 60%, less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5% or less than about 3% of the antibodies had core fucosylation by fucose. In some embodiments, about 2% of the antibodies in the composition have core fucosylation by fucose. In various embodiments, antibodies of the composition may be referred to as "unfucosylated" or "de-fucosylated" when less than 60% of the antibodies in the composition have core fucosylation by fucose Fucosylation". In some embodiments, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the antibodies in the composition Not fucosylated.

在某些實施例中，僅較小量之岩藻糖類似物(或岩藻糖類似物之代謝物或產物)經併入至複合N-糖苷鍵聯之糖鏈中。舉例而言，在各種實施例中，小於約60%、小於約50%、小於約40%、小於約30%、小於約20%、小於約15%、小於約10%、小於約5%或小於約3%之抗體具有藉由岩藻糖類似物或岩藻糖類似物之代謝物或產物之核心岩藻糖基化。在一些實施例中，約2%之抗體具有藉由岩藻糖類似物或岩藻糖類似物之代謝物或產物之核心岩藻糖基化。In certain embodiments, only minor amounts of the fucose analog (or a metabolite or product of the fucose analog) are incorporated into complex N-glycosidically linked sugar chains. For example, in various embodiments, less than about 60%, less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, or Less than about 3% of the antibodies had core fucosylation by a fucose analog or a metabolite or product of a fucose analog. In some embodiments, about 2% of the antibodies have core fucosylation by a fucose analog or a metabolite or product of a fucose analog.

在一些實施例中，組合物中小於約60%、小於約50%、小於約40%、小於約30%、小於約20%、小於約15%、小於約10%、小於約5%或小於約3%之抗體在G0、G1或G2聚糖結構上具有岩藻糖殘基。( 參見例如Raju等人，2012, MAbs 4: 385-391，圖3。)在一些實施例中，組合物中約2%之抗體在G0、G1或G2聚糖結構上具有岩藻糖殘基。在各種實施例中，當組合物中小於60%之抗體在G0、G1或G2聚糖結構上具有岩藻糖殘基時，該組合物之抗體可稱為「未經岩藻糖基化」。在一些實施例中，組合物中至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%之抗體在G0、G1或G2聚糖結構上缺失岩藻糖。應注意，G0聚糖包括G0-GN聚糖。G0-GN聚糖為具有一個封端GlcNAc殘基之單觸角聚糖。G1聚糖包括G1-GN聚糖。G1-GN聚糖為具有一個封端半乳糖殘基之單觸角聚糖。G0-GN及G1-GN聚糖可經岩藻糖基化或未經岩藻糖基化。 In some embodiments, less than about 60%, less than about 50%, less than about 40%, less than about 30%, less than about 20%, less than about 15%, less than about 10%, less than about 5%, or less than about 10% in the composition About 3% of the antibodies had fucose residues on the GO, G1 or G2 glycan structure. ( See eg , Raju et al., 2012, MAbs 4: 385-391, Figure 3.) In some embodiments, about 2% of the antibodies in the composition have fucose residues on the GO, G1 or G2 glycan structure . In various embodiments, antibodies of the composition may be referred to as "unfucosylated" when less than 60% of the antibodies in the composition have fucose residues on the GO, G1 or G2 glycan structure . In some embodiments, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the antibodies in the composition Fucose was deleted on the G0, G1 or G2 glycan structure. It should be noted that GO glycans include GO-GN glycans. GO-GN glycans are monoantennary glycans with one capping GlcNAc residue. G1 glycans include G1-GN glycans. G1-GN glycans are monoantennary glycans with one end-capped galactose residue. GO-GN and G1-GN glycans may or may not be fucosylated.

可採用產生未經岩藻糖基化抗體的多種方法。例示性策略包括使用缺乏參與岩藻糖基化路徑之某些生物合成酶的細胞株或抑制或敲除參與岩藻糖基化路徑之某些基因。此類途徑之綜述藉由Pereira,等人(2018) MABS 10:693-711提供，其以全文引用的方式併入本文中。Various methods of producing non-fucosylated antibodies are available. Exemplary strategies include the use of cell lines lacking certain biosynthetic enzymes involved in the fucosylation pathway or the inhibition or knockout of certain genes involved in the fucosylation pathway. A review of such approaches is provided by Pereira, et al. (2018) MABS 10:693-711, which is incorporated herein by reference in its entirety.

舉例而言，藉由將抗體生產細胞與岩藻糖類似物一起培育來製備非岩藻糖基化抗體之方法描述於例如WO2009/135181中。簡言之，在岩藻糖類似物或岩藻糖類似物之細胞內代謝物或產物之存在下培育已經工程改造以表現抗體之細胞。細胞內代謝物可為例如經GDP修飾之類似物或完全或部分去酯化類似物。舉例而言，產物可為完全或部分去酯化類似物。在一些實施例中，岩藻糖類似物可抑制岩藻糖補救路徑中之酶。舉例而言，岩藻糖類似物(或岩藻糖類似物之細胞內代謝物或產物)可抑制岩藻糖激酶或GDP-岩藻糖-焦磷酸化酶之活性。在一些實施例中，岩藻糖模擬物(或岩藻糖類似物之細胞內代謝物或產物)抑制岩藻糖基轉移酶(較佳地1,6-岩藻糖基轉移酶，例如FUT8蛋白)。在一些實施例中，岩藻糖類似物(或岩藻糖類似物之細胞內代謝物或產物)可抑制岩藻糖之重新合成路徑中之酶的活性。舉例而言，岩藻糖類似物(或岩藻糖類似物之細胞內代謝物或產物)可抑制GDP-甘露糖4,6-去水酶或/或GDP-岩藻糖合成酶之活性。在一些實施例中，岩藻糖類似物(或岩藻糖類似物之細胞內代謝物或產物)可抑制岩藻糖轉運子(例如GDP-岩藻糖轉運子)。For example, methods of making afucosylated antibodies by incubating antibody-producing cells with fucose analogs are described, for example, in WO2009/135181. Briefly, cells that have been engineered to express antibodies are grown in the presence of a fucose analog or an intracellular metabolite or product of a fucose analog. Intracellular metabolites can be, for example, GDP-modified analogs or fully or partially de-esterified analogs. For example, the product can be a fully or partially deesterified analog. In some embodiments, the fucose analogs inhibit enzymes in the fucose salvage pathway. For example, a fucose analog (or an intracellular metabolite or product of a fucose analog) can inhibit the activity of fucose kinase or GDP-fucose-pyrophosphorylase. In some embodiments, the fucose mimetic (or intracellular metabolite or product of the fucose analog) inhibits a fucosyltransferase (preferably a 1,6-fucosyltransferase, such as FUT8 protein). In some embodiments, the fucose analog (or intracellular metabolite or product of the fucose analog) inhibits the activity of enzymes in the de novo fucose synthesis pathway. For example, a fucose analog (or an intracellular metabolite or product of a fucose analog) can inhibit the activity of GDP-mannose 4,6-dehydratase or/or GDP-fucose synthase. In some embodiments, the fucose analog (or intracellular metabolite or product of the fucose analog) inhibits a fucose transporter (eg, the GDP-fucose transporter).

在一個實施例中，岩藻糖類似物為2-氟岩藻糖。在生長培養基中使用岩藻糖類似物及其他岩藻糖類似物之方法揭示於例如WO 2009/135181中，其以引用之方式併入本文中。In one embodiment, the fucose analog is 2-fluorofucose. Methods of using fucose analogs and other fucose analogs in growth media are disclosed, for example, in WO 2009/135181, which is incorporated herein by reference.

用於工程改造細胞株以減少核心岩藻糖基化的其他方法包括基因剔除、基因敲入及RNA干擾(RNAi)。 參見例如Pereira等人，2018, MAbs, 10(5): 693-711。在基因剔除中，編碼FUT8 (α1,6-岩藻糖基轉移酶)之基因不活化。FUT8催化岩藻糖基殘基自GDP-岩藻糖轉移至N-聚糖之Asn鍵聯(N鍵聯)之GlcNac的位置6。FUT8經報導為負責將岩藻糖添加至Asn297處N鍵聯之二觸角碳水化合物的唯一酶。基因敲入添加編碼諸如GNTIII或Golgi α甘露糖苷酶II之酶之基因。細胞中此類酶含量之增大使自岩藻糖基化路徑轉向單株抗體(導致核心岩藻糖基化減少)，且具有增大量的二等分N-乙醯葡萄糖胺。RNAi通常亦靶向FUT8基因表現，其導致mRNA轉錄量減小或完全地剔除基因表現。 Other methods for engineering cell lines to reduce core fucosylation include gene knockout, gene knock-in, and RNA interference (RNAi). See, eg , Pereira et al., 2018, MAbs , 10(5): 693-711. In gene knockout, the gene encoding FUT8 (α1,6-fucosyltransferase) was not activated. FUT8 catalyzes the transfer of a fucosyl residue from GDP-fucose to position 6 of the GlcNac of the Asn-linked (N-linked) N-glycan. FUT8 was reported to be the only enzyme responsible for the addition of fucose to the N-linked diantennary carbohydrate at Asn297. Knock-in adds genes encoding enzymes such as GNTIII or Golgi alpha mannosidase II. Increased levels of these enzymes in cells diverted from the fucosylation pathway to monoclonal antibodies (resulting in reduced core fucosylation) with increased amounts of bisected N-acetylglucosamine. RNAi also typically targets the FUT8 gene expression, which results in reduced mRNA transcript levels or complete deletion of the gene expression.

可使用之其他策略包括GlycoMAb ®(美國專利案第6,602,684號)及Potelligent ®(BioWa)。 Other strategies that can be used include GlycoMAb ® (US Patent No. 6,602,684) and Potelligent ® (BioWa).

此等方法中之任一者可用於產生將能夠產生未經岩藻糖基化抗體的細胞株。Any of these methods can be used to generate cell lines that will be capable of producing non-fucosylated antibodies.

各種工程化途徑亦可用以獲得具有所需FcγR活性及效應功能之Fc區。在一些實施例中，Fc經工程化以具有以下突變組合：S239D、A330L及I332E，其增大Fc域對FcγRIIIA之親和力且因此增大ADCC。增強對FcγRIIIa之親和力的額外取代包括例如T256A、K290A、S298A、E333A及K334A。增強與活化FcγRIIIa之結合且減弱與抑制性FcγRIIIb之結合的取代包括例如F243L/R292P/Y300L/V305I/P396L及F243L/R292P/Y300L/L235V/P396L。在一些實施例中，取代處於IgG1 Fc背景下。Various engineering approaches can also be used to obtain Fc regions with desired FcyR activity and effector function. In some embodiments, the Fc is engineered with the following combination of mutations: S239D, A330L, and I332E, which increase the affinity of the Fc domain for FcyRIIIA and thus increase ADCC. Additional substitutions that enhance affinity for FcyRIIIa include, for example, T256A, K290A, S298A, E333A, and K334A. Substitutions that enhance binding to activating FcγRIIIa and decrease binding to inhibitory FcγRIIIb include, for example, F243L/R292P/Y300L/V305I/P396L and F243L/R292P/Y300L/L235V/P396L. In some embodiments, the substitution is in the context of an IgGl Fc.

共價連接至保守性Asn297之寡糖涉及IgG之Fc區結合FcγR之能力(Lund 等人, 1996, J. Immunol.157:4963-69；Wright及Morrison, 1997, Trends Biotechnol.15:26-31)。IgG上此糖型之工程化可顯著提高IgG介導之ADCC。將二等分N-乙醯基葡糖胺修飾(Umana 等人, 1999, Nat. Biotechnol.17:176-180；Davies 等人, 2001, Biotech. Bioeng.74:288-94)添加至此糖型或自此糖型移除岩藻糖(Shields等人, 2002, J. Biol. Chem.277:26733-40；Shinkawa等人, 2003, J. Biol. Chem.278:6591-604；Niwa等人, 2004, Cancer Res.64:2127-33)為提高IgG Fc與FcγR之間的結合，由此增強Ig介導之ADCC活性的IgG Fc工程化之兩個實例。 Oligosaccharides covalently linked to the conserved Asn297 are involved in the ability of the Fc region of IgG to bind FcγRs (Lund et al ., 1996, J. Immunol. 157:4963-69; Wright and Morrison, 1997, Trends Biotechnol. 15:26-31 ). Engineering of this glycoform on IgG can significantly enhance IgG-mediated ADCC. A bisected N-acetylglucosamine modification (Umana et al ., 1999, Nat. Biotechnol. 17:176-180; Davies et al ., 2001, Biotech. Bioeng. 74:288-94) was added to this glycoform or removal of fucose from this glycoform (Shields et al., 2002, J. Biol. Chem. 277:26733-40; Shinkawa et al., 2003, J. Biol. Chem. 278:6591-604; Niwa et al. , 2004, Cancer Res. 64:2127-33) are two examples of IgG Fc engineering to increase binding between IgG Fc and FcyRs, thereby enhancing Ig-mediated ADCC activity.

人類IgG1 Fc區之溶劑暴露胺基酸之全身性取代已產生具有改變的FcγR結合親和力的IgG變異體(Shields 等人，2001, J. Biol. Chem.276:6591-604)。當與親本IgG1相比時，涉及Thr256/Ser298、Ser298/Glu333、Ser298/Lys334或Ser298/Glu333/Lys334處取代Ala之此等變異體之子集展現針對FcγR之結合親和力及ADCC活性兩者均增大(Shields 等人 ，2001, J. Biol. Chem.276:6591-604；Okazaki 等人，2004, J. Mol. Biol.336:1239-49)。 Systemic substitution of solvent-exposed amino acids of the human IgGl Fc region has produced IgG variants with altered FcγR binding affinity (Shields et al ., 2001, J. Biol. Chem. 276:6591-604). A subset of these variants involving substitutions for Ala at Thr256/Ser298, Ser298/Glu333, Ser298/Lys334 or Ser298/Glu333/Lys334 exhibited both increased binding affinity and ADCC activity for FcγRs when compared to the parental IgG1 Large (Shields et al ., 2001, J. Biol. Chem. 276:6591-604; Okazaki et al., 2004, J. Mol. Biol. 336:1239-49).

多種方法可用以測定抗體上之岩藻糖基化的量。方法包括例如經由PLRP-S層析之LC-MS、電噴霧電離四極TOF MS、毛細管電泳與雷射誘導之螢光法( CE−LIF)及親水性相互作用層析與螢光偵測(HILIC)。 IV. 例示性抗體-藥物結合物(ADC) Various methods can be used to determine the amount of fucosylation on an antibody. Methods include, for example, LC-MS via PLRP-S chromatography, electrospray ionization quadrupole TOF MS, capillary electrophoresis and laser-induced fluorescence ( CE-LIF ), and hydrophilic interaction chromatography with fluorescence detection (HILIC). ). IV. Exemplary Antibody-Drug Conjugates (ADCs)

先前部分描述了結合於參與免疫調節之目標(免疫細胞接合子)之抗體之相關態樣。如上文所提及，本文中所提供之一些方法亦包含投與包含微管蛋白干擾劑(例如奧瑞他汀，包括例如MMAE及MMAF)與結合於免疫細胞接合子之抗體的組合之抗體-藥物結合物(ADC)。在各種實施例中，抗體-藥物結合物包含與細胞毒性劑結合之抗體。在一些實施例中，細胞毒性劑為微管蛋白干擾劑。在一些實施例中，抗體結合於表現於腫瘤細胞上之抗原。關於用於本文所提供之方法中之ADC的其他細節闡述於此部分及下方實例中。本文所描述之任何ADC可與結合本文所描述之免疫細胞接合子的任何抗體組合。 A. 例示性目標抗原 The previous section describes related aspects of antibodies that bind to targets involved in immune regulation (immune cell engagers). As mentioned above, some of the methods provided herein also include administering an antibody-drug comprising a combination of a tubulin interfering agent, such as auristatin, including, for example, MMAE and MMAF, and an antibody that binds to an immune cell engager Conjugates (ADCs). In various embodiments, the antibody-drug conjugate comprises an antibody conjugated to a cytotoxic agent. In some embodiments, the cytotoxic agent is a tubulin interfering agent. In some embodiments, the antibody binds to an antigen expressed on tumor cells. Additional details regarding ADCs used in the methods provided herein are set forth in this section and in the Examples below. Any of the ADCs described herein can be combined with any antibody that binds the immune cell engagers described herein. A. Exemplary Target Antigens

在一些實施例中，ADC結合表現於腫瘤細胞上之抗原。In some embodiments, the ADC binds to an antigen expressed on tumor cells.

在一些實施例中，用於本文所提供之方法中之ADC包含與細胞毒性劑結合之抗體，其中抗體特異性結合選自以下之抗原：5T4 (TPBG)、ADAM-9 、AG-7、ALK、ALP、AMHRII、APLP2、ASCT2、AVB6、AXL (UFO)、B7-H3 (CD276)、B7-H4、BCMA、C3a、C3b、C4.4a (LYPD3)、C5、C5a、CA6、CA9、CanAg、碳酸酐酶IX (CAIX)、組織蛋白酶D、CCR7、CD1、CD10、CD100、CD101、CD102、CD103、CD104、CD105、CD106、CD107a、CD107b、CD108、CD109、CD111、CD112、CD113、CD116、CD117、CD118、CD119、CD11A、CD11b、CD11c、CD120a、CD121a、CD121b、CD122、CD123、CD124、CD125、CD126、CD127、CD13、CD130、CD131、CD132、CD133、CD135、CD136、CD137、CD138、CD14、CD140a、CD140b、CD141、CD142、CD143、CD144、CD146、CD147、CD148、CD15、CD150、CD151、CD154、CD155、CD156a、CD156b、CD156c、CD157、CD158b2、CD158e、CD158f1、CD158h、CD158i、CD159a、CD16、CD160、CD161、CD162、CD163、CD164、CD166、CD167b、CD169、CD16a、CD16b、CD170、CD171、CD172a、CD172b、CD172g、CD18、CD180、CD181、CD183、CD184、CD185、CD19、CD194、CD197、CD1a、CD1b、CD1c、CD1d、CD2、CD20、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD208、CD21、CD213a1、CD213a2、CD217、CD218a、CD22、CD220、CD221、CD222、CD224、CD226、CD228、CD229、CD23、CD230、CD232、CD239、CD243、CD244、CD248、CD249、CD25、CD26、CD265、CD267、CD269、CD27、CD272、CD273、CD274、CD275、CD279、CD28、CD280、CD281、CD282、CD283、CD284、CD289、CD29、CD294、CD295、CD298、CD3、CD3 ε、CD30、CD300f、CD302、CD304、CD305、CD307、CD31、CD312、CD315、CD316、CD317、CD318、CD319、CD32、CD321、CD322、CD324、CD325、CD326、CD327、CD328、CD32b、CD33、CD331、CD332、CD333、CD334、CD337、CD339、CD34、CD340、CD344、CD35、CD352、CD36、CD37、CD38、CD39、CD3d、CD3g、CD4、CD41、CD42d、CD44、CD44v6、CD45、CD46、CD47、CD48、CD49a、CD49b、CD49c、CD49d、CD49e、CD49f、CD5、CD50、CD51、CD51 (整合素α-V)、CD52、CD53、CD54、CD55、CD56、CD58、CD59、CD6、CD61、CD62L、CD62P、CD63、CD64、CD66a-e、CD67、CD68、CD69、CD7、CD70、CD70L、CD71、CD71 (TfR)、CD72、CD73、CD74、CD79a、CD79b、CD8、CD80、CD82、CD83、CD84、CD85f、CD85i、CD85j、CD86、CD87、CD89、CD90、CD91、CD92、CD95、CD96、CD97、CD98、CDH6、CDH6 (鈣黏素6)、CDw210a、CDw210b、CEA、CEACAM5、CEACAM6、CFC1B、cKIT、CLDN18.2 (密連蛋白18.2)、CLDN6、CLDN9、CLL-1、c-MET、補體因子C3、Cripto、CSP-1、CXCR5、DCLK1、DLK-1、DLL3、DPEP3、DR5 (死亡受體5)、抗黏附素、EFNA4、EGFR、EGFR野生型、EGFRviii、EGP-1 (TROP-2)、EGP-2、EMP2、ENPP3、EpCAM、EphA2、EphA3、Ephrin-A4 (EFNA4)、ETBR、FAP、FcRH5、FGFR2、FGFR3、FLT3、FOLR、FOLR1、FOLR-α、FSH、GCC、GD2、GD3、globo H、GPC1、GPC-1、GPC3、GPNMB、GPR20、HER2、HER-2、HER3、HER-3、HGFR (c-Met)、HLA-DR、HM1.24、HSP90、Ia、IGF-1R、IL-13R、IL-15、IL1RAP、IL-2、IL-3、IL-4、IL7R、整合素αVβ3、整合素β-6、介白素-4受體(IL4R)、KAAG-1、KLK2、LAMP-1、Le(y)、路易斯Y抗原、LGALS3BP、LGR5、LH/hCG、LHRH、脂筏、LIV-1 (SLC39A6或ZIP6)、LRP-1、LRRC15、LY6E、巨噬細胞甘露糖受體1、MAGE、間皮素(MSLN)、MET、I類MHC鏈相關蛋白A及B (MICA及MICB)、MN/CA IX、MRC2、MT1-MMP、MTX3、MTX5、MUC1、MUC16、MUC2、MUC3、MUC4、MUC5、MUC5ac、NaPi2b、NCA-90、NCA-95、連接素-4、Notch3、核仁素、OAcGD2、OT-MUC1 (腫瘤栓繫MUC1)、OX001L、P1GF、PAM4抗原、p-鈣黏素(鈣黏素3)、PD-L1、磷脂醯基絲胺酸(PS)、PRLR、促乳素受體(PRLR)、假單胞菌(Pseudomonas)、PSMA、PTK4、PTK7、受體酪胺酸激酶(RTK)、RNF43、ROR1、ROR2、SAIL、SEZ6、SLAMF7、SLC44A4、SLITRK6、SLMAMF7 (CS1)、SLTRK6、分選蛋白(SORT1)、SSEA-4、SSTR2、金黃色葡萄球菌(抗生素劑)、STEAP-1、STING、STn、T101、TAA、TAC、TDGF1、肌腱蛋白、TENB2、TGF-B、Thomson-Friedenreich抗原、Thy1.1、TIM-1、組織因子(TF；CD142)、TM4SF1、Tn抗原、TNF-alpha (TNFα)、TRA-1-60、TRAIL受體(R1及R2)、TROP-2、腫瘤相關糖蛋白72 (TAG-72)、uPAR、VEGFR、VEGFR-2及xCT。In some embodiments, the ADC used in the methods provided herein comprises an antibody that binds to a cytotoxic agent, wherein the antibody specifically binds an antigen selected from the group consisting of: 5T4 (TPBG), ADAM-9, AG-7, ALK , ALP, AMHRII, APLP2, ASCT2, AVB6, AXL (UFO), B7-H3 (CD276), B7-H4, BCMA, C3a, C3b, C4.4a (LYPD3), C5, C5a, CA6, CA9, CanAg, Carbonic Anhydrase IX (CAIX), Cathepsin D, CCR7, CD1, CD10, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107a, CD107b, CD108, CD109, CD111, CD112, CD113, CD116, CD117, CD118, CD119, CD11A, CD11b, CD11c, CD120a, CD121a, CD121b, CD122, CD123, CD124, CD125, CD126, CD127, CD13, CD130, CD131, CD132, CD133, CD135, CD136, CD137, CD138, CD14, CD140a CD140b, CD141, CD142, CD143, CD144, CD146, CD147, CD148, CD15, CD150, CD151, CD154, CD155, CD156a, CD156b, CD156c, CD157, CD158b2, CD158e, CD158f1, CD158h, CD158i, CD159a, CD158i CD161, CD162, CD163, CD164, CD166, CD167b, CD169, CD16a, CD16b, CD170, CD171, CD172a, CD172b, CD172g, CD18, CD180, CD181, CD183, CD184, CD185, CD19, CD194, CD197, CD1a, CD1b, CD1c, CD1d, CD2, CD20, CD200, CD201, CD202b, CD203c, CD204, CD205, CD206, CD208, CD21, CD213a1, CD213a2, CD217, CD218a, CD22, CD220, CD221, CD222, CD224, CD226, CD228, CD229, CD23, CD230, CD232, CD239, CD243, CD244, CD248, CD249, CD25, CD26, CD265, CD26 7. CD269, CD27, CD272, CD273, CD274, CD275, CD279, CD28, CD280, CD281, CD282, CD283, CD284, CD289, CD29, CD294, CD295, CD298, CD3, CD3ε, CD30, CD300f, CD302, CD304 , CD305, CD307, CD31, CD312, CD315, CD316, CD317, CD318, CD319, CD32, CD321, CD322, CD324, CD325, CD326, CD327, CD328, CD32b, CD33, CD331, CD332, CD333, CD334, CD337, CD339 , CD34, CD340, CD344, CD35, CD352, CD36, CD37, CD38, CD39, CD3d, CD3g, CD4, CD41, CD42d, CD44, CD44v6, CD45, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e , CD49f, CD5, CD50, CD51, CD51 (integrin alpha-V), CD52, CD53, CD54, CD55, CD56, CD58, CD59, CD6, CD61, CD62L, CD62P, CD63, CD64, CD66a-e, CD67, CD68, CD69, CD7, CD70, CD70L, CD71, CD71 (TfR), CD72, CD73, CD74, CD79a, CD79b, CD8, CD80, CD82, CD83, CD84, CD85f, CD85i, CD85j, CD86, CD87, CD89, CD90 , CD91, CD92, CD95, CD96, CD97, CD98, CDH6, CDH6 (Cadherin 6), CDw210a, CDw210b, CEA, CEACAM5, CEACAM6, CFC1B, cKIT, CLDN18.2 (claudin 18.2), CLDN6, CLDN9 , CLL-1, c-MET, complement factor C3, Cripto, CSP-1, CXCR5, DCLK1, DLK-1, DLL3, DPEP3, DR5 (death receptor 5), antiadhesin, EFNA4, EGFR, EGFR wild type , EGFRviii, EGP-1 (TROP-2), EGP-2, EMP2, ENPP3, EpCAM, EphA2, EphA3, Ephrin-A4 (EFNA4), ETBR, FAP, FcRH5, FGFR2, FGFR3, FLT3, FO LR, FOLR1, FOLR-α, FSH, GCC, GD2, GD3, globo H, GPC1, GPC-1, GPC3, GPNMB, GPR20, HER2, HER-2, HER3, HER-3, HGFR (c-Met), HLA-DR, HM1.24, HSP90, Ia, IGF-1R, IL-13R, IL-15, IL1RAP, IL-2, IL-3, IL-4, IL7R, integrin αVβ3, integrin β-6, Interleukin-4 receptor (IL4R), KAAG-1, KLK2, LAMP-1, Le(y), Lewis Y antigen, LGALS3BP, LGR5, LH/hCG, LHRH, lipid rafts, LIV-1 (SLC39A6 or ZIP6 ), LRP-1, LRRC15, LY6E, macrophage mannose receptor 1, MAGE, mesothelin (MSLN), MET, class I MHC chain-associated proteins A and B (MICA and MICB), MN/CA IX, MRC2, MT1-MMP, MTX3, MTX5, MUC1, MUC16, MUC2, MUC3, MUC4, MUC5, MUC5ac, NaPi2b, NCA-90, NCA-95, Connexin-4, Notch3, Nucleolin, OAcGD2, OT-MUC1 (Tumor tethered MUC1), OX001L, P1GF, PAM4 antigen, p-cadherin (Cadherin 3), PD-L1, phosphatidylserine (PS), PRLR, prolactin receptor (PRLR) , Pseudomonas, PSMA, PTK4, PTK7, receptor tyrosine kinase (RTK), RNF43, ROR1, ROR2, SAIL, SEZ6, SLAMF7, SLC44A4, SLITRK6, SLMAMF7 (CS1), SLTRK6, sorting protein (SORT1), SSEA-4, SSTR2, Staphylococcus aureus (antibiotic agent), STEAP-1, STING, STn, T101, TAA, TAC, TDGF1, tenascin, TENB2, TGF-B, Thomson-Friedenreich antigen, Thy1.1, TIM-1, tissue factor (TF; CD142), TM4SF1, Tn antigen, TNF-alpha (TNFα), TRA-1-60, TRAIL receptors (R1 and R2), TROP-2, tumor-associated carbohydrates Protein 72 (TAG-72), uPAR, VEGFR, VEGFR-2 and xCT.

在一些實施例中，ADC結合選自以下之抗原：EGFR、KAAG1、MET、CD30、HER2、CD30、IL7R、CD248、腫瘤相關糖蛋白72 (TAG-72)、MRC2、EGFR、CD71、TRA-1-60、STn、CLDN18.2、CLDN6、HER-2、CD33、CD7、OT-MUC1 (腫瘤栓繫MUC1)、TRA-1-60、TIM-1、GCC、間皮素(MSLN)、EGFR、gpNMB、CD20、AMHRII、NaPi2b、CD142、ROR1、整合素β6、Ly6E、cMET、CD37、MUC16、STEAP-1、LRRC15、SLITRK6、MUC16、ETBR、FCRH5、Axl、CD79b、Globo H、SLAMF7、PSMA、CD22、CD228、CD48、LIV-1、EphA2、SLC44A4、CA9、Axl及LGR5。In some embodiments, the ADC binds to an antigen selected from the group consisting of: EGFR, KAAG1, MET, CD30, HER2, CD30, IL7R, CD248, tumor-associated glycoprotein 72 (TAG-72), MRC2, EGFR, CD71, TRA-1 -60, STn, CLDN18.2, CLDN6, HER-2, CD33, CD7, OT-MUC1 (tumor tethered MUC1), TRA-1-60, TIM-1, GCC, mesothelin (MSLN), EGFR, gpNMB, CD20, AMHRII, NaPi2b, CD142, ROR1, integrin beta6, Ly6E, cMET, CD37, MUC16, STEAP-1, LRRC15, SLITRK6, MUC16, ETBR, FCRH5, Axl, CD79b, Globo H, SLAMF7, PSMA, CD22 , CD228, CD48, LIV-1, EphA2, SLC44A4, CA9, Axl and LGR5.

在一些實施例中，ADC結合選自以下之抗原：BCMA、GPC1、CD30、cMET、SAIL、HER3、CD70、c-MET、CD46、HER2、5T4、ENPP3、CD19、EGFR、BCMA、CD70、BCMA及EphA2。In some embodiments, the ADC binds to an antigen selected from the group consisting of BCMA, GPC1, CD30, cMET, SAIL, HER3, CD70, c-MET, CD46, HER2, 5T4, ENPP3, CD19, EGFR, BCMA, CD70, BCMA, and EphA2.

在一些實施例中，ADC結合選自以下之抗原：Her2、TROP2、BCMA、cMet、整合素αVβ6、CD22、CD79b、CD30、CD19、CD70、CD228及CD47。In some embodiments, the ADC binds an antigen selected from the group consisting of Her2, TROP2, BCMA, cMet, integrin αVβ6, CD22, CD79b, CD30, CD19, CD70, CD228, and CD47.

在一些實施例中，ADC結合選自以下之抗原：CD142、整合素β-6 、ENPP3、CD19、Ly6E、cMET、C4.4a、CD37、MUC16、STEAP-1、LRRC15、SLITRK6、MUC16、ETBR、FCRH5、Axl、EGFR、CD79b、BCMA、CD70、PSMA、CD79b、CD228、CD48、LIV-1、EphA2、SLC44A4、CD30及sTn。In some embodiments, the ADC binds to an antigen selected from the group consisting of CD142, integrin beta-6, ENPP3, CD19, Ly6E, cMET, C4.4a, CD37, MUC16, STEAP-1, LRRC15, SLITRK6, MUC16, ETBR, FCRH5, Axl, EGFR, CD79b, BCMA, CD70, PSMA, CD79b, CD228, CD48, LIV-1, EphA2, SLC44A4, CD30 and sTn.

在一些實施例中，ADC結合選自以下之抗原：5T4、ADAM-9、AG-7、ALK、AMHRII、APLP2、ASCT2、Axl、B7-H3、B7-H4、BCMA、C4.4a、CA6、CA9、CanAg、碳酸酐酶IX (CAIX)、組織蛋白酶D、CCR7、CD103、CD123、CD133、CD138、CD142、CD147、CD16、CD166、CD184、CD19、CD20、CD205、CD206、CD22、CD228、CD248、CD25、CD3、CD3 ε、CD30、CD300f、CD317、CD33、CD352、CD37、CD38、CD44v6、CD45、CD46、CD47、CD48、CD51、CD56、CD7、CD70、CD71、CD74、CD79b、CDH6、CEA、CEACAM5、CEACAM6、cKIT、CLDN18.2、CLDN6、CLDN9、CLL-1、c-MET、Cripto、CSP-1、CXCR5、DCLK1、DLK-1、DLL3、DPEP3、DR5 (死亡受體5)、抗黏附素、EFNA4、EGFR、EGFR野生型、EGFRviii、EMP2、ENPP3、EpCAM、EphA2、EphA3、ETBR、FAP、FCRH5、FGFR2、FGFR3、FLT3、FOLR、FOLR-α、FSH、GCC、GD2、GD3、Globo H、GPC-1、GPC3、gpNMB、GPR20、HER-2、HER-3、HLA-DR、HSP90、IGF-1R、IL-13R、IL-15、IL1RAP、IL-2、IL-3、IL-4、IL7R、整合素β-6、介白素-4受體(IL4R)、KAAG-1、KLK2、LAMP-1、路易斯Y抗原、LGALS3BP、LGR5、LH/hCG、LHRH、脂筏、LIV-1、LRP-1、LRRC15、Ly6E、巨噬細胞甘露糖受體1、MAGE、間皮素(MSLN)、MET、I類MHC鏈相關蛋白A及B (MICA及MICB)、MRC2、MT1-MMP、MTX3、MTX5、MUC-1、MUC16、NaPi2b、連接素-4、NOTCH3、核仁素、OAcGD2、OT-MUC1 (腫瘤栓繫MUC1)、OX001L、P-鈣黏素、PD-L1、磷脂醯基絲胺酸、磷脂醯絲胺酸(PS)、促乳素受體(PRLR)、假單胞菌屬、PSMA、PTK7、受體酪胺酸激酶(RTK)、RNF43、ROR1、ROR2、SAIL、SEZ6、SLAMF7、SLC44A4、SLITRK6、分選蛋白(SORT1)、SSEA-4、SSTR2、金黃色葡萄球菌(抗生素藥劑)、STEAP-1、STING、STING (有效負載目標)、STn、TAA、TGF-B、TIM-1、TM4SF1、TNF-α、TRA-1-60、TROP-2、腫瘤相關糖蛋白72 (TAG-72)、VEGFR-2、xCT。In some embodiments, the ADC binds to an antigen selected from the group consisting of: 5T4, ADAM-9, AG-7, ALK, AMHRII, APLP2, ASCT2, Axl, B7-H3, B7-H4, BCMA, C4.4a, CA6, CA9, CanAg, Carbonic Anhydrase IX (CAIX), Cathepsin D, CCR7, CD103, CD123, CD133, CD138, CD142, CD147, CD16, CD166, CD184, CD19, CD20, CD205, CD206, CD22, CD228, CD248, CD25, CD3, CD3 epsilon, CD30, CD300f, CD317, CD33, CD352, CD37, CD38, CD44v6, CD45, CD46, CD47, CD48, CD51, CD56, CD7, CD70, CD71, CD74, CD79b, CDH6, CEA, CEACAM5 , CEACAM6, cKIT, CLDN18.2, CLDN6, CLDN9, CLL-1, c-MET, Cripto, CSP-1, CXCR5, DCLK1, DLK-1, DLL3, DPEP3, DR5 (death receptor 5), antiadhesin , EFNA4, EGFR, EGFR wild type, EGFRviii, EMP2, ENPP3, EpCAM, EphA2, EphA3, ETBR, FAP, FCRH5, FGFR2, FGFR3, FLT3, FOLR, FOLR-α, FSH, GCC, GD2, GD3, Globo H, GPC-1, GPC3, gpNMB, GPR20, HER-2, HER-3, HLA-DR, HSP90, IGF-1R, IL-13R, IL-15, IL1RAP, IL-2, IL-3, IL-4, IL7R, integrin beta-6, interleukin-4 receptor (IL4R), KAAG-1, KLK2, LAMP-1, Lewis Y antigen, LGALS3BP, LGR5, LH/hCG, LHRH, lipid rafts, LIV-1, LRP-1, LRRC15, Ly6E, macrophage mannose receptor 1, MAGE, mesothelin (MSLN), MET, class I MHC chain-associated proteins A and B (MICA and MICB), MRC2, MT1-MMP, MTX3 , MTX5, MUC-1, MUC16, NaPi2b, connexin-4, NOTCH3, nucleolin, OAcGD2, OT-MUC1 (tumor tethered MUC1), OX001L, P-cadherin, PD-L1, phospholipids Amino Acid, Phosphatidylserine (PS), Prolactin Receptor (PRLR), Pseudomonas, PSMA, PTK7, Receptor Tyrosine Kinase (RTK), RNF 43, ROR1, ROR2, SAIL, SEZ6, SLAMF7, SLC44A4, SLITRK6, Sortin (SORT1), SSEA-4, SSTR2, S. aureus (antibiotic agent), STEAP-1, STING, STING (payload target) , STn, TAA, TGF-B, TIM-1, TM4SF1, TNF-α, TRA-1-60, TROP-2, tumor-associated glycoprotein 72 (TAG-72), VEGFR-2, xCT.

在一些實施例中，ADC結合選自以下之抗原：AMHRII、Axl、CA9、CD142、CD20、CD22、CD228、CD248、CD30、CD33、CD7、CD48、CD71、CD79b、CLDN18.2、CLDN6、c-MET、EGFR、EphA2、ETBR、FCRH5、GCC、Globo H、gpNMB、HER-2、IL7R、整合素β-6、KAAG-1、LGR5、LIV-1、LRRC15、Ly6E、間皮素(MSLN)、MET、MRC2、MUC16、NaPi2b、連接素-4、OT-MUC1 (腫瘤栓繫MUC1)、PSMA、ROR1、SLAMF7、SLC44A4、SLITRK6、STEAP-1、STn、TIM-1、TRA-1-60、腫瘤相關糖蛋白72 (TAG-72)。In some embodiments, the ADC binds an antigen selected from AMHRII, Axl, CA9, CD142, CD20, CD22, CD228, CD248, CD30, CD33, CD7, CD48, CD71, CD79b, CLDN18.2, CLDN6, c- MET, EGFR, EphA2, ETBR, FCRH5, GCC, Globo H, gpNMB, HER-2, IL7R, integrin beta-6, KAAG-1, LGR5, LIV-1, LRRC15, Ly6E, mesothelin (MSLN), MET, MRC2, MUC16, NaPi2b, connexin-4, OT-MUC1 (tumor tethered MUC1), PSMA, ROR1, SLAMF7, SLC44A4, SLITRK6, STEAP-1, STn, TIM-1, TRA-1-60, tumor Associated glycoprotein 72 (TAG-72).

在一些實施例中，ADC結合選自以下之抗原：BCMA、GPC-1、CD30、c-MET、SAIL、HER-3、CD70、CD46、HER-2、5T4、ENPP3、CD19、EGFR、EphA2。In some embodiments, the ADC binds to an antigen selected from the group consisting of BCMA, GPC-1, CD30, c-MET, SAIL, HER-3, CD70, CD46, HER-2, 5T4, ENPP3, CD19, EGFR, EphA2.

在一些實施例中，ADC之抗體不結合連接素-4。In some embodiments, the antibody to the ADC does not bind connexin-4.

通常，ADC之抗體及結合免疫細胞接合子之抗體為兩種獨立抗體。然而，在某些實施例中，抗體可形成雙特異性抗體。 B. 例示性細胞毒素劑 Typically, the antibody to the ADC and the antibody that binds to the immune cell engager are two separate antibodies. However, in certain embodiments, the antibodies may form bispecific antibodies. B. Exemplary Cytotoxic Agents

在各種實施例中，本文所提供之方法包含投與抗體-藥物結合物，其中抗體-藥物結合物包含與微管蛋白干擾劑結合之抗體。In various embodiments, the methods provided herein comprise administering an antibody-drug conjugate, wherein the antibody-drug conjugate comprises an antibody that binds to a tubulin interfering agent.

本領域已知各種類別之微管蛋白干擾劑，包括但不限於尾海兔素、奧瑞他汀、微管蛋白裂解素、秋水仙鹼、長春花生物鹼、紫杉烷、T67 (Tularik)、克瑞托欣、類美登素、哈米特林及其他微管蛋白干擾劑。Various classes of tubulin interfering agents are known in the art, including, but not limited to, Aplysin, Auristatin, Tubulin Lysin, Colchicine, Vinca alkaloids, taxanes, T67 (Tularik), Cratosin, maytansinoids, hamitrin, and other tubulin-disrupting agents.

奧瑞他汀為天然產物尾海兔素之衍生物。例示性奧瑞他汀包括尾海兔素-10、奧瑞斯他汀E、奧瑞他汀T、MMAE (N-甲基纈胺酸-纈胺酸-多拉索因-多拉普因-降麻黃鹼或單甲基奧瑞他汀E)及MMAF (N-甲基纈胺酸-纈胺酸-多拉索因-多拉普因-苯丙胺酸或多纈胺酸(dovaline)-纈胺酸-多拉索因-多拉普因-苯丙胺酸)、AEB (藉由使奧瑞斯他汀E與對乙醯基苯甲酸反應產生的酯)、AEVB (藉由使奧瑞斯他汀E與苯甲醯基戊酸反應產生的酯)及AFP (二甲基纈胺酸-纈胺酸-多拉索因-多拉普因-苯丙胺酸-對苯二胺或奧瑞他汀苯丙胺酸苯二胺)。WO 2015/057699描述了PEG化之奧瑞他汀，包括MMAE。考慮使用之另外的尾海兔素衍生物揭示於美國專利第9,345,785號中，其出於任何目的以引入之方式併入本文中。例示性奧瑞他汀實施例包括N末端鍵聯之單甲基奧瑞他汀藥物單元DE及DF，揭示於2004年3月28日提交之「Senter等人，Proceedings of the American Association for Cancer Research, 第45卷, 摘要編號623中，且在美國專利公開案第2005/0238649號中有描述，該專利之揭示內容明確地以全文引用之方式併入。Auristatin is a derivative of the natural product Dolastatin. Exemplary auristatins include dolastatin-10, auristatin E, auristatin T, MMAE (N-methylvaline-valine-dorasoine-drapine-norestatin) Flaline or monomethylauristatin E) and MMAF (N-methylvaline-valine-dorasoine-dorapine-phenylalanine or dovaline-valine) - doraxoin-dorapine-phenylalanine), AEB (ester produced by reacting auristatin E with p-acetylbenzoic acid), AEVB (by reacting auristatin E with benzene Esters produced by the reaction of methylvaleric acid) and AFP (dimethylvaline-valine-dorasoine-dorapine-phenylalanine-p-phenylenediamine or auristatin-phenylalanine-phenylenediamine) ). WO 2015/057699 describes PEGylated auristatins, including MMAE. Additional dolastatin derivatives contemplated for use are disclosed in US Patent No. 9,345,785, which is incorporated herein by reference for any purpose. Exemplary auristatin examples include N-terminally linked monomethyl auristatin drug units DE and DF disclosed in "Senter et al., Proceedings of the American Association for Cancer Research, vol. 45, Abstract No. 623, and described in US Patent Publication No. 2005/0238649, the disclosure of which is expressly incorporated by reference in its entirety.

在某些實施例中，ADC細胞毒性劑為MMAE。In certain embodiments, the ADC cytotoxic agent is MMAE.

在其他實施例中，與ADC結合之細胞毒性劑為MMAF。In other embodiments, the cytotoxic agent conjugated to the ADC is MMAF.

微管蛋白裂解素包括但不限於微管蛋白裂解素D、微管蛋白裂解素M、微管丙胺酸及微管酪胺酸。WO 2017-096311及WO 2016-040684描述了非限制性微管蛋白裂解素類似物，包括微管蛋白裂解素M。Tubulins include, but are not limited to, tubulin D, tubulin M, tubulin alanine, and tubulin tyrosine. WO 2017-096311 and WO 2016-040684 describe non-limiting tubulin analogs, including tubulin M.

秋水仙鹼包括但不限於秋水仙鹼及CA-4。Colchicine includes, but is not limited to, colchicine and CA-4.

長春花生物鹼包括但不限於長春鹼(VBL)、長春瑞賓(vinorelbine；VRL)、長春新鹼(vincristine；VCR)及長春地辛(vindfesine；VDS)。Vinca alkaloids include, but are not limited to, vinblastine (VBL), vinorelbine (VRL), vincristine (VCR), and vindfesine (VDS).

紫杉烷包括但不限於Taxol ®(太平洋紫杉醇)及Taxotere ®(多西他賽(docetaxel))。 Taxanes include, but are not limited to, Taxol® (paclitaxel) and Taxotere® (docetaxel).

克瑞托欣包括但不限於克瑞托欣-1及克瑞托欣-52。Kratoshin includes, but is not limited to, Kratoshin-1 and Kratoshin-52.

類美登素包括但不限於美登素、美登醇、美登素類似物、DM1、DM3及DM4以及安莎黴素(ansamatocin)-2。例示性類美登素藥物部分包括具有經修飾之芳環的彼等類美登素藥物部分，諸如：C-19-去氯(美國專利第4,256,746號) (藉由氫化鋰鋁還原安絲菌素(ansamytocin) P2來製備)；C-20-羥基(或C-20-去甲基) +/-C-19-去氯(美國專利第4,361,650號及第4,307,016號) (藉由使用鏈黴菌屬(Streptomyces)或放線菌屬(Actinomyces)去甲基化或使用LAH去氯來製備)；以及C-20-去甲氧基、C-20-醯氧基(OCOR)、+/-去氯(美國專利第4,294,757號) (藉由使用醯基氯進行醯化來製備)及在其他位置具有修飾之彼等類美登素藥物部分。Maytansinoids include, but are not limited to, maytansine, maytansinol, maytansine analogs, DM1, DM3, and DM4, and ansamatocin-2. Exemplary maytansinoid drug moieties include those with modified aromatic rings, such as: C-19-Dechloro (US Pat. No. 4,256,746) (Reduction of A. monocytogenes by lithium aluminum hydride) ansamytocin P2); C-20-hydroxy (or C-20-demethyl) +/- C-19-dechloro (US Pat. Nos. 4,361,650 and 4,307,016) (by using Streptomyces sp. Streptomyces or Actinomyces demethylated or prepared using LAH dechlorination); and C-20-demethoxy, C-20-oxo (OCOR), +/- dechlorination (US Pat. No. 4,294,757) (prepared by acylation using acyl chloride) and their maytansinoid drug moieties with modifications at other positions.

類美登素藥物部分亦包括具有以下修飾之彼等類美登素藥物部分，諸如：C-9-SH(美國專利第4,424,219號) (藉由使美登醇與H.sub.25或P.sub.2S.sub.5反應來製備)；C-14-烷氧基甲基(去甲氧基/CH.sub.20R) (美國專利第4,331,598號)；C-14-羥基甲基或醯氧基甲基(CH.sub.20H或CH.sub.2OAc) (美國專利第4,450,254號) (由諾卡菌屬(Nocardia)製備)；C-15-羥基/醯氧基(美國專利第4,364,866號) (藉由用鏈黴菌轉化美登醇來製備)；C-15-甲氧基(美國專利第4,313,946號及第4,315,929號) (自滑桃樹(Trewia nudlflora)分離)；C-18-N-去甲基(美國專利第4,362,663號及第4,322,348號) (藉由用鏈黴菌使美登醇去甲基化來製備)；及4,5-去氧(美國專利第4,371,533號) (藉由三氯化鈦/LAH還原美登醇來製備)。在人類乳癌細胞株SK-BR-3上活體外測試結合HER-2之TA.1-類美登素結合物的細胞毒性(Chari等人，Cancer Research 52:127-131 (1992))。藥物結合物達成類似於游離類美登素藥物之細胞毒性程度，其可藉由增加每個抗體分子之類美登素分子數目而增加。Maytansinoid drug moieties also include those maytansinoid drug moieties with modifications such as: C-9-SH (US Pat. No. 4,424,219) (by combining maytansinol with H.sub.25 or P .sub.2S.sub.5 reaction); C-14-alkoxymethyl (demethoxy/CH.sub.20R) (US Patent No. 4,331,598); C-14-hydroxymethyl or Ethyloxymethyl (CH.sub.20H or CH.sub.2OAc) (US Pat. No. 4,450,254) (prepared by Nocardia); C-15-Hydroxy/Acidyloxy (US Pat. 4,364,866) (prepared by transforming maytansinol with Streptomyces sp.); C-15-methoxy (US Pat. Nos. 4,313,946 and 4,315,929) (isolated from Trewia nudlflora); C-18 -N-Demethyl (US Pat. Nos. 4,362,663 and 4,322,348) (prepared by demethylation of maytansinol with Streptomyces sp.); and 4,5-Deoxy (US Pat. No. 4,371,533) ( Prepared by reduction of maytansinol with titanium trichloride/LAH). The cytotoxicity of HER-2-binding TA.1-maytansinoid conjugates was tested in vitro on the human breast cancer cell line SK-BR-3 (Chari et al., Cancer Research 52:127-131 (1992)). The drug conjugate achieves a degree of cytotoxicity similar to that of the free maytansinoid drug, which can be increased by increasing the number of maytansinoid molecules per antibody molecule.

哈米特林包括但不限於哈米特林及HTI-286。Hamitlin includes, but is not limited to, Hamitlin and HTI-286.

其他微管蛋白干擾劑包括塔卡醯胺(taccalonolide) A、塔卡醯胺B、塔卡醯胺AF、塔卡醯胺AJ、塔卡醯胺AI-環氧化物、圓皮海綿內酯(discodermolide)、巴卡丁衍生物(baccatin derivatives)、紫杉烷類似物(例如埃坡黴素(epothilone) A及埃坡黴素B)、諾考達唑(nocodazole)、秋水仙鹼、秋水醯胺、雌氮芥(estramustine)、西馬多丁(cemadotin)、風車子抑素(combretastatins)、圓皮海綿內酯、艾榴塞洛素(eleutherobin)、艾日布林(eribulin)、拉博林(prolabolin)、福莫普辛(phomopsin)及萊利黴素(laulimalide)。Other tubulin-disrupting agents include taccalonolide A, taccalonolide B, taccalonolide AF, taccalonolide AJ, taccalonolide AI-epoxide, spongolide ( discodermolide), baccatin derivatives, taxane analogs (eg epothilone A and epothilone B), nocodazole, colchicine, colchicine Amine, estramustine, cemadotin, combretastatins, spongolide, eleutherobin, eribulin, labo Lin (prolabolin), formopsin (phomopsin) and leilimycin (laulimalide).

在一些實施例中，用於本文中之方法的ADC可包含連接子單元。舉例而言，ADC可包含細胞毒性劑與抗體之間的連接子區。在一些實施例中，連接子為蛋白酶可裂解連接子、酸可裂解連接子、二硫化物連接子或自穩定連接子。在各種實施例中，連接子可在細胞內條件下裂解，使得連接子之裂解自細胞內環境中之抗體釋放治療劑。In some embodiments, ADCs used in the methods herein may include linker units. For example, the ADC can comprise a linker region between the cytotoxic agent and the antibody. In some embodiments, the linker is a protease-cleavable linker, an acid-cleavable linker, a disulfide linker, or a self-stabilizing linker. In various embodiments, the linker is cleavable under intracellular conditions such that cleavage of the linker releases the therapeutic agent from the antibody in the intracellular environment.

用於本文中之方法的ADC可包含連接子，其中治療劑(例如微管蛋白干擾劑)可以降低抗體活性的方式與抗體結合，除非該連接子自抗體剝落(例如藉由水解、藉由抗體降解或藉由裂解劑)。此類治療劑可經由連接子而連接至抗體。與連接子結合之治療劑在本文中亦稱作藥物連接子。連接子之性質可廣泛變化。構成連接子之組分係基於其特徵加以選擇，其可部分地根據結合物所遞送之位點處的條件來指定。ADCs used in the methods herein can include a linker, wherein a therapeutic agent (eg, a tubulin interfering agent) can bind to the antibody in a manner that reduces the activity of the antibody, unless the linker is exfoliated from the antibody (eg, by hydrolysis, by the antibody degradation or by lysing agents). Such therapeutic agents can be linked to the antibody via a linker. A therapeutic agent bound to a linker is also referred to herein as a drug linker. The nature of the linker can vary widely. The components that make up the linker are selected based on their characteristics, which can be specified in part by the conditions at the site where the conjugate is delivered.

治療劑可藉由對目標細胞之細胞內環境中裂解敏感但對細胞外環境大體上不敏感之可裂解連接子與抗體連接，使得結合物在其藉由癌細胞內化(例如在核內體中，或例如在溶酶體環境或胞膜窖環境(caveolear environment)中藉助於pH敏感性或蛋白酶敏感性)時自抗體裂解。治療劑亦可藉由不可裂解連接子與抗體連接。The therapeutic agent can be linked to the antibody via a cleavable linker that is sensitive to cleavage in the intracellular environment of the target cell but is substantially insensitive to the extracellular environment, allowing the conjugate to be internalized by the cancer cell (e.g., in endosomes). cleavage from the antibody, or, for example, in a lysosomal environment or caveolear environment by means of pH sensitivity or protease sensitivity. The therapeutic agent can also be linked to the antibody via a non-cleavable linker.

如所指示，連接子可包含可裂解單元。在一些此類實施例中，可裂解單元之結構及/或序列經選定，使得其藉由在目標位點(例如目標細胞)處存在之酶作用而裂解。在其他實施例中，亦可使用可藉由pH(例如酸或鹼不穩定)、溫度變化或在輻照(例如光不穩定)時裂解之可裂解單元。As indicated, the linker may comprise a cleavable unit. In some such embodiments, the structure and/or sequence of the cleavable unit is selected such that it is cleaved by the action of an enzyme present at the target site (eg, the target cell). In other embodiments, cleavable units that can be cleaved by pH (eg, acid or base labile), temperature changes, or upon irradiation (eg, photolabile) can also be used.

在一些實施例中，可裂解單元可包含一個胺基酸或胺基酸之連續序列。胺基酸序列可為酶之目標受質。In some embodiments, the cleavable unit may comprise an amino acid or a contiguous sequence of amino acids. The amino acid sequence can be the target substrate for the enzyme.

在一些態樣中，可裂解單元為肽基單元且為至少兩個胺基酸長。裂解劑可包括組織蛋白酶B及D及纖維蛋白溶酶(參見例如Dubowchik及Walker, 1999, Pharm. Therapeutics83:67-123)。最典型為可藉由目標細胞中存在之酶裂解的可裂解單元，亦即酶可裂解連接子。因此，連接子可例如藉由包括溶酶體或胞內體蛋白酶之細胞內肽酶或蛋白酶裂解。舉例而言，可使用可藉由在癌組織中高度表現之硫醇依賴性蛋白酶組織蛋白酶B裂解之連接子(例如包含Phe-Leu或Val-Cit肽或Val-Ala肽之連接子)。 In some aspects, the cleavable unit is a peptidyl unit and is at least two amino acids long. Cleavage agents can include cathepsins B and D and plasmin (see, eg, Dubowchik and Walker, 1999, Pharm. Therapeutics 83:67-123). Most typically a cleavable unit that is cleavable by enzymes present in the target cell, ie an enzyme cleavable linker. Thus, the linker can be cleaved, for example, by intracellular peptidases or proteases including lysosomal or endosome proteases. For example, linkers that can be cleaved by the thiol-dependent protease cathepsin B, which are highly expressed in cancer tissues (eg, linkers comprising Phe-Leu or Val-Cit peptides or Val-Ala peptides) can be used.

在一些實施例中，連接子將包含可裂解單元(例如肽基單元)且可裂解單元將直接與治療劑結合。在其他實施例中，可裂解單元將經由另外功能單元，例如自我分解型間隔子單元或非自我分解型間隔子單元與治療劑結合。非自我分解型間隔子單元為其中部分或全部間隔子單元在可裂解單元(例如胺基酸)自抗體-藥物結合物裂解之後保持與藥物單元結合的間隔子單元。為釋放藥物，在目標細胞內發生單獨水解反應以使間隔子單元自藥物裂解。In some embodiments, the linker will comprise a cleavable unit (eg, a peptidyl unit) and the cleavable unit will bind directly to the therapeutic agent. In other embodiments, the cleavable unit will bind to the therapeutic agent via an additional functional unit, such as a self-decomposing spacer unit or a non-self-decomposing spacer unit. A non-self-decomposing spacer unit is one in which some or all of the spacer unit remains bound to the drug unit after cleavage of the cleavable unit (eg, amino acid) from the antibody-drug conjugate. To release the drug, a separate hydrolysis reaction takes place within the target cell to cleave the spacer unit from the drug.

藉由自我分解型間隔子單元，藥物無需進行單獨水解步驟，而釋放藥物。在一個實施例中，其中連接子包含可裂解單元及自我分解型基團，可裂解單元可藉由酶之作用裂解且在可裂解單元裂解之後，自我分解型基團釋放治療劑。在一些實施例中，連接子之可裂解單元將在一端上直接地或間接地與治療劑結合而在另一端上將直接地或間接地與抗體結合。在一些此類實施例中，可裂解單元將在一端上直接地或間接地(例如經由自我分解型或非自我分解型間隔子單元)與治療劑結合而在另一端上將經由延伸子單元與抗體結合。延伸子單元將抗體連接至藥物及/或藥物連接子之剩餘部分。在一個實施例中，抗體與藥物或藥物連接子之剩餘部分之間的連接係經由順丁烯二醯亞胺基，例如經由順丁烯二醯亞胺基己醯基連接子。在一些實施例中，抗體將經由二硫化物鍵聯至藥物，例如二硫化物鍵聯之類美登素結合SPDB-DM4及SPP-DM1。With the self-decomposing spacer unit, the drug does not need to undergo a separate hydrolysis step to release the drug. In one embodiment, wherein the linker comprises a cleavable unit and a self-decomposing group, the cleavable unit is cleavable by the action of an enzyme and upon cleavage of the cleavable unit, the self-degrading group releases the therapeutic agent. In some embodiments, the cleavable unit of the linker will bind directly or indirectly to the therapeutic agent on one end and will bind directly or indirectly to the antibody on the other end. In some such embodiments, the cleavable unit will bind directly or indirectly (eg, via a self-decomposing or non-self-decomposing spacer unit) to the therapeutic agent on one end and will bind via an extension subunit on the other end Antibody binding. The extension subunit links the antibody to the drug and/or the remainder of the drug linker. In one embodiment, the linkage between the antibody and the drug or the remainder of the drug linker is via a maleimino group, eg, via a maleiminohexyl linker. In some embodiments, the antibody will be linked to the drug via a disulfide linkage, eg, a disulfide linkage like maytansine binds SPDB-DM4 and SPP-DM1.

抗體與連接子之間的連接可經由多種不同途徑，例如經由硫醚鍵、經由二硫鍵、經由醯胺鍵或經由酯鍵。在一個實施例中，抗體與連接子之間的連接形成於抗體之半胱胺酸殘基的硫醇基與連接子之順丁烯二醯亞胺基之間。在一些實施例中，抗體之鏈間鍵在與連接子之官能基反應之前轉化為游離硫醇基。在一些實施例中，半胱胺酸殘基經引入至抗體之重鏈或輕鏈中且與連接子反應。藉由抗體重鏈或輕鏈中之取代進行半胱胺酸插入之位置包括公開的美國申請案第2007-0092940號及國際專利公開案WO2008070593號中所描述之位置，該等參考中之每一者均以全文引用之方式且出於所有目的併入本文中。The linkage between the antibody and the linker can be via a number of different routes, such as via a thioether bond, via a disulfide bond, via an amide bond, or via an ester bond. In one embodiment, the linkage between the antibody and the linker is formed between the thiol group of the cysteine residue of the antibody and the maleimine group of the linker. In some embodiments, the interchain linkages of the antibodies are converted to free thiol groups prior to reaction with the functional groups of the linker. In some embodiments, a cysteine residue is introduced into the heavy or light chain of the antibody and reacted with a linker. Positions for cysteine insertion by substitution in the antibody heavy or light chain include those described in Published US Application No. 2007-0092940 and International Patent Publication No. WO2008070593, each of which references All are incorporated herein by reference in their entirety and for all purposes.

在一些實施例中，抗體-藥物結合物具有下式I： L - (LU-D) p(I) 其中L為抗體，LU為連接子單元且D為藥物單元(亦即治療劑)。下標p介於1至20範圍內。此類結合物包含經由連接子共價鍵聯至至少一種藥物之抗體。連接子單元在一端處連接至抗體而在另一端處連接至藥物。 In some embodiments, the antibody-drug conjugate has the following formula I: L-(LU-D) p (I) wherein L is the antibody, LU is the linker unit and D is the drug unit (ie, the therapeutic agent). The subscript p is in the range 1 to 20. Such conjugates comprise antibodies covalently linked to at least one drug via a linker. The linker unit is linked to the antibody at one end and to the drug at the other end.

藥物負載由p表示，其為每一抗體之藥物分子數目。藥物負載可介於1至20個藥物單元(D)/抗體範圍內。在一些態樣中，下標p將介於1至20範圍內(亦即1至20之整數值及非整數值兩者)。在一些態樣中，下標p將為1至20之整數，且將表示單一抗體上藥物-連接子的數目。在其他態樣中，p表示平均藥物-連接子分子數目/抗體，例如反應混合物或組合物(例如醫藥組合物)中平均藥物-連接子數目/抗體，且可為整數或非整數值。因此，在一些態樣中，對於組合物(例如醫藥組合物)，p表示組合物中抗體-藥物結合物之平均藥物負載，且p介於1至20範圍內。Drug loading is represented by p, which is the number of drug molecules per antibody. Drug loading can range from 1 to 20 drug units (D) per antibody. In some aspects, the subscript p will be in the range 1-20 (ie, both integer and non-integer values of 1-20). In some aspects, the subscript p will be an integer from 1 to 20, and will represent the number of drug-linkers on a single antibody. In other aspects, p represents the average number of drug-linker molecules per antibody, eg, the average number of drug-linkers per antibody in a reaction mixture or composition (eg, a pharmaceutical composition), and can be an integer or non-integer value. Thus, in some aspects, for a composition (eg, a pharmaceutical composition), p represents the average drug loading of the antibody-drug conjugate in the composition, and p ranges from 1 to 20.

在一些實施例中，p為約1至約8種藥物/抗體。在一些實施例中，p為1。在一些實施例中，p為2。在一些實施例中，p為約2至約8種藥物/抗體。在一些實施例中，p為約2至約6、2至約5或2至約4種藥物/抗體。在一些實施例中，p為約2、約4、約6或約8種藥物/抗體。In some embodiments, p is about 1 to about 8 drugs/antibody. In some embodiments, p is 1. In some embodiments, p is 2. In some embodiments, p is about 2 to about 8 drugs/antibody. In some embodiments, p is about 2 to about 6, 2 to about 5, or 2 to about 4 drugs/antibody. In some embodiments, p is about 2, about 4, about 6, or about 8 drugs/antibody.

在利用結合反應之製備中，平均藥物數目/抗體單元可藉由習知手段，諸如質譜法、ELISA分析、HIC及HPLC表徵。亦可就p而言來測定結合物之定量分佈。In preparations utilizing binding reactions, the average number of drug per antibody unit can be characterized by conventional means such as mass spectrometry, ELISA analysis, HIC and HPLC. The quantitative distribution of the conjugate can also be determined in terms of p.

例示性抗體-藥物結合物包括基於奧瑞他汀之抗體-藥物結合物，亦即其中藥物組分為奧瑞他汀藥物之結合物。奧瑞他汀結合微管蛋白已展示干擾微管機構動力學及核及細胞分裂，且具有抗癌活性。通常，基於奧瑞他汀之抗體-藥物結合物包含奧瑞他汀藥物與抗體之間的連接子。奧瑞他汀可在適合與連接子結合之任何位置處鍵聯至抗體。連接子可為例如可裂解連接子(例如肽基連接子)或不可裂解連接子(例如藉由抗體降解釋放之連接子)。奧瑞他汀可為奧瑞斯他汀E或其衍生物。奧瑞他汀可為例如形成於奧瑞他汀E與酮酸之間的酯。例如，可使奧瑞他汀E與對乙醯基苯甲酸或苯甲醯基戊酸反應以分別產生AEB及AEVB。其他典型奧瑞他汀包括MMAF (單甲基奧瑞他汀F)及MMAE (單甲基奧瑞他汀E)。例示性奧瑞他汀之合成及結構描述於美國專利或公開案第7,659,241號、第7,498,298號、第2009-0111756號、第2009-0018086號及第7,968,687號中，其中之每一者以全文引用的方式且出於所有目的併入本文中。Exemplary antibody-drug conjugates include auristatin-based antibody-drug conjugates, ie, conjugates in which the drug component is the auristatin drug. Auristatin binding to tubulin has been shown to interfere with microtubule machinery dynamics and nuclear and cell division, and has anticancer activity. Typically, auristatin-based antibody-drug conjugates comprise a linker between the auristatin drug and the antibody. Auristatin can be linked to the antibody at any position suitable for binding to the linker. The linker can be, for example, a cleavable linker (eg, a peptidyl linker) or a non-cleavable linker (eg, a linker released by antibody degradation). The auristatin can be auristatin E or a derivative thereof. Auristatin can be, for example, an ester formed between auristatin E and a ketoacid. For example, auristatin E can be reacted with p-acetylbenzoic acid or benzylvaleric acid to produce AEB and AEVB, respectively. Other typical auristatins include MMAF (monomethyl auristatin F) and MMAE (monomethyl auristatin E). The synthesis and structure of exemplary auristatins are described in US Patent or Publication No. 7,659,241, No. 7,498,298, No. 2009-0111756, No. 2009-0018086, and No. 7,968,687, each of which is incorporated by reference in its entirety manner and are incorporated herein for all purposes.

例示性基於奧瑞他汀之抗體-藥物結合物包括如下文所展示之vcMMAE、vcMMAF及mcMMAF抗體-藥物結合物，其中Ab為如本文所描述之抗體且val-cit表示纈胺酸-瓜胺酸二肽：

Figure 02_image003
或其醫藥學上可接受之鹽。藥物負載由p (藥物-連接子分子數目/抗體)表示。視上下文而定，p可表示平均藥物-連接子分子數目/抗體，亦稱為平均藥物負載。變量p介於1至20範圍內且較佳為1至8。在一些較佳實施例中，當p表示平均藥物負載時，p在約2至約5範圍內。在一些實施例中，p為約2、約3、約4或約5。在一些態樣中，抗體經由半胱胺酸殘基之硫原子與連接子結合。在一些態樣中，半胱胺酸殘基為經工程改造至抗體中之殘基。在其他態樣中，半胱胺酸殘基為鏈間二硫鍵半胱胺酸殘基。 Exemplary auristatin-based antibody-drug conjugates include vcMMAE, vcMMAF, and mcMMAF antibody-drug conjugates as shown below, wherein Ab is an antibody as described herein and val-cit represents valine-citrulline Dipeptides:
Figure 02_image003
or a pharmaceutically acceptable salt thereof. Drug loading is represented by p (number of drug-linker molecules/antibody). Depending on the context, p may represent the average number of drug-linker molecules per antibody, also known as the average drug load. The variable p is in the range of 1 to 20 and preferably 1 to 8. In some preferred embodiments, p is in the range of about 2 to about 5 when p represents the average drug load. In some embodiments, p is about 2, about 3, about 4, or about 5. In some aspects, the antibody is bound to the linker via the sulfur atom of the cysteine residue. In some aspects, the cysteine residue is a residue engineered into the antibody. In other aspects, the cysteine residue is an interchain disulfide-bonded cysteine residue.

在一些其他實施例中，抗體-藥物結合物具有以全文引用之方式併入本文中之申請案US20160310612A1 (PCT/US2014/060477)中所揭示之連接子單元。在一些其他實施例中，抗體-藥物結合物具有下式(II)：

Figure 02_image005
(II) 其中D為藥物單元，PEG為遮蔽藥物-連接子之疏水性的聚乙二醇單元，L p為允許PEG單元相對於X-D呈並行定向之並聯連結子單元，A為當m大於1，視情況由子單元構成時的分支單元或當m為1時A不存在，X為提供自LDC釋放各D之可釋放組裝單元且Z為L p經由其與抗體L結合之視情況存在之間隔子單元。 In some other embodiments, the antibody-drug conjugate has the linker unit disclosed in application US20160310612A1 (PCT/US2014/060477), which is incorporated by reference in its entirety. In some other embodiments, the antibody-drug conjugate has the following formula (II):
Figure 02_image005
(II) where D is a drug unit, PEG is a polyethylene glycol unit that shields the hydrophobicity of the drug-linker, L p is a parallel linker unit that allows the PEG units to be oriented in parallel with respect to XD, and A is when m is greater than 1 , branching units when optionally composed of subunits or A is absent when m is 1, X is the releasable assembly unit provided to release each D from LDC and Z is the optionally present interval through which L p binds to antibody L subunit.

在一些實施例中，抗體-藥物結合物具有下式III：

Figure 02_image007
(III) 其中AD為使得藉由t指示之X-D部分以平行定向與PEG單元額外連結之藥物連結單元，且L、L p、Z、A、X、D、m、p及s係如式II所定義。 In some embodiments, the antibody-drug conjugate has the following formula III:
Figure 02_image007
(III) wherein AD is a drug linking unit such that the XD moiety indicated by t is additionally linked to the PEG unit in a parallel orientation, and L, Lp , Z, A, X, D, m, p and s are as in Formula II defined.

在其他主要實施例中，本發明之LDC由以下式IV之結構表示：

Figure 02_image009
(IV) 其中AD、L、L p、PEG、Z、A、X、D、m、p、s及t係如式III所定義。 In other main embodiments, the LDC of the present invention is represented by the structure of the following formula IV:
Figure 02_image009
(IV) wherein AD, L, Lp , PEG, Z, A, X, D, m, p, s and t are as defined in formula III.

在一些實施例中，抗體-藥物結合物具有下式1： L-[LU-D'] p( 1) 或其鹽，特定言之醫藥學上可接受之鹽，其中 L為抗體； LU為連接子單元；以及 D'表示式-LU-D'之各藥物連接子部分中之1至藥物單元(D)；以及 下標p為1至12、1至10或1至8之數值或為約4或約8， 其中抗體能夠選擇性結合於腫瘤組織之抗原，以隨後釋放藥物單元作為游離細胞毒性劑， 其中組合物之各抗體-藥物結合物中之式-LU-D'之藥物連接子部分具有式 1A結構：

Figure 02_image011
或其鹽，特定言之醫藥學上可接受之鹽， 其中波浪線指示共價連接至L； D為細胞毒性劑之藥物單元； L B為抗體共價結合部分； A為視情況存在之第一延伸子單元； 下標a為0或1，其分別指示不存在或存在A； B為視情況存在之分支單元； 下標b為0或1，其分別指示不存在或存在B； L O為次要連接子部分，其中次要連接子具有下式：
Figure 02_image013
， 其中與Y相鄰之波浪線指示L O與藥物單元共價連接之位點，且與A'相鄰之波浪線指示共價連接至藥物連接子部分之剩餘部分的位點； A'為視情況存在之第二延伸子單元，其在B不存在的情況下變成A之子單元， 下標a'為0或1，其分別指示不存在或存在A'， W為肽可裂解單元，其中肽可裂解單元為至多12個(例如3至12或3至10個)胺基酸之連續序列，其中序列由選擇性賦予三肽構成，該三肽提供與自比較抗體-藥物結合物組合物之抗體-藥物結合物組合物釋放的細胞毒性劑相比，使腫瘤組織相對於正常組織提高選擇性的暴露於自該組合物之抗體-藥物結合物釋放的細胞毒性劑，在該比較抗體-藥物結合物組合物中，其肽可裂解單元之肽序列為二肽-纈胺酸-瓜胺酸或-纈胺酸-丙胺酸-； 其中腫瘤及正常組織屬於嚙齒動物物種且其中式1組合物提供藉由以下證實之該提高的暴露選擇性： 當以先前針對比較抗體-藥物結合物組合物測定之相同有效量及劑量排程投與時，保持在比較抗體-藥物結合物組合物之腫瘤異種移植模型中之功效，且 展示與比較抗體-藥物結合物組合物之當量(例如相同)投藥相比，當以與腫瘤異種移植模型相同的有效量及劑量排程投與至非腫瘤攜帶嚙齒動物時，自組合物之抗體-藥物結合物釋放的游離細胞毒性劑之血漿濃度降低及/或保存組織中之正常細胞，其中兩種結合物組合物之抗體經未結合抗體置換， 其中對與非腫瘤攜帶嚙齒動物之組織中之正常細胞相同類型之人類組織中之細胞的細胞毒性至少部分地負責經投與治療有效量之比較結合物組合物的人類個體之不良事件； Y為自我分解型間隔子單元；以及 下標y為0、1或2，其分別地指示不存在或存在1或2個Y； 下標p為介於1至4範圍內的整數， 其限制條件為當下標b為0時，下標q為1，且當下標b為1時，下標q為2、3或4；以及 其中組合物之抗體-藥物結合物具有式 1結構，其中下標p經下標p'置換，其中下標p'為1至12、1至10或1至8之整數或為4或8。 In some embodiments, the antibody-drug conjugate has the following formula 1: L-[LU-D'] p ( 1 ) or a salt thereof, in particular a pharmaceutically acceptable salt, wherein L is an antibody; LU is linker unit; and D' represents 1 to drug unit (D) in each drug linker moiety of formula -LU-D'; and the subscript p is a value from 1 to 12, 1 to 10, or 1 to 8 or is about 4 or about 8, wherein the antibody is capable of selectively binding to an antigen of the tumor tissue to subsequently release the drug unit as a free cytotoxic agent, wherein the drug of formula -LU-D' in each antibody-drug conjugate of the composition is linked The subsection has the structure of Formula 1A :
Figure 02_image011
or a salt thereof, specifically a pharmaceutically acceptable salt, wherein the wavy line indicates covalent attachment to L; D is the drug unit of the cytotoxic agent; L B is the covalently bound moiety of the antibody; A is the optional first An extension subunit; the subscript a is 0 or 1, which indicates the absence or presence of A, respectively; B is the branch unit that exists depending on the situation; the subscript b is 0 or 1, which indicates the absence or presence of B, respectively; L O is the secondary linker part, where the secondary linker has the following formula:
Figure 02_image013
, wherein the wavy line adjacent to Y indicates the site where LO is covalently attached to the drug unit, and the wavy line adjacent to A' indicates the site covalently attached to the remainder of the drug linker moiety; A' is an optional second extension subunit that becomes a subunit of A in the absence of B, subscript a' is 0 or 1, which indicates absence or presence of A', respectively, W is a peptide cleavable unit, wherein A peptide cleavable unit is a contiguous sequence of up to 12 (eg, 3 to 12 or 3 to 10) amino acids, wherein the sequence consists of a selectivity-conferring tripeptide that provides an antibody-drug conjugate composition comparable to self Increased selective exposure of tumor tissue relative to normal tissue to a cytotoxic agent released from an antibody-drug conjugate of the composition compared to the cytotoxic agent released from the antibody-drug conjugate composition, where the comparative antibody- In the drug combination composition, the peptide sequence of its peptide cleavable unit is dipeptide-valine-citrulline or -valine-alanine-; wherein tumor and normal tissue belong to rodent species and wherein formula 1 is combined The drug provides this enhanced exposure selectivity as demonstrated by: When administered at the same effective amount and dose schedule previously determined for the comparative antibody-drug conjugate composition, remaining within the range of the comparative antibody-drug conjugate composition Efficacy in tumor xenograft models, and demonstrated compared to equivalent (eg, the same) administration of comparative antibody-drug conjugate compositions, when administered to non-tumor carriers at the same effective amount and dose schedule as in tumor xenograft models In rodents, the plasma concentration of free cytotoxic agents released from the antibody-drug conjugates of the compositions is reduced and/or normal cells in tissues are preserved, wherein the antibodies of the two conjugate compositions are replaced by unconjugated antibodies, wherein the Cytotoxicity of cells in human tissues of the same type as normal cells in tissues of non-tumor bearing rodents is at least partially responsible for adverse events in human subjects administered a therapeutically effective amount of a comparative conjugate composition; Y is self-lysis and the subscript y is 0, 1 or 2, which indicates the absence or the presence of 1 or 2 Y's, respectively; the subscript p is an integer in the range 1 to 4, subject to the subscript When b is 0, subscript q is 1, and when subscript b is 1, subscript q is 2, 3, or 4; and wherein the antibody-drug conjugate of the composition has the structure of formula 1 , wherein subscript p is subscripted by Subscript p' is replaced, where subscript p' is an integer from 1 to 12, 1 to 10, or 1 to 8, or 4 or 8.

一相關實施例提供式 V之藥物連接子： LU'-(D') ( V) 或其鹽，特定言之其醫藥學上可接受之鹽，其中LU'能夠在式 1之L與LU之間提供共價鍵，且因此有時被稱作連接子單元前驅體；且D'表示1至4個藥物單元，其中藥物連接子進一步由式 VI結構定義：

Figure 02_image015
其中L B'能夠轉化為式 VI之L B，由此形成與式 1之L的共價鍵，且因此有時稱為抗體共價結合前驅體部分，且式 VI之剩餘可變基團如關於式 VI所定義。 A related embodiment provides a drug linker of formula V : LU'-(D')( V ) or a salt thereof, in particular a pharmaceutically acceptable salt thereof, wherein LU' can be between L and LU of formula 1 and D' represents 1 to 4 drug units, wherein the drug linker is further defined by the structure of formula VI :
Figure 02_image015
wherein LB ' can be converted to LB of formula VI , thereby forming a covalent bond with L of formula 1 , and is therefore sometimes referred to as an antibody covalently bound precursor moiety, and the remaining variable groups of formula VI are as as defined with respect to Formula VI .

在一些實施例中，ADC包含與mc-vc-PABC-MMAE (在本文中亦稱作vcMMAE或1006)、mc-vc-PABC-MMAF、mc-MMAF或mp-dLAE-PABC-MMAE (在本文中亦稱作dLAE-MMAE、mp-dLAE-MMAE或7092)結合之抗體(例如如本文所描述之抗體)或其醫藥學上可接受之鹽。mp-dLAE-PABC-MMAE描述於PCT公開案第WO 2021/055865 A1號中。此類ADC展示如下，其中Ab包含抗原結合蛋白(例如如本文所描述之任何抗體)，mc表示順丁烯二醯亞胺基己醯基，mp係指順丁烯二醯亞胺丙醯基：

Figure 02_image017
， val-cit (vc)表示纈胺酸-瓜胺酸二肽，PABC表示對胺基苯甲氧基羰基且dLAE表示D-白胺酸-丙胺酸-麩胺酸三肽：
Figure 02_image019
mp-dLAE-PABC-MMAE。在一些實施例中，藥物負載由p(藥物-連接子分子之數目/抗體)表示。在一些實施例中，p可表示抗體組合物中之平均藥物-連接子分子數目/抗體，亦稱為平均藥物負載。在一些實施例中，p介於1至20範圍內。在一些實施例中，p介於1至8範圍內。在一些實施例中，當p表示平均藥物負載時，p介於約2至約5範圍內。在一些實施例中，p為約2、約3、約4或約5。在一些實施例中，製劑中之平均藥物數目/抗體可藉由諸如質譜分析、HIC、ELISA分析及HPLC之習知手段來表徵。在一些實施例中，抗原結合蛋白(例如抗體)經由抗體之半胱胺酸殘基連接至藥物-連接子。在一些實施例中，半胱胺酸殘基為經工程改造至抗體中之殘基。在一些實施例中，半胱胺酸殘基為鏈間二硫鍵半胱胺酸殘基。 C. 例示性 ADC In some embodiments, the ADC comprises a combination with mc-vc-PABC-MMAE (also referred to herein as vcMMAE or 1006), mc-vc-PABC-MMAF, mc-MMAF, or mp-dLAE-PABC-MMAE (herein (also known as dLAE-MMAE, mp-dLAE-MMAE, or 7092)-binding antibody (eg, an antibody as described herein) or a pharmaceutically acceptable salt thereof. mp-dLAE-PABC-MMAE is described in PCT Publication No. WO 2021/055865 A1. Such ADCs are shown below, wherein Ab comprises an antigen binding protein (eg, any antibody as described herein), mc represents maleimidohexanoyl, and mp refers to maleimidopropionyl :
Figure 02_image017
, val-cit (vc) denotes valine-citrulline dipeptide, PABC denotes p-aminobenzyloxycarbonyl and dLAE denotes D-leucine-alanine-glutamic acid tripeptide:
Figure 02_image019
mp-dLAE-PABC-MMAE. In some embodiments, drug loading is represented by p(number of drug-linker molecules/antibody). In some embodiments, p may represent the average number of drug-linker molecules per antibody in the antibody composition, also referred to as the average drug load. In some embodiments, p ranges from 1 to 20. In some embodiments, p ranges from 1 to 8. In some embodiments, p ranges from about 2 to about 5 when p represents the average drug load. In some embodiments, p is about 2, about 3, about 4, or about 5. In some embodiments, the average number of drug/antibody in the formulation can be characterized by conventional means such as mass spectrometry, HIC, ELISA analysis, and HPLC. In some embodiments, the antigen binding protein (eg, antibody) is linked to the drug-linker via a cysteine residue of the antibody. In some embodiments, the cysteine residue is a residue engineered into the antibody. In some embodiments, the cysteine residue is an interchain disulfide cysteine residue. C. Exemplary ADCs

用於本發明方法之非限制性例示性ADC包括ADC，其包含結合本文中所論述的任何例示性目標之抗體，該抗體與本文所描述之任何微管蛋白干擾劑結合。Non-limiting exemplary ADCs for use in the methods of the invention include ADCs comprising antibodies that bind any of the exemplary targets discussed herein that bind to any of the tubulin interfering agents described herein.

在一些實施例中，ADC為抗唾液酸基Tn抗原抗體-ADC，其包含結合於唾液酸基Tn抗原(sTn)及MMAE之抗體。 參見例如美國專利公開案第2018/0327509A1號；第WO2017083582A1號；本文中之序列表。 In some embodiments, the ADC is an anti-sialyl-Tn antigen antibody-ADC comprising antibodies that bind to a sialyl-Tn antigen (sTn) and MMAE. See, eg , US Patent Publication No. 2018/0327509A1; No. WO2017083582A1; Sequence Listing herein.

在一些實施例中，ADC為貝蘭單抗莫福汀，其包含結合於B細胞成熟抗原(BCMA)及MMAF之抗體。 參見例如美國專利第9,273,141號。 In some embodiments, the ADC is belantezumab mofotine, which comprises antibodies that bind to B cell maturation antigen (BCMA) and MMAF. See, eg , US Patent No. 9,273,141.

在一些實施例中，ADC為抗密連蛋白-18.2 ADC，其包含奧瑞他汀及如下抗體： 唑貝妥西單抗(175D10)，其揭示於美國專利案第8,168,427號中且包含：包含SEQ ID NO:59之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:60之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:61至66之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3； 163E12，其揭示於美國專利案第8,168,427號中且包含包含SEQ ID NO:67之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO: 68之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:69至74之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3； 任何抗密連蛋白-18.2抗體，其揭示於PCT公開案第WO 2020/135674 A1號中；或 任何抗密連蛋白-18.2抗體，其揭示於PCT公開案第WO 2021/032157 A1號中。 In some embodiments, the ADC is an anti-claudin-18.2 ADC comprising auristatin and the following antibodies: Zobetuzumab (175D10), disclosed in US Pat. No. 8,168,427 and comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:59 and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:60 a light chain variable region (VL) of an amino acid sequence; or a heavy chain CDR1, CDR2 and CDR3 and a light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 61 to 66, respectively; 163E12, disclosed in US Pat. No. 8,168,427 and comprising a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 67 and a light chain comprising the amino acid sequence of SEQ ID NO: 68 can be A variable region (VL); or comprising heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 69 to 74, respectively; any anti-claudin-18.2 antibody disclosed in PCT Publication No. WO 2020/135674 A1; or Any of the anti-Claudin-18.2 antibodies disclosed in PCT Publication No. WO 2021/032157 A1.

在一些實施例中，ADC為SGN-PDL1V，其包含抗PD-L1抗體及MMAE，該抗體包含：包含SEQ ID NO:75之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:76之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:77至82之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。In some embodiments, the ADC is SGN-PDL1V comprising an anti-PD-L1 antibody and MMAE comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:75 and comprising SEQ ID NO: 75 The light chain variable region (VL) of the amino acid sequence of NO: 76; or the heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3 comprising the amino acid sequence of SEQ ID NO: 77 to 82, respectively .

在一些實施例中，ADC為SGN-ALPV，其包含抗ALP抗體及MMAE，該抗體包含：包含SEQ ID NO:83之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:84之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:85至90之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。In some embodiments, the ADC is SGN-ALPV comprising an anti-ALP antibody and MMAE comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 83 and comprising SEQ ID NO: The light chain variable region (VL) of the amino acid sequence of 84; or the heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 85 to 90, respectively.

在一些實施例中，ADC為SGN-B7H4V，其包含抗B7H4抗體及MMAE，該抗體包含：包含SEQ ID NO:91之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:92之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:93至98之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。In some embodiments, the ADC is SGN-B7H4V comprising an anti-B7H4 antibody and MMAE comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:91 and comprising: The light chain variable region (VL) of the amino acid sequence of 92; or the heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 93 to 98, respectively.

在一些實施例中，ADC為迪西妥單抗維多汀，其包含抗HER2抗體及MMAE，該抗體包含：包含SEQ ID NO:99之胺基酸序列的重鏈及包含SEQ ID NO:100之胺基酸序列的輕鏈。In some embodiments, the ADC is dicituzumab vedotin, comprising an anti-HER2 antibody and MMAE, the antibody comprising: a heavy chain comprising the amino acid sequence of SEQ ID NO:99 and comprising SEQ ID NO:100 the amino acid sequence of the light chain.

在一些實施例中，ADC為立伐土珠單抗維多汀，其包含抗NaPi2B抗體及MMAE，該抗體包含：包含SEQ ID NO:101之胺基酸序列的重鏈及包含SEQ ID NO:102之胺基酸序列的輕鏈。In some embodiments, the ADC is rivartuzumab vedotin comprising an anti-NaPi2B antibody and MMAE, the antibody comprising: a heavy chain comprising the amino acid sequence of SEQ ID NO: 101 and comprising SEQ ID NO: The light chain of the amino acid sequence of 102.

在一些實施例中，ADC為恩諾單抗維多汀，其包含結合連接素-4之抗體及MMAE。 參見例如美國專利第8,637,642號；WO 2012/047724。在一些實施例中，恩諾單抗維多汀之抗體包含：包含SEQ ID NO:103之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:104之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:105至110之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。 In some embodiments, the ADC is ennozumab vedotin, which comprises an antibody that binds connexin-4 and MMAE. See, eg , US Patent No. 8,637,642; WO 2012/047724. In some embodiments, the antibody to ennozumab vedotin comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 103 and a variable region (VH) comprising the amino acid sequence of SEQ ID NO: 104 light chain variable region (VL); or comprising heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 105 to 110, respectively.

在一些實施例中，ADC為SGN-B6A，其包含結合於AVB6之抗體及MMAE。在一些實施例中，SGN-B6A包含：包含SEQ ID NO:37之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:38之胺基酸序列的輕鏈可變區(VL)。在一些實施例中，ADC包含抗AVB6抗體，該抗體包含：包含SEQ ID NO:111之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:112之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:113至118之胺基酸序列重鏈CDR1、CDR2、CDR3以及輕鏈CDR1、CDR2及CDR3。在一些實施例中，ADC包含抗AVB6抗體，該抗體包含包含SEQ ID NO:119之胺基酸序列重鏈可變區(VH)及包含SEQ ID NO:120之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:121至126之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。In some embodiments, the ADC is SGN-B6A, which comprises an antibody that binds to AVB6 and MMAE. In some embodiments, SGN-B6A comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:37 and a light chain variable region comprising the amino acid sequence of SEQ ID NO:38 ( VL). In some embodiments, the ADC comprises an anti-AVB6 antibody comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 111 and a light chain comprising the amino acid sequence of SEQ ID NO: 112 chain variable region (VL); or comprising the amino acid sequences comprising SEQ ID NOs: 113 to 118, respectively, heavy chain CDR1, CDR2, CDR3 and light chain CDR1, CDR2 and CDR3. In some embodiments, the ADC comprises an anti-AVB6 antibody comprising a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 119 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 120 variable region (VL); or comprising heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 121 to 126, respectively.

在一些實施例中，ADC為抗CD228抗體-ADC，其包含結合於CD228之抗體及MMAE。 參見例如美國專利公開案第2020/0246479A1號；WO2020/163225A1。在一些實施例中，ADC為SGN-CD228A，其包含抗CD228抗體及MMAE，該抗體包含：包含SEQ ID NO:127之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:128之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:129至134之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。 In some embodiments, the ADC is an anti-CD228 antibody-ADC comprising an antibody that binds to CD228 and MMAE. See, eg , US Patent Publication No. 2020/0246479A1; WO2020/163225A1. In some embodiments, the ADC is SGN-CD228A comprising an anti-CD228 antibody and MMAE comprising: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 127 and comprising SEQ ID NO: The light chain variable region (VL) of the amino acid sequence of 128; or the heavy chain CDR1, CDR2 and CDR3 and the light chain CDR1, CDR2 and CDR3 comprising the amino acid sequence of SEQ ID NOs: 129 to 134, respectively.

在一些實施例中，ADC為SGN-LIV1A (拉地妥珠單抗維多汀；LV)，其包含抗LIV-1抗體及MMAE，該抗體包含包含SEQ ID NO:135之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:136之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:137至142之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3； 其中SGN-LIV1A包含與以下結合之抗LIV-1抗體：mc-vc-PABC-MMAE、mc-vc-PABC-MMAF、mc-MMAF或mp-dLAE-PABC-MMAE。 In some embodiments, the ADC is SGN-LIV1A (latetuzumab vedotin; LV), which comprises an anti-LIV-1 antibody, and MMAE, which comprises an antibody comprising the amino acid sequence of SEQ ID NO: 135 A heavy chain variable region (VH) and a light chain variable region (VL) comprising the amino acid sequence of SEQ ID NO: 136; or a heavy chain CDR1 comprising the amino acid sequence of SEQ ID NO: 137 to 142, respectively , CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3; wherein SGN-LIV1A comprises an anti-LIV-1 antibody conjugated to: mc-vc-PABC-MMAE, mc-vc-PABC-MMAF, mc-MMAF or mp-dLAE-PABC-MMAE.

在一些實施例中，ADC為替索圖單抗維多汀(TV)，其包含結合組織因子(TF)之抗體及MMAE。 參見例如美國專利第9,168,314號及第9,150,658號；WO 2011/157741；WO 2010/066803。在一些實施例中，TV之抗體包含：包含SEQ ID NO:143之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:144之胺基酸序列的輕鏈可變區(VL)；或包含分別包含SEQ ID NO:145至150之胺基酸序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3。 In some embodiments, the ADC is tesotumumab vedotin (TV), which comprises an antibody that binds tissue factor (TF) and MMAE. See, eg , US Patent Nos. 9,168,314 and 9,150,658; WO 2011/157741; WO 2010/066803. In some embodiments, the antibody to TV comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 143 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 144 ( VL); or comprising heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 145 to 150, respectively.

在一些實施例中，ADC包含MMAE且結合選自以下之目標：AMHRII、Axl、CA9、CD142、CD20、CD22、CD228、CD248、CD30、CD33、CD7、CD48、CD71、CD79b、CLDN18.2、CLDN6、c-MET、EGFR、EphA2、ETBR、FCRH5、GCC、Globo H、gpNMB、HER-2、IL7R、整合素β-6、KAAG-1、LGR5、LIV-1、LRRC15、Ly6E、間皮素(MSLN)、MET、MRC2、MUC16、NaPi2b、連接素-4、OT-MUC1 (腫瘤栓繫MUC1)、PSMA、ROR1、SLAMF7、SLC44A4、SLITRK6、STEAP-1、STn、TIM-1、TRA-1-60、腫瘤相關糖蛋白72 (TAG-72)。In some embodiments, the ADC comprises MMAE and binds a target selected from AMHRII, Axl, CA9, CD142, CD20, CD22, CD228, CD248, CD30, CD33, CD7, CD48, CD71, CD79b, CLDN18.2, CLDN6 , c-MET, EGFR, EphA2, ETBR, FCRH5, GCC, Globo H, gpNMB, HER-2, IL7R, integrin beta-6, KAAG-1, LGR5, LIV-1, LRRC15, Ly6E, mesothelin ( MSLN), MET, MRC2, MUC16, NaPi2b, connexin-4, OT-MUC1 (tumor tethered MUC1), PSMA, ROR1, SLAMF7, SLC44A4, SLITRK6, STEAP-1, STn, TIM-1, TRA-1- 60. Tumor-associated glycoprotein 72 (TAG-72).

在一些實施例中，ADC包含MMAE且為以下中之一者：DP303c，亦稱為SYSA1501，其靶向HER-2 (CSPC Pharmaceutical；Dophen Biomed)；SIA01-ADC，亦稱為ST1，其靶向STn (Siamab Therapeutics)；拉地妥珠單抗維多汀(Ladiratuzumab vedotin)，亦稱為SGN-LIV1A，其靶向LIV-1 (Merck & Co., Inc.；Seagen (Seattle Genetics) Inc.)；ABBV-085，亦稱為沙馬妥單抗維多汀(Samrotamab vedotin)，其靶向LRRC15 (Abbvie; Seagen (Seattle Genetics) Inc.)；DMOT4039A，亦稱為RG7600；αMSLN-MMAE，其靶向間皮素(MSLN) (Roche-Genentech)；RC68，亦稱為Remegen EGFR ADC，其靶向EGFR (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.))；RC108，亦稱為RC108-ADC，其靶向c-MET (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.))；CMG901，亦稱為MRG005，其靶向CLDN18.2 (Keymed Biosciences；Lepu biotech；Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd))；YBL-001，亦稱為LCB67，其靶向DLK-1 (Lego Chem Biosciences；Pyxis Oncology；Y-Biologics)；DCDS0780A，亦稱為艾妥珠單抗維多汀(Iladatuzumab vedotin)；RG7986，其靶向CD79b (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；替索圖單抗維多汀(Tisotumab vedotin)，亦稱為Humax-TF-ADC；tf-011-mmae；TIVDAK™，其靶向CD142 (GenMab; Seagen (Seattle Genetics) Inc.)；GO-3D1-ADC，亦稱為humAb-3D1-MMAE ADC，其靶向MUC1-C (Genus Oncology LLC)；ALT-P7，亦稱為HM2-MMAE，其靶向HER-2 (Alteogen, Inc.；Levena Biopharma；3SBio, Inc.)；凡多土珠單抗維多汀(Vandortuzumab vedotin)，亦稱為DSTP3086S；RG7450，其靶向STEAP-1 (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；立伐土珠單抗維多汀(Lifastuzumab Vedotin)，亦稱為DNIB0600A；NaPi2b ADC；RG7599，其靶向NaPi2b (Roche-Genentech)；索菲土珠單抗維多汀(Sofituzumab vedotin)，亦稱為DMUC5754A；RG7458，其靶向MUC16 (Seagen (Seattle Genetics) Inc.；Roche-Genentech)；RG7841，亦稱為DLYE5953A，其靶向Ly6E (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；RG7598，亦稱為DFRF4539A，其靶向FCRH5 (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；RG7636，亦稱為DEDN6526A，其靶向ETBR (Seagen (Seattle Genetics) Inc.；Roche-Genentech)；匹納土珠單抗維多汀，亦稱為DCDT2980S；RG7593，其靶向CD22 (Roche-Genentech)；波妥珠單抗維多汀，亦稱為DCDS4501A；POLIVY™；RG7596；RO-5541077，其靶向CD79b (Chugai Pharmaceutical；Roche-Genentech；Seagen (Seattle Genetics) Inc.)；DMUC4064A，亦稱為D-4064a；RG7882，其靶向MUC16 (Roche-Genentech；Seagen (Seattle Genetics) Inc.)；SYSA1801，亦稱為CPO102，其靶向CLDN18.2 (Conjupro Biotherapeutics Inc.；CSPC ZhongQi Pharmaceutical Technology Co.)；RC118，亦稱為密連蛋白18.2-ADC；YH005，其靶向CLDN18.2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)；Biocytogen)；VLS-101，亦稱為瑟吐珠單抗維多汀；MK-2140；UC-961ADC3；澤羅妥單抗維多汀(Zilovertamab Vedotin)，其靶向ROR1 (VelosBio. Inc)；格雷巴單抗維多汀(Glembatumumab vedotin)，亦稱為CDX-011；CR011-vcMMAE，其靶向gpNMB (Celldex Therapeutics)；BA3021，亦稱為CAB-ROR2-ADC；奧珠單抗維多汀(Ozuriftamab Vedotin)，其靶向ROR2 (Bioatla；Himalaya Therapeutics)；BA3011，亦稱為CAB-AXL-ADC；美克波單抗維多汀(Mecbotamab Vedotin)，其靶向Axl (Bioatla; Himalaya Therapeutics)；CM-09，亦稱為Bstrongximab-ADC，其靶向TRA-1-60 (CureMeta)；ABBV-838，亦稱為阿妥昔珠單抗維多汀(Azintuxizumab vedotin)，其靶向SLAMF7 (Abbvie)；依那波他單抗維多汀(Enapotamab vedotin)，亦稱為AXL-107-MMAE；HuMax-AXL-ADC，其靶向Axl (GenMab; Seagen (Seattle Genetics) Inc.)；ARC-01，亦稱為抗CD79b ADC，其靶向CD79b (Araris Biotech AG)；迪西妥單抗維多汀(Disitamab vedotin)，亦稱為Aidexi®；RC48，其靶向HER-2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)；Seagen (Seattle Genetics) Inc.)；ASG-5ME，亦稱為AGS-5；AGS-5ME，其靶向SLC44A4 (Agensys, Inc.；Astellas Pharma Inc.；Seagen (Seattle Genetics) Inc.)；恩諾單抗維多汀(Enfortumab vedotin)，亦稱為AGS-22M6E；ASG-22CE；ASG-22ME；PADCEV™，其靶向連接素-4 (Astellas Pharma Inc.；Seagen (Seattle Genetics) Inc.)；ASG-15ME，亦稱為AGS-15E；斯妥尤單抗維多汀(Sirtratumab vedotin)，其靶向SLITRK6 (Seagen (Seattle Genetics) Inc.；Astellas Pharma Inc.)；本妥昔單抗維多汀(Brentuximab vedotin)，亦稱為Adcetris；cAC10-vcMMAE；SGN-35，其靶向CD30 (Seagen (Seattle Genetics) Inc.；Takeda)；肽利索單抗維多汀(Telisotuzumab vedotin)，亦稱為ABBV-399，其靶向c-MET (Abbvie)；羅妥昔珠單抗維多汀(Losatuxizumab vedotin)，亦稱為ABBV-221，其靶向EGFR (Abbvie)；CX-2029，亦稱為ABBV-2029，其靶向CD71 (Abbvie；CytomX Therapeutics)；AB-3A4-ADC，亦稱為AB-3A4-vcMMAE，其靶向KAAG-1 (Alethia Biotherapeutics)；因杜薩土單抗維多汀(Indusatumab vedotin)，亦稱為5F9-vcMMAE；MLN0264；TAK-264，其靶向GCC (Takeda；Millennium Pharmaceuticals, Inc)；FOR46，其靶向CD46 (Fortis Therapeutics, Inc.)；LR004-VC-MMAE，其靶向EGFR (Chinese Academy of Medical Sciences Peking Union Medical College Hospital)；CD30-ADC，其靶向CD30 (NBE Therapeutics；Boehringer Ingelheim)；抗內皮唾酸蛋白-MC-VC-PABC-MMAE，其靶向CD248 (Genzyme)；OBI-998，其靶向SSEA-4 (OBI Pharma)；MRG002，其靶向HER-2 (Lepu biotech；Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd))；TRS005，其靶向CD20 (Teruisi Pharmaceuticals)；Oba01，其靶向DR5 (死亡受體5) (Obio Technology (Shanghai) Corp.,Ltd.；Yantai Obioadc Biomedical Technology Ltd.)；PSMA ADC，其靶向PSMA (Progenics Pharmaceuticals, Inc；Seagen (Seattle Genetics) Inc.)；SGN-CD48A，其靶向CD48 (Seagen (Seattle Genetics) Inc.)；IMAB362-vcMMAE，其靶向CLDN18.2 (Astellas Pharma Inc.; Ganymed)；GB251，其靶向HER-2 (Genor Biopharma Co., Ltd.)；Innate Pharma BTG-ADC，其靶向CD30 (Innate Pharma; Sanofi)；ADCendo uPARAP ADC，其靶向MRC2 (ADCendo)；XCN-010，其靶向actM (Xiconic Pharmaceuticals, LLC)；ANT-043，其靶向HER-2 (Antikor Biopharma)；OBI-999，其靶向Globo H (Abzena; OBI Pharma)；LY3343544，其靶向MET (Eli Lilly and Company)；Tagworks抗TAG72 ADC，其靶向TAG-72 (Tagworks Pharmaceuticals)；IMAB027-vcMMAE，其靶向CLDN6 (Ganymed; Astellas Pharma Inc.)；LGR5-ADC，其靶向LGR5 (Genentech, Inc.)；Philochem B12-MMAE ADC，其靶向IL-7R (Instituto de Medicina Molecular João Lobo Antunes; Philochem AG)；TE-1522，其靶向CD19 (Immunwork)；SGN-STNV，其靶向STn (Seagen (Seattle Genetics) Inc.)；HTI-1511，其靶向EGFR (Abzena; Halozyme Therapeutics)；Peptron PAb001-ADC，其靶向OT-MUC1 (腫瘤栓繫-MUC1) (Peptron; Qilu Pharmaceutical co. Ltd.)；LM-102，其靶向CLDN18.2 (LaNova Medicines Limited)；Anwita Biosciences MSLN-MMAE，其靶向間皮素(MSLN) (Anwita biosciences)；SGN-CD228A，其靶向CD228 (Seagen (Seattle Genetics) Inc.)；NBT828，其靶向HER-2 (NewBio Therapeutics; Genor Biopharma Co., Ltd.)；Gamamabs GM103，其靶向AMHR2 (GamaMabs Pharma; Exelixis)；LCB14-0302，其靶向HER-2 (Lego Chem Biosciences)；BAY79-4620，其靶向碳酸酐酶IX (CAIX) (Bayer; MorphoSys)；NBT508，其靶向CD79b (NewBio Therapeutics)；PAT-DX3-MMAE，其靶向未揭示(Patrys; Yale University)；AGS67E，其靶向CD37 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.)；CDX-014，其靶向TIM-1 (Celldex Therapeutics)；BVX001，其靶向CD33；CD7 (Bivictrix therapeutics)；SGN-B6A，其靶向整合素β-6 (Seagen (Seattle Genetics) Inc.)；MRG003，其靶向EGFR (Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd))及PYX-202，其靶向DLK-1 (Pyxis Oncology; Lego Chem Biosciences)。In some embodiments, the ADC comprises MMAE and is one of: DP303c, also known as SYSA1501, which targets HER-2 (CSPC Pharmaceutical; Dophen Biomed); SIA01-ADC, also known as ST1, which targets STn (Siamab Therapeutics); Ladiratuzumab vedotin, also known as SGN-LIV1A, which targets LIV-1 (Merck & Co., Inc.; Seagen (Seattle Genetics) Inc.) ; ABBV-085, also known as Samrotamab vedotin, which targets LRRC15 (Abbvie; Seagen (Seattle Genetics) Inc.); DMOT4039A, also known as RG7600; αMSLN-MMAE, which targets To mesothelin (MSLN) (Roche-Genentech); RC68, also known as Remegen EGFR ADC, which targets EGFR (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)); RC108, also known as RC108-ADC , which targets c-MET (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.)); CMG901, also known as MRG005, which targets CLDN18.2 (Keymed Biosciences; Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya) Biotechnology Co., Ltd)); YBL-001, also known as LCB67, which targets DLK-1 (Lego Chem Biosciences; Pyxis Oncology; Y-Biologics); DCDS0780A, also known as Atuzumab vedotin ( Iladatuzumab vedotin); RG7986, which targets CD79b (Roche-Genentech; Seagen (Seattle Genetics) Inc.); Tisotumab vedotin, also known as Humax-TF-ADC; tf-011- mmae; TIVDAK™, which targets CD142 (GenMab; Seagen (Seattle Genetics) Inc. ); GO-3D1-ADC, also known as humAb-3D1-MMAE ADC, which targets MUC1-C (Genus Oncology LLC); ALT-P7, also known as HM2-MMAE, which targets HER-2 (Alteogen, Inc.; Levena Biopharma; 3SBio, Inc.); Vandortuzumab vedotin, also known as DSTP3086S; RG7450, which targets STEAP-1 (Roche-Genentech; Seagen (Seattle Genetics) Inc. .); Lifastuzumab Vedotin, also known as DNIB0600A; NaPi2b ADC; RG7599, which targets NaPi2b (Roche-Genentech); Sofituzumab vedotin , also known as DMUC5754A; RG7458, which targets MUC16 (Seagen (Seattle Genetics) Inc.; Roche-Genentech); RG7841, also known as DLYE5953A, which targets Ly6E (Roche-Genentech; Seagen (Seattle Genetics) Inc.) RG7598, also known as DFRF4539A, which targets FCRH5 (Roche-Genentech; Seagen (Seattle Genetics) Inc.); RG7636, also known as DEDN6526A, which targets ETBR (Seagen (Seattle Genetics) Inc.; Roche-Genentech) ; Pinatuzumab vedotin, also known as DCDT2980S; RG7593, which targets CD22 (Roche-Genentech); Perstuzumab vedotin, also known as DCDS4501A; POLIVY™; RG7596; RO-5541077 , which targets CD79b (Chugai Pharmaceutical; Roche-Genentech; Seagen (Seattle Genetics) Inc.); DMUC4064A, also known as D-4064a; RG7882, which targets MUC16 (Roche-Genentech; Seagen (Seattle Genetics) Inc.) ; SYSA1801, also known as CPO102, targets CLDN18.2 (Conjupro Biot herapeutics Inc.; CSPC ZhongQi Pharmaceutical Technology Co.); RC118, also known as claudin 18.2-ADC; YH005, which targets CLDN18.2 (RemeGen (Rongchang Biopharmaceutical (Yantai) Co., Ltd.); Biocytogen); VLS-101, also known as cetozumab vedotin; MK-2140; UC-961ADC3; Zilvertamab Vedotin, which targets ROR1 (VelosBio. Inc); Anti-vedotin (Glembatumumab vedotin), also known as CDX-011; CR011-vcMMAE, which targets gpNMB (Celldex Therapeutics); BA3021, also known as CAB-ROR2-ADC; Ozuriftamab Vedotin ), which targets ROR2 (Bioatla; Himalaya Therapeutics); BA3011, also known as CAB-AXL-ADC; Mecbotamab Vedotin, which targets Axl (Bioatla; Himalaya Therapeutics); CM- 09, also known as Bstrongximab-ADC, which targets TRA-1-60 (CureMeta); ABBV-838, also known as Azintuxizumab vedotin, which targets SLAMF7 (Abbvie); Enapotamab vedotin, also known as AXL-107-MMAE; HuMax-AXL-ADC, which targets Axl (GenMab; Seagen (Seattle Genetics) Inc.); ARC-01, also known as is an anti-CD79b ADC, which targets CD79b (Araris Biotech AG); Disitamab vedotin, also known as Aidexi®; RC48, which targets HER-2 (RemeGen (Rongchang Biopharmaceutical (Yantai)) Co., Ltd.); Seagen (Seattle Genetics) Inc.); ASG-5ME, also known as AGS-5; AGS-5ME, which targets SLC44A4 (Agensys, Inc. Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.); Enfortumab vedotin, also known as AGS-22M6E; ASG-22CE; ASG-22ME; PADCEV™, which targets connexin -4 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.); ASG-15ME, also known as AGS-15E; Sirtratumab vedotin, which targets SLITRK6 (Seagen (Seattle Genetics) Inc.) ) Inc.; Astellas Pharma Inc.); Brentuximab vedotin, also known as Adcetris; cAC10-vcMMAE; SGN-35, which targets CD30 (Seagen (Seattle Genetics) Inc.; Takeda ); Telisotuzumab vedotin, also known as ABBV-399, which targets c-MET (Abbvie); Losatuxizumab vedotin, also known as ABBV- 221, which targets EGFR (Abbvie); CX-2029, also known as ABBV-2029, which targets CD71 (Abbvie; CytomX Therapeutics); AB-3A4-ADC, also known as AB-3A4-vcMMAE, which targets KAAG-1 (Alethia Biotherapeutics); Indusatumab vedotin, also known as 5F9-vcMMAE; MLN0264; TAK-264, which targets GCC (Takeda; Millennium Pharmaceuticals, Inc); FOR46, It targets CD46 (Fortis Therapeutics, Inc.); LR004-VC-MMAE, which targets EGFR (Chinese Academy of Medical Sciences Peking Union Medical College Hospital); CD30-ADC, which targets CD30 (NBE Therapeutics; Boehringer Ingelheim) ; anti-endosialin-MC-VC-PABC-MMAE, which targets CD248 (Genzy me); OBI-998, which targets SSEA-4 (OBI Pharma); MRG002, which targets HER-2 (Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd)); TRS005, which targets CD20 (Teruisi Pharmaceuticals); Oba01, which targets DR5 (Death Receptor 5) (Obio Technology (Shanghai) Corp., Ltd.; Yantai Obioadc Biomedical Technology Ltd.); PSMA ADC, which targets PSMA (Progenics Pharmaceuticals, Inc Seagen (Seattle Genetics) Inc.); SGN-CD48A, which targets CD48 (Seagen (Seattle Genetics) Inc.); IMAB362-vcMMAE, which targets CLDN18.2 (Astellas Pharma Inc.; Ganymed); GB251, which targets CLDN18.2 (Astellas Pharma Inc.; Ganymed); Targets HER-2 (Genor Biopharma Co., Ltd.); Innate Pharma BTG-ADC, which targets CD30 (Innate Pharma; Sanofi); ADCendo uPARAP ADC, which targets MRC2 (ADCendo); XCN-010, which targets To actM (Xiconic Pharmaceuticals, LLC); ANT-043, which targets HER-2 (Antikor Biopharma); OBI-999, which targets Globo H (Abzena; OBI Pharma); LY3343544, which targets MET (Eli Lilly and Company); Tagworks anti-TAG72 ADC, which targets TAG-72 (Tagworks Pharmaceuticals); IMAB027-vcMMAE, which targets CLDN6 (Ganymed; Astellas Pharma Inc.); LGR5-ADC, which targets LGR5 (Genentech, Inc.) ; Philochem B12-MMAE ADC, which targets IL-7R (Instituto de Medicina Molecular João Lobo Antunes; Philo chem AG); TE-1522, which targets CD19 (Immunwork); SGN-STNV, which targets STn (Seagen (Seattle Genetics) Inc.); HTI-1511, which targets EGFR (Abzena; Halozyme Therapeutics); Peptron PAb001-ADC, which targets OT-MUC1 (Tumor Tethered-MUC1) (Peptron; Qilu Pharmaceutical co. Ltd.); LM-102, which targets CLDN18.2 (LaNova Medicines Limited); Anwita Biosciences MSLN-MMAE, It targets mesothelin (MSLN) (Anwita biosciences); SGN-CD228A, which targets CD228 (Seagen (Seattle Genetics) Inc.); NBT828, which targets HER-2 (NewBio Therapeutics; Genor Biopharma Co., Ltd .); Gamamabs GM103, which targets AMHR2 (GamaMabs Pharma; Exelixis); LCB14-0302, which targets HER-2 (Lego Chem Biosciences); BAY79-4620, which targets carbonic anhydrase IX (CAIX) (Bayer; MorphoSys); NBT508, which targets CD79b (NewBio Therapeutics); PAT-DX3-MMAE, which targets undisclosed (Patrys; Yale University); AGS67E, which targets CD37 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc. .); CDX-014, which targets TIM-1 (Celldex Therapeutics); BVX001, which targets CD33; CD7 (Bivictrix therapeutics); SGN-B6A, which targets integrin beta-6 (Seagen (Seattle Genetics) Inc .); MRG003, which targets EGFR (Lepu biotech; Shanghai Miracogen Inc. (Shanghai Meiya Biotechnology Co., Ltd)) and PYX-202, which targets DLK-1 (Pyxis Oncolo gy; Lego Chem Biosciences).

在一些實施例中，ADC包含MMAF且結合選自以下之目標：BCMA、GPC-1、CD30、c-MET、SAIL、HER-3、CD70、CD46、HER-2、5T4、ENPP3、CD19、EGFR、EphA2。In some embodiments, the ADC comprises MMAF and binds a target selected from BCMA, GPC-1, CD30, c-MET, SAIL, HER-3, CD70, CD46, HER-2, 5T4, ENPP3, CD19, EGFR , EphA2.

在一些實施例中，ADC包含MMAF且為以下中之一者：CD70-ADC，其靶向CD70 (Kochi University; Osaka University)；IGN786，其靶向SAIL (AstraZeneca；Igenica Biotherapeutics)；PF-06263507，其靶向5T4 (Pfizer)；GPC1-ADC，其靶向GPC-1 (Kochi University)；ADC-AVP10，其靶向CD30 (Avipep)；M290-MC-MMAF，其靶向CD103 (The Second Affiliated Hospital of Harbin Medical University)；BVX001，其靶向CD33；CD7 (Bivictrix therapeutics)；Tanabe P3D12-vc-MMAF，其靶向c-MET (Tanabe Research Laboratories)；LILRB4-靶向ADC，其靶向LILRB4 (The University of Texas Health Science Center, Houston)；TSD101，亦稱為ABL201，其靶向BCMA (TSD Life Science；ABL Bio；Lego Chem Biosciences)；瑪汀-迪妥昔珠單抗莫福汀，亦稱為ABT-414，其靶向EGFR (Abbvie; Seagen (Seattle Genetics) Inc.)；AGS16F，亦稱為AGS-16C3F；AGS-16M8F，其靶向ENPP3 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.)；AVG-A11 BCMA ADC，亦稱為AVG-A11-mcMMAF，其靶向BCMA (Avantgen)；貝蘭他單抗莫福汀，亦稱為BLENREP；GSK2857916；J6M0-mcMMAF，其靶向BCMA (GlaxoSmithKline；Seagen (Seattle Genetics) Inc.)；MP-HER3-ADC，亦稱為HER3-ADC，其靶向HER-3 (MediaPharma)；FS-1502，亦稱為LCB14-0110，其靶向HER-2 (Lego Chem Biosciences;Shanghai Fosun Pharmaceutical Development Co, Ltd.)；MEDI-547，亦稱為MI-CP177，其靶向EphA2 (AstraZeneca；Seagen (Seattle Genetics) Inc.)；伏司妥珠單抗莫福汀，亦稱為SGN-75，其靶向CD70 (Seagen (Seattle Genetics) Inc.)；德甯土珠單抗莫福汀，亦稱為SGN-CD19A，其靶向CD19 (Seagen (Seattle Genetics) Inc.)及HTI-1066，亦稱為SHR-A1403，其靶向c-MET (Jiangsu HengRui Medicine Co., Ltd)。In some embodiments, the ADC comprises MMAF and is one of: CD70-ADC, which targets CD70 (Kochi University; Osaka University); IGN786, which targets SAIL (AstraZeneca; Igenica Biotherapeutics); PF-06263507, It targets 5T4 (Pfizer); GPC1-ADC, which targets GPC-1 (Kochi University); ADC-AVP10, which targets CD30 (Avipep); M290-MC-MMAF, which targets CD103 (The Second Affiliated Hospital of Harbin Medical University); BVX001, which targets CD33; CD7 (Bivictrix therapeutics); Tanabe P3D12-vc-MMAF, which targets c-MET (Tanabe Research Laboratories); LILRB4-targeting ADC, which targets LILRB4 (The University of Texas Health Science Center, Houston); TSD101, also known as ABL201, which targets BCMA (TSD Life Science; ABL Bio; Lego Chem Biosciences); Martin-dituxizumab mofotine, also known as ABT-414, which targets EGFR (Abbvie; Seagen (Seattle Genetics) Inc.); AGS16F, also known as AGS-16C3F; AGS-16M8F, which targets ENPP3 (Astellas Pharma Inc.; Seagen (Seattle Genetics) Inc.) ); AVG-A11 BCMA ADC, also known as AVG-A11-mcMMAF, which targets BCMA (Avantgen); belantimumab mofotin, also known as BLENREP; GSK2857916; J6M0-mcMMAF, which targets BCMA ( GlaxoSmithKline; Seagen (Seattle Genetics) Inc.); MP-HER3-ADC, also known as HER3-ADC, which targets HER-3 (MediaPharma); FS-1502, also known as LCB14-0110, which targets HER- 2 (Lego Chem Biosciences; Shanghai F osun Pharmaceutical Development Co, Ltd.); MEDI-547, also known as MI-CP177, which targets EphA2 (AstraZeneca; Seagen (Seattle Genetics) Inc.); vostuzumab mufotine, also known as SGN -75, which targets CD70 (Seagen (Seattle Genetics) Inc.); denizumab mofotenumab, also known as SGN-CD19A, which targets CD19 (Seagen (Seattle Genetics) Inc.) and HTI-1066, Also known as SHR-A1403, it targets c-MET (Jiangsu HengRui Medicine Co., Ltd).

在一些實施例中，ADC係選自表A、表B或表C中之ADC。在表A、表B及表C中，具有序列表中提供之序列的ADC用星號(*)標記。在一些實施例中，ADC不為恩諾單抗維多汀。在某些實施例中，ADC不為本妥昔單抗維多汀。在一些實施例中，ADC不為替索圖單抗維多汀。在一些實施例中，ADC不為拉地妥珠單抗維多汀。在一些實施例中，ADC不為SGN-CD228A。 表 A 專利 藥物名稱 目標 部分 連接子 有效負載 US 8,545,850 波妥珠單抗維多汀 CD79b 波妥珠單抗同種型：IgG1，來源：人類化 纈胺酸-瓜胺酸 MMAE    吉妥珠單抗奧唑米星 CD33 吉妥單抗同種型：IgG4，來源：人類化 AcBut醯基腙-雙硫鍵 卡奇黴素 US 9,273,141 貝蘭他單抗莫福汀 BCMA 貝蘭單抗(J6M0)同種型：IgG1，來源：人類化，格式：mAB mc MMAF US  9,808,537 曲妥珠單抗德魯特坎 HER-2 曲妥珠單抗同種型：IgG1，來源：人類化 GGFG (甘胺酸-甘胺酸-苯丙胺酸-甘胺酸) DXd/DX8951 (MAAA-1181a) US 8,637,642 (WO 2012/047724) 恩諾單抗維多汀* 連接素-4 恩諾單抗同種型：IgG1k，來源：人類 纈胺酸-瓜胺酸 MMAE US 8,153,768 英妥珠單抗奧佐米星 CD22 英妥珠單抗同種型：IgG4，來源：人類化 AcBut醯基腙-雙硫鍵 卡奇黴素 WO 2004/010957 本妥昔單抗維多汀(SGN-35)* CD30 本妥昔單抗同種型：IgG1，來源：嵌合 纈胺酸-瓜胺酸 MMAE US 7,517,964; US 8,877,901 薩西土珠單抗戈維特坎 TROP-2 薩西土珠單抗(hRS7)同種型：IgG1，來源：人類化，格式：mAB CL2A SN-38 US 8,337,856 曲妥珠單抗恩他新 HER-2 曲妥珠單抗同種型：IgG1，來源：人類化 SMCC DM1 表 B 藥物名稱 其他藥物名稱 目標 部分 連接子 有效負載 CX-2029 ABBV-2029 CD71 前抗體 纈胺酸-瓜胺酸 MMAE DP303c    HER-2    未知 MMAE HTI-1066 SHR-A1403 c-MET 抗C-MET同種型：IgG2，來源：人類化，格式：mAB 3-丙基硫-mc MMAF FOR46    CD46 23AG2同種型：IgG1，來源：人類，格式：mAB，其他：抗CD46人類化 纈胺酸-瓜胺酸 MMAF AGS62P1 ASP1235 FLT3 抗FLT3單株抗體同種型：IgG1，來源：人類 肟 AGD-0182 BT8009    連接素-4    纈胺酸-瓜胺酸 MMAE ARX788    HER-2    肟 Amberstatin269 XMT-1536    NaPi2b XMT-1535同種型：IgG1，來源：人類化，格式：mAB 弗萊西莫(Fleximer)聚合物 奧瑞他汀F-HPA ALT-P7 HM2-MMAE HER-2 曲妥珠單抗同種型：IgG1，來源：人類化，格式：mAB，其他：曲妥珠單抗之NexMab變異體 纈胺酸-瓜胺酸 MMAE FS-1502 LCB14-0110 HER-2 曲妥珠單抗 β-葡萄糖醛酸苷酶(BG)連接子 MMAF 考非妥珠單抗比利多汀 PF-06647020、PTK7-ADC、PF-7020、ABBV-647 PTK7 h6M24同種型：IgG1，來源：人類化，格式：mAB 纈胺酸-瓜胺酸 PF-06380101 TRS005    CD20 利妥昔單抗同種型：IgG1，來源：嵌合，格式：mAb 纈胺酸-瓜胺酸 MMAE STI-6129 CD38 ADC、LNDS1001、CD38-077 ADC CD38 STI-5171同種型：IgG1，來源：人類，格式：mAB 未知 斯他汀5.2 替索圖單抗維多汀* Humax-TF-ADC, tf-011-mmae CD142 HuMab (HuMax抗體)同種型：IgG1，來源：人類 纈胺酸-瓜胺酸 MMAE 瑟吐珠單抗維多汀 VLS-101, UC-961ADC3 ROR1 瑟吐珠單抗(UC-961)同種型：IgG1，來源：人類化，格式：mAB 可裂解蛋白酶 MMAE AbGn-107 Ab1-18Hr1 AG-7 AbGn-7同種型：IgG1，來源：人類化，格式：mAB 纈胺酸-瓜胺酸 MMAD ASN-004    5T4 未揭示格式：scFvFc抗體 弗萊西莫聚合物 尾海兔素 SGN-B6A*    整合素β-6 h2A2來源：人類化 纈胺酸-瓜胺酸 MMAE 迪西妥單抗維多汀* RC48 HER-2 赫妥珠單抗(Hertuzumab)同種型：IgG1，來源：人類化，格式：mAB 纈胺酸-瓜胺酸 MMAE 肽利索單抗維多汀 ABBV-399 c-MET ABT-700 纈胺酸-瓜胺酸 MMAE 依那波他單抗維多汀 HuMax-AXL-ADC, AXL-107-MMAE Axl HuMax抗體來源:人類，格式:全長 纈胺酸-瓜胺酸 MMAE OBI-999    Globo H OBI-888同種型：IgG1，來源：人類化，格式：mAB 二硫化物 MMAE XMT-1592    NaPi2b 未知 未知 Auristatin F-HPA SGN-CD228A    CD228 hL49 (抗CD228A單株抗體),來源：人類化，格式：mAB β-葡萄糖醛酸苷酶(BG)連接子 MMAE 拉地妥珠單抗維多汀* SGN-LIV1A LIV-1 拉地妥珠單抗(hLIV-22)同種型：IgG1，來源：人類化，格式：mAB 纈胺酸-瓜胺酸 MMAE BT5528    EphA2 未知 纈胺酸-瓜胺酸 MMAE PF-06804103 NG-HER2 ADC HER-2 未知同種型：IgG1，來源：人類 纈胺酸-瓜胺酸 PF-06380101 (Aur 101) W0101    IGF-1R hz208F2-4 (抗IGF1R抗體)，來源：人類化，格式：mAB mc 奧瑞他汀 BA3011 CAB-AXL-ADC Axl CAB-Axl其他：條件活性生物製劑(CAB)抗Axl抗體 未知 MMAE MRG002    HER-2 抗HER2同種型：IgG1，來源：人類化，格式：mAB 未知 MMAE ZW49    HER-2,HER-2 ZW25同種型：IgG1，格式：雙特異性，ZW25同種型：IgG1，格式：雙特異性 纈胺酸-瓜胺酸 奧瑞他汀 表 C 專利 ADC名稱 目標： US 9,168,314 (WO 2011/157741) 替索圖單抗維多汀(TV)* CD142 (組織因子) US 9,273,141 貝蘭他單抗莫福汀 BCMA US 8,637,642 (WO 2012/047724) 恩諾單抗維多汀(EV)* 連接素-4 US 11,028,181 (WO2017083582A1)；參見本文中的序列表 SGN-STNV STN    依那波他單抗維多汀 Axl US 2020/0246479 (WO 2020/163225；分別為SEQ ID NO: 7及8之VH/VL(CDR SEQ ID NO: 1-6))；參見本文中之序列表 SGN-CD228A* CD228 US 2021/0198367 (主張USSN 62/943,959及USSN 62/012,584之優先權)；參見本文中之序列表 SGN-B6A* 整合素β-6 WO 2012/078688 拉地妥珠單抗維多汀(LV)* LIV-1 WO 2004/010957 本妥昔單抗維多汀(SGN-35)* CD30 US 8,329,173 肽利索單抗維多汀 c-MET US 8,545,850 波妥珠單抗維多汀 CD79b US 7,662,387 伏司妥珠單抗莫福汀 CD70 D. 製備抗體 In some embodiments, the ADC is selected from ADCs in Table A, Table B, or Table C. In Table A, Table B, and Table C, ADCs with sequences provided in the Sequence Listing are marked with an asterisk (*). In some embodiments, the ADC is not ennozumab vedotin. In certain embodiments, the ADC is not bentuximab vedotin. In some embodiments, the ADC is not tesotumumab vedotin. In some embodiments, the ADC is not latstuzumab vedotin. In some embodiments, the ADC is not SGN-CD228A. Table A patent drug name Target part linker payload US 8,545,850 pertuzumab vedotin CD79b Perstuzumab Isotype: IgG1, Source: Humanized valine-citrulline MMAE gemtuzumab ozogamicin CD33 Gemtuzumab Isotype: IgG4, Source: Humanized AcBut acylhydrazone-disulfide bond calicheamicin US 9,273,141 belantimumab mofotine BCMA Belamumab (J6M0) Isotype: IgG1, Source: Humanized, Format: mAB mc MMAF US 9,808,537 Trastuzumab Drutcan HER-2 Trastuzumab Isotype: IgG1, Source: Humanized GGFG (glycine-glycine-phenylalanine-glycine) DXd/DX8951 (MAAA-1181a) US 8,637,642 (WO 2012/047724) ennozumab vedotin* connexin-4 Ennozumab Isotype: IgG1k, Source: Human valine-citrulline MMAE US 8,153,768 Intozumab ozogamicin CD22 Intozumab Isotype: IgG4, Source: Humanized AcBut acylhydrazone-disulfide bond calicheamicin WO 2004/010957 Bentuximab vedotin (SGN-35)* CD30 Bentuximab Isotype: IgG1, Source: Chimeric valine-citrulline MMAE US 7,517,964; US 8,877,901 Sassi Tuilizumab Govitkan TROP-2 Saxituzumab (hRS7) Isotype: IgG1, Source: Humanized, Format: mAB CL2A SN-38 US 8,337,856 Trastuzumab, Entacin HER-2 Trastuzumab Isotype: IgG1, Source: Humanized SMCC DM1 Form B drug name Other drug names Target part linker payload CX-2029 ABBV-2029 CD71 pre-antibody valine-citrulline MMAE DP303c HER-2 unknown MMAE HTI-1066 SHR-A1403 c-MET Anti-C-MET Isotype: IgG2, Source: Humanized, Format: mAB 3-propylthio-mc MMAF FOR46 CD46 23AG2 Isotype: IgG1, Source: Human, Format: mAB, Other: Anti-CD46 Humanized valine-citrulline MMAF AGS62P1 ASP1235 FLT3 Anti-FLT3 Monoclonal Antibody Isotype: IgG1, Source: Human oxime AGD-0182 BT8009 connexin-4 valine-citrulline MMAE ARX788 HER-2 oxime Amberstatin269 XMT-1536 NaPi2b XMT-1535 Isotype: IgG1, Source: Humanized, Format: mAB Fleximer polymers Auristatin F-HPA ALT-P7 HM2-MMAE HER-2 Trastuzumab Isotype: IgG1, Source: Humanized, Format: mAB, Other: NexMab Variant of Trastuzumab valine-citrulline MMAE FS-1502 LCB14-0110 HER-2 Trastuzumab β-glucuronidase (BG) linker MMAF Corfetuzumab Billidotine PF-06647020, PTK7-ADC, PF-7020, ABBV-647 PTK7 h6M24 Isotype: IgG1, Source: Humanized, Format: mAB valine-citrulline PF-06380101 TRS005 CD20 Rituximab Isotype: IgG1, Source: Chimeric, Format: mAb valine-citrulline MMAE STI-6129 CD38 ADC, LNDS1001, CD38-077 ADC CD38 STI-5171 Isotype: IgG1, Source: Human, Format: mAB unknown statin 5.2 tesotuzumab vedotin* Humax-TF-ADC, tf-011-mmae CD142 HuMab (HuMax Antibody) Isotype: IgG1, Source: Human valine-citrulline MMAE cetuzumab vedotin VLS-101, UC-961ADC3 ROR1 Cetumuzumab (UC-961) Isotype: IgG1, Source: Humanized, Format: mAB cleavable protease MMAE AbGn-107 Ab1-18Hr1 AG-7 AbGn-7 Isotype: IgG1, Source: Humanized, Format: mAB valine-citrulline MMAD ASN-004 5T4 Undisclosed format: scFvFc antibody Fresimo Polymers Aplysia SGN-B6A* integrin beta-6 h2A2 source: humanized valine-citrulline MMAE dicituzumab vedotin* RC48 HER-2 Hertuzumab Isotype: IgG1, Source: Humanized, Format: mAB valine-citrulline MMAE Peptide Lisomab Vidotin ABBV-399 c-MET ABT-700 valine-citrulline MMAE enabolatumab vedotin HuMax-AXL-ADC, AXL-107-MMAE Axl HuMax Antibody Source: Human, Format: Full Length valine-citrulline MMAE OBI-999 Globo H OBI-888 Isotype: IgG1, Source: Humanized, Format: mAB Disulfide MMAE XMT-1592 NaPi2b unknown unknown Auristatin F-HPA SGN-CD228A CD228 hL49 (anti-CD228A monoclonal antibody), source: humanized, format: mAB β-glucuronidase (BG) linker MMAE Ladetuzumab vedotin* SGN-LIV1A LIV-1 Ladetuzumab (hLIV-22) Isotype: IgG1, Source: Humanized, Format: mAB valine-citrulline MMAE BT5528 EphA2 unknown valine-citrulline MMAE PF-06804103 NG-HER2 ADCs HER-2 Unknown isotype: IgG1, source: human valine-citrulline PF-06380101 (Aur 101) W0101 IGF-1R hz208F2-4 (anti-IGF1R antibody), source: humanized, format: mAB mc Auristatin BA3011 CAB-AXL-ADC Axl CAB-Axl Other: Conditionally Active Biologics (CAB) Anti-Axl Antibody unknown MMAE MRG002 HER-2 Anti-HER2 Isotype: IgG1, Source: Humanized, Format: mAB unknown MMAE ZW49 HER-2,HER-2 ZW25 Isotype: IgG1, Format: Bispecific, ZW25 Isotype: IgG1, Format: Bispecific valine-citrulline Auristatin Form C patent ADC name Target: US 9,168,314 (WO 2011/157741) tesotuzumab vedotin (TV)* CD142 (tissue factor) US 9,273,141 belantimumab mofotine BCMA US 8,637,642 (WO 2012/047724) ennozumab vedotin (EV)* connexin-4 US 11,028,181 (WO2017083582A1); see Sequence Listing herein SGN-STNV STN enabolatumab vedotin Axl US 2020/0246479 (WO 2020/163225; VH/VL of SEQ ID NOs: 7 and 8, respectively (CDR SEQ ID NOs: 1-6)); see Sequence Listing herein SGN-CD228A* CD228 US 2021/0198367 (claims priority to USSN 62/943,959 and USSN 62/012,584); see Sequence Listing herein SGN-B6A* integrin beta-6 WO 2012/078688 Ladetuzumab vedotin (LV)* LIV-1 WO 2004/010957 Bentuximab vedotin (SGN-35)* CD30 US 8,329,173 Peptide Lisomab Vidotin c-MET US 8,545,850 pertuzumab vedotin CD79b US 7,662,387 vostuzumab mofotine CD70 D. Preparation of Antibodies

為製備抗體，可使用此項技術中已知之許多技術。 參見例如Kohler & Milstein， Nature256:495-497 (1975)；Kozbor 等人， Immunology Today4: 72 (1983)；Cole 等人， Monoclonal Antibodies and Cancer Therapy中之第77-96頁，Alan R. Liss, Inc. (1985)；Coligan， Current Protocols in Immunology(1991)；Harlow & Lane， Antibodies, A Laboratory Manual(1988)；以及Goding， Monoclonal Antibodies: Principles and Practice(第2版，1986)。 To prepare antibodies, many techniques known in the art can be used. See, eg , Kohler & Milstein, Nature 256:495-497 (1975); Kozbor et al., Immunology Today 4:72 (1983); Cole et al., pp. 77-96 in Monoclonal Antibodies and Cancer Therapy , Alan R. Liss , Inc. (1985); Coligan, Current Protocols in Immunology (1991); Harlow & Lane, Antibodies, A Laboratory Manual (1988); and Goding, Monoclonal Antibodies: Principles and Practice (2nd ed., 1986).

編碼所關注之抗體之重鏈及輕鏈的基因可自細胞選殖，例如編碼單株抗體之基因可自表現該抗體之融合瘤選殖且用於產生重組單株抗體。編碼單株抗體之重鏈及輕鏈的基因庫亦可由融合瘤或漿細胞製得。另外，噬菌體或酵母菌呈現技術可用於鑑別特異性結合於選定抗原之抗體及異質Fab片段( 參見例如McCafferty 等人 ， Nature348:552-554 (1990)；Marks 等人 ， Biotechnology10:779-783 (1992)；Lou 等人 ，(2010) PEDS23:311；以及Chao 等人， Nature Protocols,1:755-768 (2006))。替代地，抗體及抗體序列可使用基於酵母菌之抗體呈遞系統分離及/或鑑別，該抗體呈遞系統諸如揭示於例如Xu 等人 ， Protein Eng Des Sel,2013, 26:663-670；WO 2009/036379；WO 2010/105256；以及WO 2012/009568中之抗體呈遞系統。重鏈及輕鏈基因產物之隨機組合產生具有不同抗原特異性之大抗體池(參見例如Kuby, Immunology(第3版，1997))。用於產生單鏈抗體或重組抗體之技術(美國專利4,946,778、美國專利第4,816,567號)亦可適用於產生抗體。抗體亦可經製成為雙特異性，亦即能夠識別兩種不同抗原( 參見例如WO 93/08829，Traunecker 等人 ， EMBO J.10:3655-3659 (1991)；以及Suresh 等人 ， Methods in Enzymology121:210 (1986))。抗體亦可為雜結合物，例如兩種共價接合的抗體，或共價結合於免疫毒素之抗體( 參見例如美國專利第4,676,980號、WO 91/00360；以及WO 92/200373)。 Genes encoding the heavy and light chains of an antibody of interest can be cloned from cells, eg, a gene encoding a monoclonal antibody can be cloned from a fusion tumor expressing the antibody and used to produce recombinant monoclonal antibody. Gene repertoires encoding the heavy and light chains of monoclonal antibodies can also be made from fusionomas or plasma cells. Additionally, phage or yeast display techniques can be used to identify antibodies and heterogeneous Fab fragments that specifically bind to selected antigens ( see, eg , McCafferty et al ., Nature 348:552-554 (1990); Marks et al ., Biotechnology 10:779-783 (1992); Lou et al , (2010) PEDS 23:311; and Chao et al, Nature Protocols, 1:755-768 (2006)). Alternatively, antibodies and antibody sequences can be isolated and/or identified using a yeast-based antibody presentation system such as disclosed for example in Xu et al ., Protein Eng Des Sel, 2013, 26:663-670; WO 2009/ 036379; WO 2010/105256; and antibody presentation systems in WO 2012/009568. Random combinations of heavy and light chain gene products generate large pools of antibodies with different antigen specificities (see, eg, Kuby, Immunology (3rd ed., 1997)). Techniques used to generate single chain antibodies or recombinant antibodies (US Pat. No. 4,946,778, US Pat. No. 4,816,567) can also be adapted to generate antibodies. Antibodies can also be made bispecific, that is, capable of recognizing two different antigens ( see, eg , WO 93/08829, Traunecker et al. , EMBO J. 10:3655-3659 (1991); and Suresh et al ., Methods in Enzymology 121:210 (1986)). Antibodies can also be heteroconjugates, such as two covalently joined antibodies, or antibodies covalently bound to an immunotoxin ( see, eg , US Pat. No. 4,676,980, WO 91/00360; and WO 92/200373).

可使用任何數目的表現系統，包括原核表現系統及真核表現系統來產生抗體。在一些實施例中，表現系統為哺乳動物細胞，諸如融合瘤，或CHO細胞。許多此類系統可自商業供應商廣泛購得。在其中抗體包含重鏈及輕鏈兩者之實施例中，例如在雙順反子表現單元中或在不同啟動子的控制下，重鏈及重鏈以及輕鏈可使用單一載體表現。在其他實施例中，重鏈及輕鏈區可使用個別載體表現。如本文所描述之重鏈及輕鏈可視情況在N末端處包含甲硫胺酸。 Antibodies can be produced using any number of expression systems, including prokaryotic and eukaryotic expression systems. In some embodiments, the expression system is a mammalian cell, such as a fusionoma, or a CHO cell. Many such systems are widely available from commercial suppliers. In embodiments in which the antibody comprises both heavy and light chains, eg, in a bicistronic expression unit or under the control of different promoters, the heavy chain and the heavy and light chains can be expressed using a single vector. In other embodiments, the heavy and light chain regions can be expressed using separate vectors. Heavy and light chains as described herein may optionally contain methionine at the N-terminus.

在一些實施例中，產生抗體片段(諸如Fab、Fab'、F(ab') 2、scFv或雙功能抗體)。已研發出用於產生抗體片段之各種技術。傳統上，此等片段經由完整抗體之蛋白分解消化而衍生( 參見例如Morimoto 等人 ， J. Biochem. Biophys. Meth., 24:107-117 (1992)；以及Brennan 等人 ， Science,229:81 (1985))。然而，此等片段現可使用重組宿主細胞直接產生。舉例而言，抗體片段可自抗體噬菌體庫分離。替代地，Fab'-SH片段可自大腸桿菌(E. coli)細胞直接回收且經化學偶合以形成F(ab') 2片段( 參見例如Carter 等人 ， BioTechnology, 10:163-167 (1992))。根據另一方法，F(ab') 2片段可自重組宿主細胞培養物直接分離。用於產生抗體片段之其他技術將為熟習此項技術者顯而易見的。在其他實施例中，所選抗體為單鏈Fv片段(scFv)。 參見例如PCT公開案第WO 93/16185號；以及美國專利第5,571,894號及第5,587,458號。該抗體片段亦可為如例如美國專利第5,641,870號中所描述之直鏈抗體。 In some embodiments, antibody fragments (such as Fab, Fab', F(ab') 2 , scFv, or diabodies) are produced. Various techniques have been developed for producing antibody fragments. Traditionally, such fragments are derived via proteolytic digestion of intact antibodies ( see, eg , Morimoto et al ., J. Biochem. Biophys. Meth. , 24:107-117 (1992); and Brennan et al ., Science, 229:81 (1985)). However, these fragments can now be produced directly using recombinant host cells. For example, antibody fragments can be isolated from antibody phage libraries. Alternatively, Fab'-SH fragments can be recovered directly from E. coli cells and chemically coupled to form F(ab') 2 fragments ( see, eg , Carter et al ., BioTechnology , 10:163-167 (1992) ). According to another method, F(ab') 2 fragments can be isolated directly from recombinant host cell culture. Other techniques for producing antibody fragments will be apparent to those skilled in the art. In other embodiments, the antibody of choice is a single chain Fv fragment (scFv). See, eg , PCT Publication No. WO 93/16185; and US Patent Nos. 5,571,894 and 5,587,458. The antibody fragment can also be a linear antibody as described, for example, in US Pat. No. 5,641,870.

在一些實施例中，抗體或抗體片段可與另一分子，例如聚乙二醇(PEG化)或血清白蛋白結合，以提供延長的活體內半衰期。抗體片段之PEG化實例提供於Knigh 等人 ， Platelets15:409, 2004 (針對阿昔單抗(abciximab))；Pedley 等人 ， Br. J. Cancer70:1126, 1994 (針對抗CEA抗體)；Chapman 等人， Nature Biotech.17:780, 1999；以及Humphreys 等人， Protein Eng. Des.20: 227, 2007)。 In some embodiments, the antibody or antibody fragment can be conjugated to another molecule, such as polyethylene glycol (PEGylated) or serum albumin, to provide extended in vivo half-life. Examples of PEGylation of antibody fragments are provided in Knigh et al ., Platelets 15:409, 2004 (for abciximab); Pedley et al ., Br. J. Cancer 70:1126, 1994 (for anti-CEA antibodies); Chapman et al, Nature Biotech. 17:780, 1999; and Humphreys et al, Protein Eng. Des. 20: 227, 2007).

在一些實施例中，提供多特異性抗體，例如雙特異性抗體。多特異性抗體為對至少兩種不同抗原或對同一抗原之至少兩個不同抗原決定基具有結合特異性之抗體。用於製得多特異性抗體之方法包括但不限於在宿主細胞中重組共表現兩對重鏈及輕鏈(參見例如Zuo等人， Protein Eng Des Sel,2000, 13:361-367)；「杵-臼」工程化(參見例如Ridgway等人， Protein Eng Des Sel,1996, 9:617-721)；「雙功能抗體」技術(參見例如Hollinger等人， PNAS (USA), 1993, 90:6444-6448)；以及分子內三聚(參見例如Alvarez-Cienfuegos等人， Scientific Reports, 2016, 數位物件識別碼：/10.1038/srep28643)；亦參見Spiess等人， Molecular Immunology, 2015, 67(2), 部分A：95-106。 恆定區之選擇 In some embodiments, multispecific antibodies, such as bispecific antibodies, are provided. Multispecific antibodies are antibodies that have binding specificities for at least two different antigens or for at least two different epitopes of the same antigen. Methods for making multispecific antibodies include, but are not limited to, recombinantly co-expressing two pairs of heavy and light chains in a host cell (see, e.g., Zuo et al., Protein Eng Des Sel, 2000, 13:361-367);"Knob-hole" engineering (see eg Ridgway et al, Protein Eng Des Sel, 1996, 9:617-721); "Diabody" technology (see eg Hollinger et al, PNAS (USA) , 1993, 90:6444 -6448); and intramolecular trimerization (see, e.g., Alvarez-Cienfuegos et al., Scientific Reports , 2016, DID: /10.1038/srep28643); see also Spiess et al., Molecular Immunology , 2015, 67(2), Part A: 95-106. Choice of constant region

本文所描述之抗體之重鏈及輕鏈可變區可鍵聯至人類恆定區的至少一部分。恆定區之選擇可部分地視是否需要抗體依賴性細胞介導之細胞毒性、抗體依賴性細胞吞噬作用及/或補體依賴性細胞毒性而定。舉例而言，人類同位素IgG1及IgG3具有較強補體依賴性細胞毒性，人類同種型IgG2弱補體依賴性細胞毒性且人類IgG4缺乏補體依賴性細胞毒性。另外，人類IgG1及IgG3誘導比人類IgG2及IgG4更強的細胞介導之效應功能。輕鏈恆定區可為λ或κ。抗體可表現為含有兩條輕鏈及兩條重鏈之四聚體，表現為單獨的重鏈、輕鏈，表現為Fab、Fab'、F(ab') 2及Fv，或表現為單鏈抗體，其中重鏈及輕鏈可變域經由間隔子連接。 The heavy and light chain variable regions of the antibodies described herein can be linked to at least a portion of a human constant region. The choice of constant region may depend in part on whether antibody-dependent cell-mediated cytotoxicity, antibody-dependent cellular phagocytosis, and/or complement-dependent cytotoxicity are desired. For example, the human isotopes IgGl and IgG3 have strong complement-dependent cytotoxicity, the human isotype IgG2 has weak complement-dependent cytotoxicity, and human IgG4 lacks complement-dependent cytotoxicity. In addition, human IgG1 and IgG3 induce stronger cell-mediated effector functions than human IgG2 and IgG4. The light chain constant region can be lambda or kappa. Antibodies may appear as tetramers containing two light chains and two heavy chains, as separate heavy chains, light chains, as Fab, Fab', F(ab') 2 and Fv, or as single chains An antibody in which the heavy and light chain variable domains are linked via a spacer.

人類恆定區展示不同個體之間的同種異型變異及異種異型變異，亦即，恆定區可在不同個體中的一或多個多態位置處不同。異種異型不同於同種異型之處在於血清識別同種異型與一或多個其他同型之非多態區結合。Human constant regions exhibit both allovariation and allovariation between individuals, ie, the constant regions may differ at one or more polymorphic positions in different individuals. Allotypes differ from allotypes in that serum recognizes allotypes in association with non-polymorphic regions of one or more other isotypes.

輕鏈及/或重鏈之胺基或羧基端之一個或若干個胺基酸，諸如重鏈之C端離胺酸，可缺失或在一定比例或全部的分子中衍生。取代可在恆定區中進行以降低或增大效應功能，諸如補體介導之細胞毒性或ADCC (參見例如Winter等人，美國專利第5,624,821號；Tso等人，美國專利第5,834,597號；及Lazar等人, Proc. Natl. Acad. Sci. USA 103:4005, 2006)，或以延長在人類中之半衰期(參見例如Hinton等人, J. Biol. Chem. 279:6213, 2004)。One or several amino acids at the amino or carboxyl terminus of the light chain and/or the heavy chain, such as the C-terminal lysine of the heavy chain, may be deleted or derivatized in a proportion or all of the molecule. Substitutions can be made in the constant region to reduce or increase effector functions, such as complement-mediated cytotoxicity or ADCC (see, eg, Winter et al., US Patent No. 5,624,821; Tso et al., US Patent No. 5,834,597; and Lazar et al. Human, Proc. Natl. Acad. Sci. USA 103:4005, 2006), or to prolong half-life in humans (see, eg, Hinton et al., J. Biol. Chem. 279:6213, 2004).

為構築所需抗體-藥物結合物，在一些實施例中，在胺基酸位置234、235、237、239、267、298、299、326、330或332處引入包括天然胺基酸取代成半胱胺酸殘基之胺基酸取代的例示性取代，較佳地人類IgG1同型中之S239C突變(編號係根據EU索引進行(Kabat， Sequences of Proteins of Immunological Interest(National Institutes of Health, Bethesda, MD, 1987及1991)；參見US 20100158909，其以引用的方式併入本文中)。額外半胱胺酸殘基之存在可允許鏈間二硫鍵形成。此類鏈間二硫鍵形成可導致位阻，進而減小Fc區-FcγR結合相互作用之親和力。引入IgG恆定區之Fc區中或附近之一或多個半胱胺酸殘基亦可充當用於結合治療劑之位點(亦即，使用硫醇特異性試劑，諸如藥物之順丁烯二醯亞胺衍生物來偶合細胞毒性藥物)。治療劑之存在導致位阻，進而進一步減小Fc區-FcγR結合相互作用之親和力。位置234、235、236及/或237中之任一者處的其他取代減小對於Fcγ受體(特定言之，FcγRI受體)之親和力( 參見例如US 6,624,821，US 5,624,821)。 To construct the desired antibody-drug conjugates, in some embodiments, at amino acid positions 234, 235, 237, 239, 267, 298, 299, 326, 330, or 332, the introduction includes substitution of natural amino acids into half Exemplary substitutions of amino acid substitutions of cystine residues, preferably the S239C mutation in the human IgG1 isotype (numbering is according to the EU index (Kabat, Sequences of Proteins of Immunological Interest (National Institutes of Health, Bethesda, MD) , 1987 and 1991); see US 20100158909, which is incorporated herein by reference). The presence of additional cysteine residues may allow for interchain disulfide bond formation. Such interchain disulfide bond formation may lead to This reduces the affinity of the Fc region-FcyR binding interaction. One or more cysteine residues introduced into or near the Fc region of the IgG constant region can also serve as a site for binding of therapeutic agents (i.e. , using thiol-specific reagents such as maleimide derivatives of the drug to couple cytotoxic drugs). The presence of the therapeutic agent results in steric hindrance, which in turn further reduces the affinity of the Fc region-FcγR binding interaction. Position Other substitutions at any of 234, 235, 236 and/or 237 reduce affinity for Fcγ receptors (specifically, FcγRI receptors) ( see eg US 6,624,821, US 5,624,821).

抗體之活體內半衰期亦可對其效應功能產生影響。可增加或降低抗體之半衰期以調節其治療活性。FcRn為結構上類似於與β2-微球蛋白非共價締合之I類MHC抗原的受體。FcRn調節IgG之代謝及其在整個組織中之胞吞轉運(Ghetie及Ward, 2000, Annu. Rev. Immunol.18:739-766；Ghetie及Ward, 2002, Immunol. Res.25:97-113)。IgG-FcRn相互作用發生在pH 6.0 (細胞內小泡之pH)而非pH 7.4 (血液之pH)下；此相互作用能夠使得IgG再循環回至循環(Ghetie及Ward, 2000, Ann. Rev. Immunol.18:739-766；Ghetie及Ward, 2002, Immunol. Res.25:97-113)。已定位人類IgG1上參與FcRn結合之區(Shields 等人 ,2001, J. Biol. Chem.276:6591-604)。人類IgG1之位置Pro238、Thr256、Thr307、Gln311、Asp312、Glu380、Glu382或Asn434處之丙胺酸取代增強FcRn結合(Shields 等人 ,2001, J. Biol. Chem.276:6591-604)。具有此等取代之IgG1分子具有更長血清半衰期。因此，與未經修飾之IgG1相比，此等經修飾IgG1分子可能夠在更長時間段內進行其效應功能，且因此發揮其治療功效。用於增加與FcRn之結合的其他例示性取代包括250位置處之Gln及/或位置428處之Leu。EU編號用於恆定區中之所有位置。 The in vivo half-life of an antibody can also have an impact on its effector function. The half-life of an antibody can be increased or decreased to modulate its therapeutic activity. FcRn is a receptor that is structurally similar to MHC class I antigens non-covalently associated with β2-microglobulin. FcRn regulates the metabolism of IgG and its transcytosis throughout tissues (Ghetie and Ward, 2000, Annu. Rev. Immunol. 18:739-766; Ghetie and Ward, 2002, Immunol. Res. 25:97-113) . The IgG-FcRn interaction occurs at pH 6.0 (pH of intracellular vesicles) rather than pH 7.4 (pH of blood); this interaction enables recycling of IgG back to the circulation (Ghetie and Ward, 2000, Ann. Rev. Immunol. 18:739-766; Ghetie and Ward, 2002, Immunol. Res. 25:97-113). The region on human IgG1 involved in FcRn binding has been mapped (Shields et al ., 2001, J. Biol. Chem. 276:6591-604). Alanine substitutions at positions Pro238, Thr256, Thr307, Gln311, Asp312, Glu380, Glu382 or Asn434 of human IgG1 enhanced FcRn binding (Shields et al ., 2001, J. Biol. Chem. 276:6591-604). IgGl molecules with these substitutions have longer serum half-lives. Thus, these modified IgGl molecules may be able to perform their effector functions over a longer period of time than unmodified IgGl, and thus exert their therapeutic efficacy. Other exemplary substitutions for increasing binding to FcRn include Gln at position 250 and/or Leu at position 428. EU numbers are used for all positions in the constant region.

抗體之補體結合活性(C1q結合及CDC活性兩者)可藉由Lys326及Glu333處之取代而提高(Idusogie 等人，2001 , J. Immunol.166:2571-2575)。人類IgG2主鏈上之相同取代可將與C1q不充分結合且嚴重缺乏補體活化活性之抗體同型轉化成可結合C1q且調節CDC之抗體同型(Idusogie 等人，2001, J. Immunol.166:2571-75)。若干其他方法亦已應用於提高抗體之補體結合活性。舉例而言，將IgM之18-胺基酸羧基端尾片移植至IgG之羧基端極大地增強其CDC活性。即使在通常不具有可偵測CDC活性之IgG4情況下亦觀測到此增強(Smith 等人, 1995, J. Immunol.154:2226-36)。此外，用Cys取代靠近IgG1重鏈之羧基端安置的Ser444誘導IgG1之尾部與尾部二聚合的CDC活性相比於單體IgG1增加200倍(Shopes 等人，1992, J. Immunol.148:2918-22)。另外，具有針對C1q之特異性的雙特異性雙功能抗體構築體亦賦予CDC活性(Kontermann 等人，1997, Nat. Biotech.15:629-31)。 The complement-fixing activity of the antibody (both C1q binding and CDC activity) can be enhanced by substitutions at Lys326 and Glu333 (Idusogie et al ., 2001, J. Immunol. 166:2571-2575). Identical substitutions on the human IgG2 backbone can convert an antibody isotype that binds insufficiently to C1q and severely lacks complement-activating activity to an antibody isotype that binds C1q and modulates CDC (Idusogie et al ., 2001, J. Immunol. 166:2571- 75). Several other methods have also been applied to increase the complement-fixing activity of antibodies. For example, grafting the 18-amino acid carboxy-terminal tail of IgM to the carboxy-terminus of IgG greatly enhanced its CDC activity. This enhancement was observed even with IgG4, which normally does not have detectable CDC activity (Smith et al ., 1995, J. Immunol. 154:2226-36). Furthermore, substitution of Cys for Ser444 positioned near the carboxy terminus of the IgG1 heavy chain induces a 200-fold increase in the CDC activity of tail-to-tail dimerization of IgG1 compared to monomeric IgG1 (Shopes et al ., 1992, J. Immunol. 148:2918- twenty two). In addition, bispecific diabody constructs with specificity for C1q also confer CDC activity (Kontermann et al ., 1997, Nat. Biotech. 15:629-31).

補體活性可藉由使重鏈之胺基酸殘基318、320及322中之至少一者突變成具有不同側鏈之殘基(諸如Ala)而降低。其他經烷基取代之非離子型殘基，諸如Gly、Ile、Leu或Val，或替代三個殘基中之任一者的諸如Phe、Tyr、Trp及Pro之此類芳族非極性殘基亦降低或消除C1q結合。在殘基320及322而非318處可使用Ser、Thr、Cys及Met以降低或消除C1q結合活性。藉由極性殘基替代318 (Glu)殘基可調節而非消除C1q結合活性。用Ala替代殘基297 (Asn)使得移除裂解活性但僅稍微降低(弱約三倍)針對C1q之親和力。此改變破壞糖基化位點及補體活化所需之碳水化合物之存在。此位點處之任何其他取代亦破壞糖基化位點。以下突變及其任何組合亦降低C1q結合：D270A、K322A、P329A及P31IS (參見WO 06/036291)。Complement activity can be reduced by mutating at least one of amino acid residues 318, 320, and 322 of the heavy chain to residues with different side chains, such as Ala. Other alkyl-substituted non-ionic residues such as Gly, Ile, Leu or Val, or aromatic non-polar residues such as Phe, Tyr, Trp and Pro that replace any of the three residues Also reduces or eliminates C1q binding. Ser, Thr, Cys and Met can be used at residues 320 and 322 instead of 318 to reduce or eliminate CIq binding activity. Replacing the 318 (Glu) residue by a polar residue modulates, but not eliminates, C1q binding activity. Replacing residue 297 (Asn) with Ala removed cleavage activity but only slightly (about three-fold weaker) the affinity for C1q. This change disrupts glycosylation sites and the presence of carbohydrates required for complement activation. Any other substitution at this site also destroys the glycosylation site. The following mutations and any combination thereof also reduced CIq binding: D270A, K322A, P329A and P31IS (see WO 06/036291).

對人類恆定區之提及包括具有任何天然同種異型或天然同種異型中佔據多態性位置之殘基之任何排列的恆定區。此外，相對於天然人類恆定區可存在至多1、2、5或10個突變，諸如上文所指示之彼等突變以降低Fcγ受體結合或增加與FcRN之結合。 核酸、載體及宿主細胞 Reference to a human constant region includes constant regions having any native allotype or any arrangement of residues occupying a polymorphic position in a native allotype. Furthermore, up to 1, 2, 5 or 10 mutations, such as those indicated above, may be present relative to the native human constant region to decrease Fcγ receptor binding or increase binding to FcRN. Nucleic acids, vectors and host cells

在一些實施例中，本文所描述之抗體係使用重組方法製備。因此，在一些態樣中，本發明提供經分離核酸，其包含編碼本文所描述之任何抗體(例如本文所描述之CDR中之任一者或多者)之核酸序列；包含此類核酸之載體；以及宿主細胞，其中引入用於複製編碼抗體之核酸及/或表現抗體之核酸。在一些實施例中，宿主細胞為真核，例如中國倉鼠卵巢(CHO)細胞或人類細胞。In some embodiments, the antibody systems described herein are prepared using recombinant methods. Accordingly, in some aspects, the present invention provides isolated nucleic acids comprising nucleic acid sequences encoding any of the antibodies described herein (eg, any one or more of the CDRs described herein); vectors comprising such nucleic acids and host cells into which are introduced for replication of nucleic acid encoding the antibody and/or nucleic acid expressing the antibody. In some embodiments, the host cells are eukaryotic, such as Chinese Hamster Ovary (CHO) cells or human cells.

在一些實施例中，聚核苷酸(例如經分離聚核苷酸)包含編碼本文所描述之抗體之核苷酸序列。在一些實施例中，聚核苷酸包含編碼本文中所揭示之一或多種胺基酸序列(例如CDR、重鏈、輕鏈及/或構架區)之核苷酸序列。在一些實施例中，聚核苷酸包含編碼與本文中所揭示之序列(例如CDR、重鏈、輕鏈或構架區序列)具有至少85%序列一致性(例如至少85%、至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%序列一致性)之胺基酸序列的核苷酸序列。In some embodiments, the polynucleotides (eg, isolated polynucleotides) comprise nucleotide sequences encoding the antibodies described herein. In some embodiments, a polynucleotide comprises a nucleotide sequence encoding one or more of the amino acid sequences disclosed herein (eg, CDRs, heavy chains, light chains, and/or framework regions). In some embodiments, the polynucleotide comprises an encoding sequence that has at least 85% sequence identity (eg, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity) of the nucleotide sequence of the amino acid sequence.

在另一態樣中，提供製得本文所描述之抗體之方法。在一些實施例中，該方法包括在適用於表現抗體之條件下培養如本文所描述之宿主細胞(例如如本文所描述之表現聚核苷酸或載體的宿主細胞)。在一些實施例中，隨後自宿主細胞(或宿主細胞培養基)回收抗體。In another aspect, methods of making the antibodies described herein are provided. In some embodiments, the method comprises culturing a host cell as described herein (eg, a host cell expressing a polynucleotide or a vector as described herein) under conditions suitable for expressing the antibody. In some embodiments, the antibody is subsequently recovered from the host cell (or host cell culture medium).

含有編碼本發明之抗體的聚核苷酸或其片段的適合之載體包括選殖載體及表現載體。儘管所選擇的選殖載體可根據意欲使用之宿主細胞變化，但適用選殖載體一般能夠自我複製，可具有特定限制性核酸內切酶之單一目標及/或可攜帶可用於選擇含有載體之純系的標記物之基因。實例包括質體及細菌病毒，例如pUC18、pUC19、Bluescript (例如pBS SK+)及其衍生物、mpl8、mpl9、pBR322、pMB9、ColE1、pCR1、RP4、噬菌體DNA及穿梭載體(shuttle vector)，諸如pSA3及pAT28。選殖載體購自商業供應商，諸如BioRad、Stratagene及Invitrogen。Suitable vectors containing polynucleotides or fragments thereof encoding the antibodies of the invention include cloning vectors and expression vectors. Although the selection vector for selection may vary depending on the host cell intended to be used, suitable selection vectors are generally capable of self-replication, may have a single target for a particular restriction endonuclease, and/or may carry a clone that may be used to select for a vector-containing vector marker genes. Examples include plastids and bacterial viruses such as pUC18, pUC19, Bluescript (eg pBS SK+) and derivatives thereof, mpl8, mpl9, pBR322, pMB9, ColE1, pCR1, RP4, phage DNA and shuttle vectors such as pSA3 and pAT28. Colony vectors were purchased from commercial suppliers such as BioRad, Stratagene and Invitrogen.

表現載體一般為含有本發明之核酸的可複製聚核苷酸構築體。表現載體可在宿主細胞中以游離基因體或染色體DNA之整體部分形式複製。適合之表現載體包括但不限於質體、病毒載體，包括腺病毒、腺相關病毒、反轉錄病毒及任何其他載體。 重組抗體之表現 Expression vectors are typically replicable polynucleotide constructs containing the nucleic acids of the invention. Expression vectors can be replicated in host cells as episomal bodies or as integral parts of chromosomal DNA. Suitable expression vectors include, but are not limited to, plastids, viral vectors, including adenoviruses, adeno-associated viruses, retroviruses, and any other vector. Performance of recombinant antibodies

抗體通常藉由重組表現產生。重組聚核苷酸構築體通常包括可操作地連接於抗體鏈之編碼序列的表現控制序列，包括天然相關或異源啟動子區。較佳地，表現控制序列為載體中能夠轉型或轉染真核宿主細胞之真核啟動子系統。一旦載體已併入至適當宿主中，宿主就維持在適合高水準表現核苷酸序列且收集及純化交叉反應抗體的條件下。Antibodies are usually produced by recombinant expression. Recombinant polynucleotide constructs typically include expression control sequences operably linked to the coding sequence of the antibody chain, including naturally associated or heterologous promoter regions. Preferably, the expression control sequences are eukaryotic promoter systems in the vector capable of transforming or transfecting eukaryotic host cells. Once the vector has been incorporated into an appropriate host, the host is maintained under conditions suitable for high-level expression of the nucleotide sequence and collection and purification of cross-reacting antibodies.

哺乳動物細胞為用於表現編碼免疫球蛋白或其片段之核苷酸區段的較佳宿主。參見Winnacker , From Genes to Clones, (VCH Publishers, NY, 1987)。能夠分泌完整異源蛋白質之多個適合之宿主細胞株已在此項技術中開發，且包括CHO細胞株(例如DG44)、各種COS細胞株、HeLa細胞、HEK293細胞、L細胞及非抗體產生骨髓瘤，包括Sp2/0及NS0。較佳地，細胞為非人類的。用於此等細胞之表現載體可包括表現控制序列，諸如複製起點、啟動子、強化子(Queen等人, Immunol. Rev.89:49 (1986))，及必需的處理資訊位點，諸如核糖體結合位點、RNA剪接位點、聚腺苷酸化位點及轉錄終止子序列。較佳表現控制序列為來源於內源基因、巨細胞病毒、SV40、腺病毒、牛乳突狀瘤病毒及類似物之啟動子。參見Co等人, J. Immunol.148:1149 (1992)。 Mammalian cells are preferred hosts for expressing nucleotide segments encoding immunoglobulins or fragments thereof. See Winnacker , From Genes to Clones , (VCH Publishers, NY, 1987). A number of suitable host cell lines capable of secreting intact heterologous proteins have been developed in the art and include CHO cell lines (eg DG44), various COS cell lines, HeLa cells, HEK293 cells, L cells and non-antibody producing bone marrow Tumors, including Sp2/0 and NSO. Preferably, the cells are non-human. Expression vectors for use in such cells may include expression control sequences such as origins of replication, promoters, enhancers (Queen et al., Immunol. Rev. 89:49 (1986)), and sites for necessary processing information such as ribose body binding site, RNA splicing site, polyadenylation site and transcription terminator sequence. Preferred performance control sequences are promoters derived from endogenous genes, cytomegalovirus, SV40, adenovirus, bovine papilloma virus, and the like. See Co et al, J. Immunol. 148:1149 (1992).

一旦表現，抗體就可根據此項技術之標準程序純化，包括HPLC純化、管柱層析、凝膠電泳及其類似物(通常參見Scopes, Protein Purification (Springer-Verlag, NY, 1982))。 抗體表徵 Once expressed, antibodies can be purified according to standard procedures in the art, including HPLC purification, column chromatography, gel electrophoresis, and the like (see generally Scopes, Protein Purification (Springer-Verlag, NY, 1982)). Antibody characterization

用於分析結合親和力、結合動力學及交叉反應性之方法為此項技術中已知的。 參見例如Ernst 等人，Determination of Equilibrium Dissociation Constants, Therapeutic Monoclonal Antibodies(Wiley & Sons編，2009)。此等方法包括但不限固相結合分析(例如ELISA分析)、免疫沈澱、表面電漿子共振(SPR，例如Biacore™ (GE Healthcare, Piscataway, NJ))、動力排除分析(例如KinExA®)、流式細胞量測術、螢光活化細胞分選(FACS)、BioLayer干涉量測術(例如Octet™ (FortéBio, Inc., Menlo Park, CA))及西方墨點分析。SPR技術綜述於例如Hahnfeld 等人，Determination of Kinetic Data Using SPR Biosensors, Molecular Diagnosis of Infectious Diseases(2004)中。在典型SPR實驗中，將一種相互作用劑(目標或靶向劑)固定於流動槽中之SPR活性的鍍金玻璃載片上，且引入含有另一相互作用劑之樣本以流動通過表面。當具有給定波長之光照射在表面上時，金之光學反射率之變化指示結合及結合動力學。在一些實施例中，動力排除分析用於測定親和力。此技術描述於例如Darling等人， Assay and Drug Development Technologies第2卷，編號6 647-657 (2004)中。在一些實施例中，BioLayer干涉量測術分析用於測定親和力。此技術描述於例如Wilson等人， Biochemistry and Molecular Biology Education, 38:400-407 (2010)；Dysinger等人， J. Immunol. Methods, 379:30-41 (2012)中。 IV. 治療方法 Methods for analyzing binding affinity, binding kinetics and cross-reactivity are known in the art. See, eg , Ernst et al., Determination of Equilibrium Dissociation Constants, Therapeutic Monoclonal Antibodies (eds. Wiley & Sons, 2009). Such methods include, but are not limited to, solid phase binding assays (eg, ELISA assays), immunoprecipitation, surface plasmon resonance (SPR, eg, Biacore™ (GE Healthcare, Piscataway, NJ)), kinetic exclusion assays (eg, KinExA®), Flow cytometry, fluorescence-activated cell sorting (FACS), BioLayer interferometry (eg, Octet™ (FortéBio, Inc., Menlo Park, CA)), and Western blot analysis. SPR techniques are reviewed, for example, in Hahnfeld et al. , Determination of Kinetic Data Using SPR Biosensors, Molecular Diagnosis of Infectious Diseases (2004). In a typical SPR experiment, one interacting agent (target or targeting agent) is immobilized on an SPR-active gold-coated glass slide in a flow cell, and a sample containing the other interacting agent is introduced to flow across the surface. Changes in the optical reflectivity of gold when light of a given wavelength is impinged on the surface are indicative of binding and binding kinetics. In some embodiments, kinetic exclusion assays are used to determine affinity. This technique is described, for example, in Darling et al., Assay and Drug Development Technologies Vol. 2, No. 6 647-657 (2004). In some embodiments, BioLayer interferometry analysis is used to determine affinity. This technique is described, for example, in Wilson et al, Biochemistry and Molecular Biology Education , 38:400-407 (2010); Dysinger et al, J. Immunol. Methods , 379:30-41 (2012). IV. METHODS OF TREATMENT

在一些實施例中，提供用於治療個體之癌症的方法。在一些實施例中，該方法包含向個體投與：(1)抗體-藥物結合物(ADC)，其包含結合腫瘤相關抗原之第一抗體及細胞毒性劑，其中細胞毒性劑為微管蛋白干擾劑；以及(2)第二抗體，其結合於免疫細胞接合子，其中第二抗體包含增強與一或多種活化FcγR之結合的Fc。在一些實施例中，第二抗體之Fc增強與FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者的結合。在一些實施例中，第二抗體之Fc減弱與一或多種抑制性FcγR之結合。在一些實施例中，第二抗體之Fc減弱與FcγRIIb之結合。In some embodiments, methods for treating cancer in an individual are provided. In some embodiments, the method comprises administering to the individual: (1) an antibody-drug conjugate (ADC) comprising a primary antibody that binds a tumor-associated antigen and a cytotoxic agent, wherein the cytotoxic agent is tubulin interference and (2) a second antibody that binds to an immune cell engager, wherein the second antibody comprises an Fc that enhances binding to one or more activating FcγRs. In some embodiments, the Fc of the second antibody enhances binding to one or more of FcyRIIIa, FcyRIIa, and/or FcyRI. In some embodiments, the Fc of the second antibody reduces binding to one or more inhibitory FcγRs. In some embodiments, the Fc of the second antibody reduces binding to FcyRIIb.

在一些實施例中，一種治療癌症之方法包含向患有癌症之個體投與：(1)抗體-藥物結合物(ADC)，其中該ADC包含結合腫瘤相關抗原之第一抗體及細胞毒性劑，其中細胞毒性劑為微管蛋白干擾劑；以及(2)第二抗體，其結合免疫細胞接合子，其中第二抗體包含相對於相同同種型之野生型Fc具有增強ADCC活性之Fc。在一些實施例中，第二抗體包含相對於相同同種型之對應野生型Fc具有增強ADCC及ADCP活性的Fc。在一些實施例中，第二抗體之Fc增強與FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者的結合。在一些實施例中，第二抗體之Fc減弱與一或多種抑制性FcγR之結合。在一些實施例中，第二抗體之Fc減弱與FcγRIIb之結合。In some embodiments, a method of treating cancer comprises administering to an individual having cancer: (1) an antibody-drug conjugate (ADC), wherein the ADC comprises a first antibody that binds a tumor-associated antigen and a cytotoxic agent, wherein the cytotoxic agent is a tubulin interfering agent; and (2) a second antibody that binds an immune cell engager, wherein the second antibody comprises an Fc having enhanced ADCC activity relative to a wild-type Fc of the same isotype. In some embodiments, the second antibody comprises an Fc having enhanced ADCC and ADCP activity relative to a corresponding wild-type Fc of the same isotype. In some embodiments, the Fc of the second antibody enhances binding to one or more of FcyRIIIa, FcyRIIa, and/or FcyRI. In some embodiments, the Fc of the second antibody reduces binding to one or more inhibitory FcγRs. In some embodiments, the Fc of the second antibody reduces binding to FcyRIIb.

在各種實施例中，第二抗體為未經岩藻糖基化抗體。在各種此類實施例中，第二抗體包含於抗體組合物中，其中該組合物中之至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%之抗體未經岩藻糖基化。In various embodiments, the second antibody is an unfucosylated antibody. In various such embodiments, the second antibody is included in an antibody composition, wherein at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96% of the composition %, at least 97%, at least 98%, or at least 99% of the antibodies are not fucosylated.

在一些實施例中，第二抗體結合TIGIT。在一些實施例中，第二抗體結合CD40。在一些實施例中，第二抗體結合本文提供之免疫細胞接合子。In some embodiments, the second antibody binds TIGIT. In some embodiments, the second antibody binds CD40. In some embodiments, the second antibody binds to an immune cell adaptor provided herein.

在各種實施例中，ADC中與第一抗體結合之微管蛋白干擾劑為奧瑞他汀、微管蛋白裂解素、秋水仙鹼、長春花生物鹼、紫杉烷、克瑞托欣、類美登素或哈米特林。在一些實施例中，ADC包含MMAE或MMAF。在各種實施例中，第一抗體結合腫瘤相關抗原，諸如本文提供之腫瘤相關抗原。In various embodiments, the tubulin interfering agent in the ADC that binds to the primary antibody is auristatin, tubulin lysin, colchicine, vinca alkaloids, taxanes, critoxin, mesitoids Denso or Hamitlin. In some embodiments, the ADC comprises MMAE or MMAF. In various embodiments, the first antibody binds a tumor-associated antigen, such as a tumor-associated antigen provided herein.

本文所描述之任何ADC可與結合本文所描述之免疫細胞接合子的任何抗體組合。舉例而言，在一些實施例中，ADC為SGN-PDL1V且第二抗體為SEA-BCMA。在一些實施例中，ADC為SGN-ALPV且第二抗體為SEA-BCMA。在一些實施例中，ADC為SGN-B7H4V且第二抗體為SEA-BCMA。在一些實施例中，ADC為立伐土珠單抗維多汀且第二抗體為SEA-BCMA。在一些實施例中，ADC為SEA-CD40且第二抗體為SEA-BCMA。在一些實施例中，ADC為SEA-CD70且第二抗體為SEA-BCMA。在一些實施例中，ADC為SGN-B6A且第二抗體為SEA-BCMA。在一些實施例中，ADC為SGN-CD228A且第二抗體為SEA-BCMA。在一些實施例中，ADC為SGN-LIV1A且第二抗體為SEA-BCMA。在一些實施例中，ADC為SGN-STNV且第二抗體為SEA-BCMA。在一些實施例中，ADC為本妥昔單抗維多汀(SGN-35)且第二抗體為SEA-BCMA。在一些實施例中，ADC為恩諾單抗維多汀且第二抗體為SEA-BCMA。在一些實施例中，ADC為迪西妥單抗維多汀且第二抗體為SEA-BCMA。在一些實施例中，ADC為替索圖單抗維多汀且第二抗體為SEA-BCMA。Any of the ADCs described herein can be combined with any antibody that binds the immune cell engagers described herein. For example, in some embodiments, the ADC is SGN-PDL1V and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is SGN-ALPV and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is SGN-B7H4V and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is rivatuzumab vedotin and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is SEA-CD40 and the second antibody is SEA-BCMA. In some embodiments, the ADC is SEA-CD70 and the second antibody is SEA-BCMA. In some embodiments, the ADC is SGN-B6A and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is SGN-CD228A and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is SGN-LIV1A and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is SGN-STNV and the second antibody is SEA-BCMA. In some embodiments, the ADC is bentuximab vedotin (SGN-35) and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is ennozumab vedotin and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is dicituzumab vedotin and the secondary antibody is SEA-BCMA. In some embodiments, the ADC is tesotumumab vedotin and the secondary antibody is SEA-BCMA.

在一些實施例中，ADC為SGN-PDL1V且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-ALPV且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-B7H4V且第二抗體為SEA-CD40。在一些實施例中，ADC為立伐土珠單抗維多汀且第二抗體為SEA-CD40。在一些實施例中，ADC為SEA-CD40且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-CD70且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-B6A且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-CD228A且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-LIV1A且第二抗體為SEA-CD40。在一些實施例中，ADC為SGN-STNV且第二抗體為SEA-CD40。在一些實施例中，ADC為本妥昔單抗維多汀(SGN-35)且第二抗體為SEA-CD40。在一些實施例中，ADC為恩諾單抗維多汀且第二抗體為SEA-CD40。在一些實施例中，ADC為迪西妥單抗維多汀且第二抗體為SEA-CD40。在一些實施例中，ADC為替索圖單抗維多汀且第二抗體為SEA-CD40。In some embodiments, the ADC is SGN-PDL1V and the secondary antibody is SEA-CD40. In some embodiments, the ADC is SGN-ALPV and the secondary antibody is SEA-CD40. In some embodiments, the ADC is SGN-B7H4V and the secondary antibody is SEA-CD40. In some embodiments, the ADC is rivatuzumab vedotin and the secondary antibody is SEA-CD40. In some embodiments, the ADC is SEA-CD40 and the second antibody is SEA-CD40. In some embodiments, the ADC is SGN-CD70 and the second antibody is SEA-CD40. In some embodiments, the ADC is SGN-B6A and the secondary antibody is SEA-CD40. In some embodiments, the ADC is SGN-CD228A and the secondary antibody is SEA-CD40. In some embodiments, the ADC is SGN-LIV1A and the secondary antibody is SEA-CD40. In some embodiments, the ADC is SGN-STNV and the second antibody is SEA-CD40. In some embodiments, the ADC is bentuximab vedotin (SGN-35) and the secondary antibody is SEA-CD40. In some embodiments, the ADC is ennozumab vedotin and the secondary antibody is SEA-CD40. In some embodiments, the ADC is dicituzumab vedotin and the secondary antibody is SEA-CD40. In some embodiments, the ADC is tesotumumab vedotin and the secondary antibody is SEA-CD40.

在一些實施例中，ADC為SGN-PDL1V且第二抗體為SEA-CD70。在一些實施例中，ADC為SGN-ALPV且第二抗體為SEA-CD70。在一些實施例中，ADC為SGN-B7H4V且第二抗體為SEA-CD70。在一些實施例中，ADC為立伐土珠單抗維多汀且第二抗體為SEA-CD70。在一些實施例中，ADC為SEA-CD40且第二抗體為SEA-CD70。在一些實施例中，ADC為SEA-CD70且第二抗體為SEA-CD70。在一些實施例中，ADC為SGN-B6A且第二抗體為SEA-CD70。在一些實施例中，ADC為SGN-CD228A且第二抗體為SEA-CD70。在一些實施例中，ADC為SGN-LIV1A且第二抗體為SEA-CD70。在一些實施例中，ADC為SGN-STNV且第二抗體為SEA-CD70。在一些實施例中，ADC為本妥昔單抗維多汀(SGN-35)且第二抗體為SEA-CD70。在一些實施例中，ADC為恩諾單抗維多汀且第二抗體為SEA-CD70。在一些實施例中，ADC為迪西妥單抗維多汀且第二抗體為SEA-CD70。在一些實施例中，ADC為替索圖單抗維多汀且第二抗體為SEA-CD70。In some embodiments, the ADC is SGN-PDL1V and the secondary antibody is SEA-CD70. In some embodiments, the ADC is SGN-ALPV and the secondary antibody is SEA-CD70. In some embodiments, the ADC is SGN-B7H4V and the secondary antibody is SEA-CD70. In some embodiments, the ADC is rivatuzumab vedotin and the secondary antibody is SEA-CD70. In some embodiments, the ADC is SEA-CD40 and the second antibody is SEA-CD70. In some embodiments, the ADC is SEA-CD70 and the second antibody is SEA-CD70. In some embodiments, the ADC is SGN-B6A and the secondary antibody is SEA-CD70. In some embodiments, the ADC is SGN-CD228A and the secondary antibody is SEA-CD70. In some embodiments, the ADC is SGN-LIV1A and the secondary antibody is SEA-CD70. In some embodiments, the ADC is SGN-STNV and the second antibody is SEA-CD70. In some embodiments, the ADC is bentuximab vedotin (SGN-35) and the secondary antibody is SEA-CD70. In some embodiments, the ADC is ennozumab vedotin and the secondary antibody is SEA-CD70. In some embodiments, the ADC is dicituzumab vedotin and the secondary antibody is SEA-CD70. In some embodiments, the ADC is tesotumumab vedotin and the secondary antibody is SEA-CD70.

在一些實施例中，ADC為SGN-PDL1V且第二抗體為SEA-TGT。在一些實施例中，ADC為SGN-ALPV且第二抗體為SEA-TGT。在一些實施例中，ADC為SGN-B7H4V且第二抗體為SEA-TGT。在一些實施例中，ADC為立伐土珠單抗維多汀且第二抗體為SEA-TGT。在一些實施例中，ADC為SEA-CD40且第二抗體為SEA-TGT。在一些實施例中，ADC為SEA-CD70且第二抗體為SEA-TGT。在一些實施例中，ADC為SGN-B6A且第二抗體為SEA-TGT。在一些實施例中，ADC為SGN-CD228A且第二抗體為SEA-TGT。在一些實施例中，ADC為SGN-LIV1A且第二抗體為SEA-TGT。在一些實施例中，ADC為SGN-STNV且第二抗體為SEA-TGT。在一些實施例中，ADC為本妥昔單抗維多汀(SGN-35)且第二抗體為SEA-TGT。在一些實施例中，ADC為恩諾單抗維多汀且第二抗體為SEA-TGT。在一些實施例中，ADC為迪西妥單抗維多汀且第二抗體為SEA-TGT。在一些實施例中，ADC為替索圖單抗維多汀且第二抗體為SEA-TGT。In some embodiments, the ADC is SGN-PDL1V and the secondary antibody is SEA-TGT. In some embodiments, the ADC is SGN-ALPV and the secondary antibody is SEA-TGT. In some embodiments, the ADC is SGN-B7H4V and the secondary antibody is SEA-TGT. In some embodiments, the ADC is rivatuzumab vedotin and the secondary antibody is SEA-TGT. In some embodiments, the ADC is SEA-CD40 and the second antibody is SEA-TGT. In some embodiments, the ADC is SEA-CD70 and the second antibody is SEA-TGT. In some embodiments, the ADC is SGN-B6A and the secondary antibody is SEA-TGT. In some embodiments, the ADC is SGN-CD228A and the secondary antibody is SEA-TGT. In some embodiments, the ADC is SGN-LIV1A and the secondary antibody is SEA-TGT. In some embodiments, the ADC is SGN-STNV and the second antibody is SEA-TGT. In some embodiments, the ADC is bentuximab vedotin (SGN-35) and the secondary antibody is SEA-TGT. In some embodiments, the ADC is ennozumab vedotin and the secondary antibody is SEA-TGT. In some embodiments, the ADC is dicituzumab vedotin and the secondary antibody is SEA-TGT. In some embodiments, the ADC is tesotumumab vedotin and the secondary antibody is SEA-TGT.

在一些實施例中，個體為人類。In some embodiments, the individual is a human.

在一些實施例中，癌症為膀胱癌、乳癌、子宮癌、子宮頸癌、卵巢癌、前列腺癌、睪丸癌、食道癌、胃腸癌、胃癌(gastric cancer)、胰臟癌、大腸直腸癌、大腸癌、腎癌、透明細胞腎癌瘤、頭頸癌、肺癌、肺腺癌、胃癌(stomach cancer)、生殖細胞癌、骨癌、肝癌、甲狀腺癌、皮膚癌、黑素瘤、中樞神經系統贅瘤、間皮瘤、淋巴瘤、白血病、慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤、霍奇金氏淋巴瘤(Hodgkin lymphoma)、骨髓瘤或肉瘤。在一些實施例中，癌症選自胃癌、睪丸癌、胰臟癌、肺腺癌、膀胱癌、頭頸癌、前列腺癌、乳癌、間皮瘤及透明細胞腎癌瘤。在一些實施例中，癌症為淋巴瘤或白血病，包括但不限於急性骨髓、慢性骨髓、急性淋巴細胞或慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤、套細胞淋巴瘤、小淋巴細胞淋巴瘤、原發性縱隔大B細胞淋巴瘤、脾邊緣區B細胞淋巴瘤或結外邊緣區B細胞淋巴瘤。在一些實施例中，癌症係選自慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤及霍奇金氏淋巴瘤。在一些實施例中，乳癌為轉移性癌症。 In some embodiments, the cancer is bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, gastric cancer, pancreatic cancer, colorectal cancer, colorectal cancer Carcinoma, kidney cancer, clear cell renal carcinoma, head and neck cancer, lung cancer, lung adenocarcinoma, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, melanoma, central nervous system neoplasms , mesothelioma, lymphoma, leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, Hodgkin lymphoma, myeloma or sarcoma. In some embodiments, the cancer is selected from gastric cancer, testicular cancer, pancreatic cancer, lung adenocarcinoma, bladder cancer, head and neck cancer, prostate cancer, breast cancer, mesothelioma, and clear cell renal carcinoma. In some embodiments, the cancer is lymphoma or leukemia, including but not limited to acute myeloid, chronic myeloid, acute lymphocytic or chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, mantle cell lymphoma , small lymphocytic lymphoma, primary mediastinal large B-cell lymphoma, splenic marginal zone B-cell lymphoma, or extranodal marginal zone B-cell lymphoma. In some embodiments, the cancer line is selected from chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, and Hodgkin's lymphoma. In some embodiments, the breast cancer is metastatic cancer.

在一些實施例中，癌症為具有高腫瘤突變負荷之癌症，因而癌症具有更多驅動T細胞反應之抗原。因此，在一些實施例中，癌症為高突變負荷癌症，諸如肺癌、黑素瘤、膀胱癌或胃癌。在一些實施例中，癌症具有微衛星不穩定性。 In some embodiments, the cancer is one with a high tumor mutational burden, and thus the cancer has more antigens that drive T cell responses. Thus, in some embodiments, the cancer is a high mutational burden cancer, such as lung cancer, melanoma, bladder cancer, or gastric cancer. In some embodiments, the cancer has microsatellite instability.

在各種實施例中，第二抗體耗盡T調節性(Treg)細胞，活化抗原呈遞細胞(APC)，增強CD8 T細胞反應，上調共刺激受體及/或促進釋放免疫活化細胞介素(諸如CXCL10及/或IFNγ)。在一些實施例中，第二抗體促進免疫活化細胞介素釋放的程度大於免疫抑制細胞介素(諸如IL10及/或MDC)。 In various embodiments, the secondary antibody depletes T regulatory (Treg) cells, activates antigen presenting cells (APCs), enhances CD8 T cell responses, upregulates costimulatory receptors, and/or promotes the release of immune activating interferons such as CXCL10 and/or IFNγ). In some embodiments, the second antibody promotes the release of immune-activating interleukins to a greater extent than immune-suppressing interleukins (such as IL10 and/or MDC).

ADC與第二抗體可經同時或依序投與。對於依序投藥，可在第二抗體之第一劑量之前投與ADC之第一劑量，或可在ADC之前投與第二抗體之第一劑量。對於同時投藥，在一些實施例中，ADC與第二抗體可以單獨醫藥組合物形式或以同一醫藥組合物形式投與。 The ADC and the secondary antibody can be administered simultaneously or sequentially. For sequential administration, the first dose of the ADC may be administered before the first dose of the second antibody, or the first dose of the second antibody may be administered before the ADC. For simultaneous administration, in some embodiments, the ADC and the second antibody can be administered in separate pharmaceutical compositions or in the same pharmaceutical composition.

在一些實施例中，以治療有效量或劑量投與治療劑。可使用約0.01 mg/kg至約500 mg/kg、或約0.1 mg/kg至約200 mg/kg、或約1 mg/kg至約100 mg/kg、或約10 mg/kg至約50 mg/kg之日劑量範圍。然而，劑量可根據若干因素變化，包括所選投藥途徑、組合物之調配物、患者反應、病狀之嚴重程度、個體之重量及處方醫師之判斷。視個別患者需要，劑量可隨時間推移而增加或減少。在某些情況下，最初給予患者低劑量，接著增加至對患者可耐受之有效劑量。有效量之測定完全在熟習此項技術者之能力範圍內。In some embodiments, the therapeutic agent is administered in a therapeutically effective amount or dose. From about 0.01 mg/kg to about 500 mg/kg, or from about 0.1 mg/kg to about 200 mg/kg, or from about 1 mg/kg to about 100 mg/kg, or from about 10 mg/kg to about 50 mg /kg daily dose range. However, the dosage may vary depending on several factors, including the route of administration chosen, the formulation of the composition, the patient's response, the severity of the condition, the weight of the individual, and the judgment of the prescribing physician. The dose may be increased or decreased over time as required by the individual patient. In certain instances, the patient is initially given a low dose, which is then increased to an effective dose tolerable to the patient. Determination of an effective amount is well within the ability of those skilled in the art.

在一些實施例中，藉由本文所描述之特定組合療法觀察到的增強的活性與對應的單一療法治療相比具有某些益處。舉例而言，在一些實施例中，組合地投與ADC及第二抗體具有與以單藥療法形式投與ADC或第二抗體時相當的毒性概況。在一些實施例中，ADC及/或第二抗體在組合給藥時之有效劑量小在以單藥療法形式投與時之有效劑量。在一些實施例中，與對應單藥療法治療相比，ADC與第二抗體之組合投藥提供較長反應持續時間。在一些實施例中，與對應單藥療法相比，ADC與第二抗體之組合投藥產生較長無進展存活期。在一些實施例中，ADC及第二抗體之投藥可用於治療在單獨用任一藥劑進行之單藥療法治療後復發的復發性癌症。 In some embodiments, the enhanced activity observed with a particular combination therapy described herein has certain benefits compared to the corresponding monotherapy treatment. For example, in some embodiments, administering the ADC and the second antibody in combination has a comparable toxicity profile as when the ADC or the second antibody is administered as a monotherapy. In some embodiments, the effective dose of the ADC and/or the second antibody when administered in combination is less than when administered as a monotherapy. In some embodiments, administration of the ADC in combination with the second antibody provides a longer duration of response than the corresponding monotherapy treatment. In some embodiments, administration of the ADC in combination with the second antibody results in longer progression-free survival compared to the corresponding monotherapy. In some embodiments, administration of an ADC and a second antibody can be used to treat recurrent cancers that recur after monotherapy treatment with either agent alone.

醫藥組合物之投藥途徑可為經口、腹膜內、經皮、皮下、靜脈內、肌肉內、吸入、局部、病灶內、經直腸、支氣管內、經鼻、經黏膜、經腸道、經眼或經耳遞送，或此項技術中已知之任何其他方法。在一些實施例中，一或多種治療劑經口、靜脈內或腹膜內投與。The route of administration of the pharmaceutical composition can be oral, intraperitoneal, transdermal, subcutaneous, intravenous, intramuscular, inhalation, topical, intralesional, rectal, intrabronchial, nasal, transmucosal, enteral, ocular Or otic delivery, or any other method known in the art. In some embodiments, the one or more therapeutic agents are administered orally, intravenously, or intraperitoneally.

共投與治療劑可一起或單獨地、同時或在不同時間投與。當投與時，治療劑可視需要獨立地每天投與一次、兩次、三次、四次或更多次或更少次。在一些實施例中，所投與治療劑經每天投與一次。在一些實施例中，所投與治療劑與例如混雜物以相同時間或次數投與。在一些實施例中，治療劑中之一或多者以持續釋放調配物形式投與。Co-administration The therapeutic agents can be administered together or separately, simultaneously or at different times. When administered, the therapeutic agent may be administered independently one, two, three, four or more or fewer times per day as desired. In some embodiments, the administered therapeutic agent is administered once a day. In some embodiments, the therapeutic agent is administered at the same time or number of times as, eg, the admixture. In some embodiments, one or more of the therapeutic agents are administered in a sustained release formulation.

在一些實施例中，治療劑經同時投與。在一些實施例中，治療劑經依序投與。舉例而言，在一些實施例中，第一治療劑係在投與第二治療劑之前例如約1、2、3、4、5、6、7、8、9、10、15、20、25、30、40、50、60、70、80、90、100天或更多天投與。In some embodiments, the therapeutic agents are administered simultaneously. In some embodiments, the therapeutic agents are administered sequentially. For example, in some embodiments, the first therapeutic agent is administered, eg, about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25 before administration of the second therapeutic agent , 30, 40, 50, 60, 70, 80, 90, 100 or more days to cast.

在一些實施例中，向個體投與本文中提供之治療持續例如至少30、40、50、60、70、80、90、100、150、200、250、300、350天或更久的延長的時間段。 V.  組合物及套組 In some embodiments, the treatment provided herein is administered to an individual for an extended period of, eg, at least 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350 days or more period. V. Compositions and Kits

在另一態樣中，提供用於治療或預防個體之癌症的組合物及套組。 醫藥組合物 In another aspect, compositions and kits for treating or preventing cancer in an individual are provided. pharmaceutical composition

在一些實施例中，提供用於本發明方法之醫藥組合物。在一些實施例中，以第一醫藥組合物形式投與ADC，且以第二醫藥組合物形式投與結合免疫細胞接合子之抗體。在一些實施例中，以單一醫藥組合物形式投與ADC及結合免疫細胞接合子之抗體。In some embodiments, pharmaceutical compositions for use in the methods of the present invention are provided. In some embodiments, the ADC is administered in a first pharmaceutical composition, and the antibody that binds the immune cell engager is administered in a second pharmaceutical composition. In some embodiments, the ADC and the antibody that binds the immune cell engager are administered in a single pharmaceutical composition.

用於製備用於本發明之調配物的指導發現於例如 Remington: The Science and Practice of Pharmacy, 第21版, 2006, 見上文； Martindale: The Complete Drug Reference, Sweetman, 2005, London: Pharmaceutical Press；Niazi, Handbook of Pharmaceutical Manufacturing Formulations, 2004, CRC Press；以及Gibson, Pharmaceutical Preformulation and Formulation: A Practical Guide from Candidate Drug Selection to Commercial Dosage Form, 2001, Interpharm Press中，此等參考特此以引用之方式併入本文中。本文中描述之醫藥組合物可以熟習此項技術者已知之方式，亦即藉助於習知混合、溶解、粒化、糖衣藥丸製造、乳化、囊封、包覆或凍乾過程製造。以下方法及賦形劑僅為例示性的且決不為限制性的。 Guidance for preparing formulations for use in the present invention is found, for example, in Remington: The Science and Practice of Pharmacy , 21st Ed., 2006, supra; Martindale: The Complete Drug Reference , Sweetman, 2005, London: Pharmaceutical Press; Niazi, Handbook of Pharmaceutical Manufacturing Formulations , 2004, CRC Press; and Gibson, Pharmaceutical Preformulation and Formulation: A Practical Guide from Candidate Drug Selection to Commercial Dosage Form , 2001, Interpharm Press, these references are hereby incorporated by reference middle. The pharmaceutical compositions described herein can be manufactured in a manner known to those skilled in the art, ie by means of conventional mixing, dissolving, granulating, dragee-making, emulsifying, encapsulating, coating or lyophilizing processes. The following methods and excipients are exemplary only and in no way limiting.

在一些實施例中，製備一或多種治療劑以用於在持續釋放、控制釋放、延長釋放、定時釋放或延遲釋放調配物中，例如在含有治療劑之固體疏水性聚合物之半滲透基質中遞送。已確立各種類型之持續釋放物質且為熟習此項技術者所熟知。當前延長釋放調配物包括膜衣錠劑、多微粒或丸粒系統、使用親水性或親油性物質之基質技術及具有成孔賦形劑之基於蠟之錠劑(參見例如Huang等人， Drug Dev. Ind. Pharm.29:79 (2003)；Pearnchob等人， Drug Dev. Ind. Pharm.29:925 (2003)；Maggi,等人， Eur. J. Pharm. Biopharm.55:99 (2003)；Khanvilkar,等人， Drug Dev. Ind. Pharm.228:601 (2002)；以及Schmidt等人， Int. J. Pharm.216:9 (2001))。視其設計而定，持續釋放遞送系統可在數小時或數天過程中釋放化合物，例如經4、6、8、10、12、16、20、24小時或更長。通常，持續釋放調配物可使用天然存在的或合成聚合物來製備，該等天然存在的或合成聚合物例如聚合乙烯基吡咯啶酮，諸如聚乙烯吡咯啶酮(PVP)；羧基乙烯基親水性聚合物；疏水性及/或親水性親水膠體，諸如甲基纖維素、乙基纖維素、羥基丙基纖維素及羥基丙基甲基纖維素；以及羧基聚亞甲基。 In some embodiments, one or more therapeutic agents are prepared for use in sustained release, controlled release, extended release, timed release, or delayed release formulations, such as in a semipermeable matrix of a solid hydrophobic polymer containing the therapeutic agent deliver. Various types of sustained release substances have been established and are well known to those skilled in the art. Current extended release formulations include film-coated tablets, multiparticulate or pellet systems, matrix technologies using hydrophilic or lipophilic substances, and wax-based tablets with pore-forming excipients (see, eg, Huang et al., Drug Dev 29:79 (2003); Pearchob et al., Drug Dev. Ind. Pharm. 29 :925 (2003); Maggi, et al., Eur. J. Pharm. Biopharm. 55:99 (2003); Khanvilkar, et al, Drug Dev. Ind. Pharm. 228:601 (2002); and Schmidt et al, Int. J. Pharm. 216:9 (2001)). Depending on its design, sustained release delivery systems can release the compound over the course of hours or days, eg, over 4, 6, 8, 10, 12, 16, 20, 24 hours, or longer. In general, sustained release formulations can be prepared using naturally occurring or synthetic polymers such as polymeric vinylpyrrolidone, such as polyvinylpyrrolidone (PVP); carboxyvinyl hydrophilic polymers; hydrophobic and/or hydrophilic hydrocolloids, such as methylcellulose, ethylcellulose, hydroxypropylcellulose, and hydroxypropylmethylcellulose; and carboxypolymethylene.

對於經口投藥，治療劑可藉由與此項技術中熟知之醫藥學上可接受之載劑組合而容易地調配。此類載劑使得化合物能夠調配成用於由待治療之患者經口攝取之錠劑、丸劑、糖衣藥丸、膠囊、乳液、親脂性及親水性懸浮液、液體、凝膠、糖漿、漿液、懸浮液及其類似物。經口使用之醫藥製劑可藉由將化合物與固體賦形劑混合，視情況研磨所得混合物，且必要時在添加適合助劑之後加工顆粒混合物以得到錠劑或糖衣藥丸芯來獲得。適合之賦形劑包括例如填充劑，諸如糖，包括乳糖、蔗糖、甘露糖醇或山梨糖醇；纖維素製劑，諸如玉米澱粉、小麥澱粉、大米澱粉、馬鈴薯澱粉、明膠、黃蓍膠、甲基纖維素、羥丙基甲基纖維素、羧甲基纖維素鈉及/或聚乙烯吡咯啶酮(PVP)。視需要，可添加崩解劑，諸如交聯聚乙烯吡咯啶酮、瓊脂或褐藻酸或其鹽，諸如褐藻酸鈉。For oral administration, the therapeutic agent can be formulated readily by combining with pharmaceutically acceptable carriers well known in the art. Such carriers enable the compounds to be formulated for oral ingestion by the patient to be treated as lozenges, pills, dragees, capsules, emulsions, lipophilic and hydrophilic suspensions, liquids, gels, syrups, slurries, suspensions liquid and the like. Pharmaceutical preparations for oral use can be obtained by mixing the compound with a solid excipient, grinding the resulting mixture, as appropriate, and processing the mixture of granules, after adding suitable auxiliaries, if necessary, to obtain dragees or dragee cores. Suitable excipients include, for example, fillers such as sugars, including lactose, sucrose, mannitol or sorbitol; cellulosic preparations such as corn starch, wheat starch, rice starch, potato starch, gelatin, tragacanth, methyl methacrylate cellulose, hydroxypropyl methylcellulose, sodium carboxymethylcellulose and/or polyvinylpyrrolidone (PVP). If desired, a disintegrant such as cross-linked polyvinylpyrrolidone, agar or alginic acid or a salt thereof such as sodium alginate may be added.

治療劑可經調配用於藉由注射，例如藉由彈丸注射或連續輸注來進行非經腸投藥。對於注射，可藉由將一或多種化合物溶解、懸浮或乳化於水性或非水溶劑，諸如植物油或其他類似油、合成脂族酸甘油酯、高級脂族酸之酯或丙二醇中；且視需要，伴以習知添加劑，諸如增溶劑、等張劑、懸浮劑、乳化劑、穩定劑及防腐劑來將該化合物調配成製劑。在一些實施例中，化合物可以水溶液形式調配，較佳以生理相容緩衝液，諸如漢克氏溶液(Hanks's solution)、林格氏溶液(Ringer's solution)或生理鹽水緩衝液形式調配。注射用調配物可呈單位劑型，例如以安瓿或多劑量容器形式，其中添加有防腐劑。組合物可採用諸如於油性或水性媒劑中之懸浮液、溶液或乳液之形式，且可含有諸如懸浮劑、穩定劑及/或分散劑之調配劑。The therapeutic agent can be formulated for parenteral administration by injection, eg, by bolus injection or continuous infusion. For injection, by dissolving, suspending or emulsifying one or more compounds in aqueous or non-aqueous solvents, such as vegetable oils or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids, or propylene glycol; and as required , with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifiers, stabilizers and preservatives to formulate the compounds into formulations. In some embodiments, the compounds can be formulated in aqueous solutions, preferably in physiologically compatible buffers such as Hanks's solution, Ringer's solution, or physiological saline buffer. Formulations for injection may be presented in unit dosage form, eg, in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as suspending, stabilizing and/or dispersing agents.

治療劑可藉由經黏膜或經皮方式全身性投與。對於經黏膜或經皮投藥，在調配物中使用適於滲透障壁的滲透劑。對於局部投藥，藥劑經調配成軟膏、乳膏、油膏、散劑及凝膠。在一個實施例中，經皮遞送劑可為DMSO。經皮遞送系統可包括例如貼劑。對於經黏膜投藥，在調配物中使用適於滲透障壁的滲透劑。此類滲透劑通常為此項技術中已知的。例示性經皮遞送調配物包括描述於美國專利第6,589,549號；第6,544,548號；第6,517,864號；第6,512,010號；第6,465,006號；第6,379,696號；第6,312,717號及第6,310,177號中之彼等調配物，該等專利其中之每一者特此以引用之方式併入本文中。Therapeutic agents can be administered systemically by transmucosal or transdermal means. For transmucosal or transdermal administration, penetrants suitable for penetrating barriers are used in the formulation. For topical administration, the agents are formulated into ointments, creams, salves, powders and gels. In one embodiment, the transdermal delivery agent may be DMSO. Transdermal delivery systems can include, for example, patches. For transmucosal administration, penetrants suitable for penetrating barriers are used in the formulation. Such penetrants are generally known in the art. Exemplary transdermal delivery formulations include those described in US Patent Nos. 6,589,549; 6,544,548; 6,517,864; 6,512,010; 6,465,006; 6,379,696; Each of these patents is hereby incorporated herein by reference.

在一些實施例中，醫藥組合物包含可接受之載劑及/或賦形劑。醫藥學上可接受之載劑包括生理相容且較佳地不會干擾或另外抑制治療劑之活性的任何溶劑、分散介質或包衣。在一些實施例中，載劑適用於靜脈內、肌肉內、經口、腹膜內、經皮、局部或皮下投藥。醫藥學上可接受之載劑可含有一或多種生理學上可接受之化合物，其用於例如使組合物穩定或增加或減少活性劑吸收。生理學上可接受之化合物可包括例如碳水化合物，諸如葡萄糖、蔗糖、聚葡萄糖；抗氧化劑，諸如抗壞血酸或麩胱甘肽；螯合劑；低分子量蛋白質；減少活性劑之清除或水解的組合物或賦形劑或其他穩定劑及/或緩衝劑。其他醫藥學上可接受之載劑及其調配物為熟知的且一般描述於例如 Remington: The Science and Practice of Pharmacy, 第21版, Philadelphia, PA. Lippincott Williams & Wilkins, 2005中。各種醫藥學上可接受之賦形劑為此項技術中熟知的且可發現於例如Handbook of Pharmaceutical Excipients (第5版, Ed. Rowe等人，Pharmaceutical Press, Washington, D.C.)中 In some embodiments, the pharmaceutical composition includes acceptable carriers and/or excipients. Pharmaceutically acceptable carriers include any solvent, dispersion medium or coating that is physiologically compatible and preferably does not interfere with or otherwise inhibit the activity of the therapeutic agent. In some embodiments, the carrier is suitable for intravenous, intramuscular, oral, intraperitoneal, transdermal, topical or subcutaneous administration. A pharmaceutically acceptable carrier may contain one or more physiologically acceptable compounds, which are used, for example, to stabilize the composition or to increase or decrease absorption of the active agent. Physiologically acceptable compounds may include, for example, carbohydrates, such as glucose, sucrose, polydextrose; antioxidants, such as ascorbic acid or glutathione; chelating agents; low molecular weight proteins; compositions that reduce clearance or hydrolysis of active agents; or Excipients or other stabilizers and/or buffers. Other pharmaceutically acceptable carriers and their formulations are well known and generally described, eg, in Remington: The Science and Practice of Pharmacy , 21st Ed., Philadelphia, PA. Lippincott Williams & Wilkins, 2005. Various pharmaceutically acceptable excipients are well known in the art and can be found, for example, in the Handbook of Pharmaceutical Excipients (5th Edition, Ed. Rowe et al., Pharmaceutical Press, Washington, DC)

本發明之醫藥組合物之劑量及所需濃度可視所設想之特定用途而變化。測定適合的劑量或投藥途徑完全在熟習此項技術者之技能內。本文中亦描述適合之劑量。 套組 The dosages and desired concentrations of the pharmaceutical compositions of the present invention may vary depending upon the particular use contemplated. Determination of appropriate doses or routes of administration is well within the skill of those skilled in the art. Suitable dosages are also described herein. set

在一些實施例中，提供用於治療患有癌症之個體之套組。在一些實施例中，該套組包含： 抗體-藥物結合物，其包含與微管蛋白干擾劑結合之第一抗體，如本文所提供；以及 第二抗體，其結合免疫細胞接合子，如本文所提供。 In some embodiments, kits are provided for treating individuals with cancer. In some embodiments, the kit includes: An antibody-drug conjugate comprising a first antibody bound to a tubulin interfering agent, as provided herein; and The secondary antibody, which binds to the immune cell engager, is provided herein.

在一些實施例中，套組可以進一步包含含有用於實踐本發明方法之說明(亦即協定)的說明材料(例如使用治療癌症之套組之說明書)。雖然說明材料典型地包含書面或印刷材料，但其不限於此。本發明涵蓋能夠儲存此等說明書且將其傳達至終端使用者之任何媒介。此類媒體包括但不限於電子儲存媒體(例如磁碟、磁帶、藥筒、晶片)、光學媒體(例如，CD ROM)及其類似媒體。此類媒體可包括提供此類說明材料之網際網路位點的位址。 VI. 實例 In some embodiments, the kit may further comprise instructional material (eg, instructions for use of the kit for treating cancer) containing instructions (ie, protocols) for practicing the methods of the invention. While the instructional material typically includes written or printed material, it is not so limited. The present invention encompasses any medium capable of storing such instructions and communicating them to end users. Such media include, but are not limited to, electronic storage media (eg, magnetic disks, tapes, cartridges, wafers), optical media (eg, CD ROMs), and the like. Such media may include addresses to Internet sites that provide such instructional materials. VI. Examples

下文所論述之實例僅意欲例示本發明，且不應視為以任何方式限制本發明。該等實例不意欲表示以下實驗為所執行之所有實驗或唯一實驗。已努力確保關於所使用之數量(例如量、溫度等)的準確性，但應當考慮一些實驗性誤差及偏差。除非另有指示，否則份數為重量份，分子量為平均分子量，溫度係以攝氏度計，且壓力為大氣壓或接近大氣壓。 實例 1 ： 非直接化學治療劑損害 T 細胞 反應 1.1 物質與方法 The examples discussed below are intended to illustrate the invention only, and should not be construed as limiting the invention in any way. These examples are not intended to represent that the experiments below are all or the only experiments performed. Efforts have been made to ensure accuracy with respect to quantities used (eg, amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric. Example 1 : Indirect chemotherapeutic agents impair T cell responses 1.1 Substances and methods

使用塗佈CD3/CD28之珠粒誘導人類原代T細胞進行增殖。將20,000個經羧基螢光素二乙酸酯丁二醯亞胺基酯(CSFE)標記之富集CD3+ T細胞與抗CD3 CD28珠粒(1個珠粒/4個T細胞)+10 ng/mL IL-2一起培育4天。細胞用LIVE/DEAD Fixable Dead Cell Stain (ThermoFisher)染色且經由流式細胞量測術對活細胞進行計數。 1.2 結果 Human primary T cells were induced to proliferate using CD3/CD28-coated beads. 20,000 enriched CD3+ T cells labeled with carboxyluciferin diacetate butadiimide (CSFE) were mixed with anti-CD3 CD28 beads (1 bead/4 T cells) + 10 ng/ mL IL-2 was incubated together for 4 days. Cells were stained with LIVE/DEAD Fixable Dead Cell Stain (ThermoFisher) and viable cells were counted via flow cytometry. 1.2 Results

如圖1中所展示，所測試之所有單一免費代理化學治療劑顯著減少原代人類T細胞增殖。此等資料表明，全身性暴露於化學治療劑可能限制患者之T細胞介導之活性，包括來自免疫腫瘤學藥劑(例如結合免疫細胞接合子之抗體)之反應。 實例 2 ： 維多汀 ADC 不會抑制 T 細胞增殖， 即使直接遞送至 T 細胞 (CD30+ CD8 T 細胞 之 BV (SGN-35) 處理 ) 2.1 物質與方法 As shown in Figure 1, all single free agent chemotherapeutics tested significantly reduced primary human T cell proliferation. These data suggest that systemic exposure to chemotherapeutic agents may limit T cell-mediated activity in patients, including responses from immuno-oncology agents such as antibodies that bind to immune cell engagers. Example 2 : Verdotin ADC does not inhibit T cell proliferation, even when delivered directly to T cells ( BV (SGN-35) treatment of CD30+ CD8 T cells ) 2.1 Materials and Methods

人類原代CD8 T細胞經CSFE標記且用抗CD3-CD28珠粒(1珠粒/4個T細胞)+10 ng/mL IL-2誘導以進行增殖，持續4天。在活化期間，T細胞之表面上之CD30經上調。將CD30+ CD8 T細胞用CD30導引之vc-MMAE (本妥昔單抗維多汀；BV；SGN-35)或同型對照處理。細胞用LIVE/DEAD Fixable Dead Cell Stain (ThermoFisher)染色且經由流式細胞量測術對活細胞進行計數。 2.2 結果 Human primary CD8 T cells were CSFE labeled and induced to proliferate with anti-CD3-CD28 beads (1 bead/4 T cells) + 10 ng/mL IL-2 for 4 days. During activation, CD30 on the surface of T cells is upregulated. CD30+ CD8 T cells were treated with CD30-directed vc-MMAE (Bentuximab Verdotin; BV; SGN-35) or an isotype control. Cells were stained with LIVE/DEAD Fixable Dead Cell Stain (ThermoFisher) and viable cells were counted via flow cytometry. 2.2 Results

如圖2中所展示，原代人類CD30+ CD8 T細胞之細胞增殖未藉由用BV進行處理而顯著改變。此等資料表明全身性暴露於維多汀ADC，甚至直接靶向CD8 T細胞，仍不會影響CD8介導之抗腫瘤反應。 實例 3 ： 對於維多汀 ADC ， 內質網應激誘導係優異的 3.1 物質與方法 As shown in Figure 2, cell proliferation of primary human CD30+ CD8 T cells was not significantly altered by treatment with BV. These data suggest that systemic exposure to vedotin ADCs, even directly targeting CD8 T cells, does not affect CD8-mediated antitumor responses. Example 3 : Excellent 3.1 Substances and Methods for Inducible ER Stress for Vidotin ADCs

誘導內質網(ER)應激為用於引發免疫原性細胞反應之第一且所需步驟中之一者(圖3B)。MIA-PaCa-2胰臟癌細胞株用與不同有效負載(包括維多汀(MMAE)、恩他新(DM1)、依昔替康DS-8201 (Ex)以及游離微管穩定劑太平洋紫杉醇)結合之ADC以在此系統中誘導細胞死亡之IC50濃度處理。在處理36或48小時之後，採集細胞用於西方墨點分析及藉由西方墨點評估上游ER應激標記物pJNK (圖3B)。 3.1.1 西方墨點 Induction of endoplasmic reticulum (ER) stress is one of the first and required steps for eliciting an immunogenic cellular response (Figure 3B). MIA-PaCa-2 pancreatic cancer cell lines were used with different payloads (including vedotin (MMAE), entaxine (DM1), xinotecan DS-8201 (Ex), and the free microtubule stabilizer paclitaxel) Bound ADCs were treated at IC50 concentrations that induced cell death in this system. After 36 or 48 hours of treatment, cells were harvested for western blot analysis and assessment of the upstream ER stress marker pJNK by western blot (Figure 3B). 3.1.1 Western Ink Dots

將處理細胞以14,000至16,000 rpm離心10 min且儲存於-20℃。將細胞沈澱物再懸浮於4X BOLT™ LDS樣本緩衝液(Thermo Fisher目錄號B0007)中且藉由在95℃下加熱5至10分鐘裂解，以產生細胞裂解物。樣本裂解物在MOPS緩衝液中在140 V下在Bis-Tris 4至12%梯度凝膠上操作1小時40分鐘。接著使用iBlot2將Bis-Tris凝膠轉移至硝化纖維素膜上。在1X TBS中洗滌膜一次且在Licor阻斷緩衝液中在4℃下培育隔夜。在1X TBS-T中洗滌薄膜四次，每次持續5至10 min。在使用84 µm解析度及自動強度之Licor Odyssey系統上產生影像。 3.1.2 CHOP 螢光素酶誘導分析 Treated cells were centrifuged at 14,000 to 16,000 rpm for 10 min and stored at -20°C. Cell pellets were resuspended in 4X BOLT™ LDS sample buffer (Thermo Fisher Cat# B0007) and lysed by heating at 95°C for 5 to 10 minutes to generate cell lysates. Sample lysates were run on a Bis-Tris 4 to 12% gradient gel at 140 V in MOPS buffer for 1 hour and 40 minutes. The Bis-Tris gel was then transferred to a nitrocellulose membrane using iBlot2. Membranes were washed once in IX TBS and incubated in Licor blocking buffer overnight at 4°C. Wash the membrane four times in 1X TBS-T for 5 to 10 min each. Images were produced on a Licor Odyssey system using 84 µm resolution and automatic intensity. 3.1.2 CHOP luciferase induction assay

使用經CHOP驅動螢光素酶報導體細胞株轉導之MIA-PaCa-2細胞執行ER應激誘導之下游路徑的評估。CHOP為ER應激反應級聯中之最後一步且其表現量藉由ER應激增加。臨床發展中之若干ADC有效負載(圖3A)用於評估。使用針對CHOP活性之報導體系統根據製造商說明書(Bright-Glo™ Luciferase Assay System, Promega)來量測CHOP誘導。簡言之，將100,000個細胞/孔接種於96孔平底透明培養盤中(等分試樣，150微升/孔)。將200 µL培養基等分至培養盤之外部孔以提供圍繞細胞孔之培養基「覆層」。在24、48及72小時，自培育箱移出培養盤且使其達至室溫。自該等孔移除100 µL培養基。將100 µL BrightGlo試劑添加至各孔。在讀取之前，搖晃培養盤至少兩分鐘。使用Envision CTG 96孔標準協定以讀取培養盤。 3.2 結果 Assessment of downstream pathways induced by ER stress was performed using MIA-PaCa-2 cells transduced with a CHOP-driven luciferase reporter somatic cell line. CHOP is the last step in the ER stress response cascade and its expression is increased by ER stress. Several ADC payloads in clinical development (FIG. 3A) were used for evaluation. CHOP induction was measured using a reporter system for CHOP activity according to the manufacturer's instructions (Bright-Glo™ Luciferase Assay System, Promega). Briefly, 100,000 cells/well were seeded in 96-well flat bottom clear culture dishes (aliquot, 150 microliters/well). Aliquot 200 µL of medium into the outer wells of the culture dish to provide a medium "coating" around the cell wells. At 24, 48 and 72 hours, the plates were removed from the incubator and allowed to come to room temperature. Remove 100 µL of medium from the wells. Add 100 µL BrightGlo Reagent to each well. Shake the plate for at least two minutes before reading. Plates were read using an Envision CTG 96-well standard protocol. 3.2 Results

如圖3C至圖3E中所展示，基於奧瑞他汀之ADC (MMAE-ADC或MMAF-ADC)處理為發現以誘導所測試之不同ADC有效負載之早期ER應激反應pJNK信號的唯一情況。ER應激誘導與微管破壞相關聯，此係由於ER需要完整微管擴增及收縮以適應細胞之蛋白質轉譯需要。MMAE作為微管蛋白干擾劑誘導此ER應激之能力經例示於所示資料中。 As shown in Figures 3C-3E, auristatin-based ADC (MMAE-ADC or MMAF-ADC) treatment was the only case found to induce early ER stress response pJNK signaling for the different ADC payloads tested. ER stress induction is associated with microtubule disruption, as the ER requires intact microtubule expansion and contraction to accommodate the protein translation needs of the cell. The ability of MMAE to induce this ER stress as a tubulin interfering agent is exemplified in the data shown.

圖3D至圖3E展現CHOP (ER應激路徑中之下游信號)由MMAE ADC顯著誘發且與其他有效負載相比，ER應激之下游路徑亦由MMAE ADC不同地驅動。 實例 4 ： 不同臨床 ADC 有效負載之 ICD 潛能 4.1 物質與方法 Figures 3D-3E demonstrate that CHOP, a downstream signal in the ER stress pathway, is significantly evoked by the MMAE ADC and that the downstream pathway of ER stress is also driven differently by the MMAE ADC compared to other payloads. Example 4 : ICD Potential of Different Clinical ADC Payloads 4.1 Materials and Methods

典型ICD標記物包括表面暴露鈣網伴護蛋白且釋放ATP及HMGB1，其伴隨ER應激反應之誘導而發生。此等分子視為危險信號且活化先天免疫細胞且增加腫瘤抗原特異性T細胞反應。MIA-PaCa-2癌細胞用IC50濃度之當前處於臨床階段的攜帶ADC之有效負載，亦即MMAE、DM1及依昔替康(Ex)處理。接著分析經處理細胞之ICD標記物誘導。順鉑用作陰性對照，此係因為其能夠驅動細胞死亡，但已知不誘導ICD。 Typical ICD markers include surface-exposed calreticulin chaperrin and release of ATP and HMGB1, which occurs with induction of the ER stress response. These molecules are seen as danger signals and activate innate immune cells and increase tumor antigen-specific T cell responses. MIA-PaCa-2 cancer cells were treated with IC50 concentrations of currently clinical stage ADC-bearing payloads, namely MMAE, DM1 and ixnotecan (Ex). Treated cells were then analyzed for ICD marker induction. Cisplatin was used as a negative control due to its ability to drive cell death but is not known to induce ICD.

在96孔盤中每孔接種100,000至150,000個細胞。使細胞達到50%至60%匯合。移除培養基且每孔細胞添加新鮮培養基。將1µg/mL 1µM 藥物添加至各孔細胞中。在24小時之後，收集250 µL (用於ATP釋放分析)或200 µL (用於HMGB1分析)培養基且轉移至經標記1.5 mL埃彭道夫管(Eppendorf tube)中。各管樣本以10,000 rpm離心1 min。將50 µL培養基轉移至96孔透明底培養盤中之孔中。將50 µL CTG添加至各孔中。搖晃培養盤1至2 min。使用Envision盤式讀取器以讀取培養盤。 Seed 100,000 to 150,000 cells per well in a 96-well plate. Bring cells to 50% to 60% confluence. The medium was removed and fresh medium was added per well of cells. Add 1 µg/mL of 1 µM drug to each well of cells. After 24 hours, 250 µL (for ATP release assay) or 200 µL (for HMGB1 assay) medium was collected and transferred to a labeled 1.5 mL Eppendorf tube. Samples from each tube were centrifuged at 10,000 rpm for 1 min. Transfer 50 µL of medium to wells in a 96-well clear bottom culture dish. Add 50 µL CTG to each well. Shake the plate for 1 to 2 min. An Envision Disc Reader was used to read the culture plates.

使用Envision盤式讀取器藉由發光強度/孔監測HMGB1釋放量。HMGB1及ATP釋放量經報導為相對於未經處理樣本之背景值的倍數變化。將所獲取的值轉化成正文檔案且使用Excel及GraphPad Prism導出並分析。 4.2 結果 HMGB1 release was monitored by luminescence intensity/well using an Envision disc reader. HMGB1 and ATP release are reported as fold changes relative to background values for untreated samples. The acquired values were converted into text files and exported and analyzed using Excel and GraphPad Prism. 4.2 Results

如圖4A中所展示，與測試之其他有效負載相比，vc-MMAE強效驅動ATP釋放。儘管HMGB1釋放與ICD之誘導相關，但當細胞開始經歷壞死且不與穩固免疫細胞接合直接相關時，亦發現其釋放。與拓樸異構酶抑制劑依昔替康(Ex)對比，用微管破壞劑vc-MMAE及DM1處理MIA-PaCa-2細胞產生穩固的HMGB1釋放(圖4B)。 實例 5 ： ADC 有效負載之免疫活化評估 5.1 物質與方法 5.1.1細胞 As shown in Figure 4A, vc-MMAE drives ATP release potently compared to other payloads tested. Although HMGB1 release was associated with induction of ICD, its release was also found when cells began to undergo necrosis and was not directly associated with robust immune cell engagement. Treatment of MIA-PaCa-2 cells with the microtubule disruptors vc-MMAE and DM1 resulted in robust HMGB1 release compared to the topoisomerase inhibitor ixitecan (Ex) (Figure 4B). Example 5 : Assessment of Immune Activation of ADC Payloads 5.1 Materials and Methods 5.1.1 Cells

如圖5A中所示，與MMAE結合之ADC破壞微管，引起ER應激反應，從而導致免疫原性細胞死亡(ICD)。死亡細胞轉而釋放免疫活化分子—損傷相關分子模式(DAMP)—諸如HSP70、HSP90、ATP、HMGB1及鈣網伴護蛋白(CRT)。此等DAMP可以結合受體，諸如LPR1/CD91、P2RX7、P2RY2、AGER、TLR2及TLR4，進而活化先天免疫系統。此活化引起例如諸如CD80、CD86、HLA-DR及CD40之蛋白質上調，單核球上之MHCII表現增加及諸如CXCL-10/IP10及IL-12之細胞介素釋放，進而引發抗腫瘤T細胞反應。此類T細胞反應可藉由PD-1/L1抑制劑進一步強化。本文中，在人類周邊血液單核細胞(PBMC)培養物中評估ICD之免疫結果。將暴露於與不同有效負載結合之ADC的癌細胞添加至PBMC。 As shown in Figure 5A, ADCs bound to MMAE disrupt microtubules, causing an ER stress response leading to immunogenic cell death (ICD). Dead cells in turn release immune-activating molecules—damage-associated molecular patterns (DAMPs)—such as HSP70, HSP90, ATP, HMGB1, and calreticulin (CRT). These DAMPs can bind to receptors such as LPR1/CD91, P2RX7, P2RY2, AGER, TLR2 and TLR4, thereby activating the innate immune system. This activation causes, for example, upregulation of proteins such as CD80, CD86, HLA-DR and CD40, increased expression of MHCII on monocytes and release of interleukins such as CXCL-10/IP10 and IL-12, which in turn elicits anti-tumor T cell responses . Such T cell responses can be further enhanced by PD-1/L1 inhibitors. Herein, the immunological outcomes of ICD were assessed in human peripheral blood mononuclear cell (PBMC) cultures. Cancer cells exposed to ADCs bound to different payloads were added to PBMCs.

暴露於EC50濃度之ADC或游離藥物18小時(在37℃，5% CO 2下)中之L540cy癌細胞經洗滌且將以10×10 6個細胞/毫升懸浮之250 ul PBMC添加至癌細胞株殺死之細胞中，保持48小時。獲取組織培養基且藉由Luminex評估量測細胞介素。 L540cy cancer cells exposed to EC50 concentrations of ADC or free drug for 18 hours (at 37°C, 5% CO2 ) were washed and 250 ul of PBMC suspended at 10 x 106 cells/ml were added to the cancer cell line Killed cells for 48 hours. Tissue culture medium was obtained and intercytokinins were measured by Luminex assessment.

針對2個獨立PBMC供體執行處理，重複三次。 5.1.2 共刺激分子表面表現 Treatment was performed on 2 independent PBMC donors in triplicate. 5.1.2 Surface representation of costimulatory molecules

處理後，將細胞沈澱物再懸浮於50 mL BD FACs緩衝液中且轉移至96孔圓底微量滴定盤。Fc受體在冰上用人類100 µg/mL Fc片段封端，保持30分鐘。在含有100 mg/mL人類純化Fc片段之BD FACs緩衝液中製備由以1:100稀釋之PE-HLA-DR (MHCII)及APC-CD14構成之主混合物。將10 µl主混合物添加至含有90 µl再懸浮細胞之各孔且在冰上培育樣本1小時。接著將細胞在預先冷卻之Eppendorf 5810R離心機以400 xg離心5分鐘。移除上清液且用200 mL BD FACs緩衝液洗滌細胞。執行兩次洗滌且使細胞再懸浮於200 mL FACs緩衝液中且在Attune流式細胞儀上分析樣本。使用FlowJo分析軟體測定HLA-DR平均螢光。 5.1.3 細胞介素產生 After treatment, the cell pellet was resuspended in 50 mL of BD FACs buffer and transferred to a 96-well round bottom microtiter plate. Fc receptors were capped with human 100 µg/mL Fc fragment for 30 min on ice. A master mix consisting of PE-HLA-DR (MHCII) and APC-CD14 diluted 1:100 was prepared in BD FACs buffer containing 100 mg/mL purified human Fc fragment. 10 µl of master mix was added to each well containing 90 µl of resuspended cells and samples were incubated on ice for 1 hour. The cells were then centrifuged at 400 xg for 5 minutes in a pre-cooled Eppendorf 5810R centrifuge. The supernatant was removed and cells were washed with 200 mL of BD FACs buffer. Two washes were performed and cells were resuspended in 200 mL of FACs buffer and samples were analyzed on an Attune flow cytometer. HLA-DR mean fluorescence was determined using FlowJo analysis software. 5.1.3 Interferon production

處理之後，PBMC/癌細胞共培養物用盤配接器在Eppendorf 5810R中以800 rpm旋轉5分鐘。移除血清或組織培養物上清液且轉移至96帶試管架且將樣本冷凍在-80℃下直至處理。在4℃下解凍經冷凍組織培養上清液及血清隔夜且使用來自Millipore之Luminex多工套組處理以用於產生細胞介素。After treatment, the PBMC/cancer cell co-cultures were spun in an Eppendorf 5810R at 800 rpm for 5 minutes using a disk adapter. Serum or tissue culture supernatant was removed and transferred to a 96-belt tube rack and samples were frozen at -80°C until processing. Frozen tissue culture supernatants and sera were thawed overnight at 4°C and processed using the Luminex multiplex kit from Millipore for interferon production.

根據製造商說明書處理組織培養上清液及血清樣本。簡言之，分析盤用200微升洗滌緩衝液/孔洗滌，隨後將25 µL標準或緩衝液、25 µL基質或樣本及25 µL多工分析珠粒添加至各孔。在4℃下在劇烈振盪下培育樣本隔夜。洗滌培養盤，且用洗滌緩衝液洗滌分析盤兩次。Tissue culture supernatants and serum samples were processed according to the manufacturer's instructions. Briefly, assay plates were washed with 200 µL of wash buffer/well, followed by the addition of 25 µL of standard or buffer, 25 µL of matrix or sample, and 25 µL of multiplexed assay beads to each well. The samples were incubated overnight at 4°C with vigorous shaking. The culture plate was washed, and the assay plate was washed twice with wash buffer.

將偵測抗體(25 µL)添加至各孔且在室溫下培育1小時。添加25 µL抗生蛋白鏈菌素-藻紅素(SA-PE)且在室溫下培育樣本30分鐘。用洗滌緩衝液洗滌培養盤兩次且用150 µL鞘液再懸浮珠粒。使用Luminex MagPix系統與Xponent軟體系統的組合分析樣本。根據標準曲線計算細胞介素含量。 5.2 結果 Detection antibody (25 µL) was added to each well and incubated for 1 hour at room temperature. 25 µL of streptavidin-phycoerythrin (SA-PE) was added and the samples were incubated at room temperature for 30 minutes. The plate was washed twice with wash buffer and the beads were resuspended with 150 µL of sheath fluid. Samples were analyzed using a combination of the Luminex MagPix system and the Xponent software system. Interferon content was calculated according to the standard curve. 5.2 Results

觀測到先天細胞活化，如藉由增大的表面活化標記物(MHCII)及發炎性細胞介素(CXCL-10/IP10)之釋放所證明(圖5B至圖5C)。先天免疫細胞在暴露於vc-MMAE處理之腫瘤細胞時變得活化。藉由vc-MMAE之免疫細胞活化比藉由其他ADC有效負載之活化更穩固(圖5B至圖5C)。 Innate cell activation was observed, as evidenced by increased release of surface activation markers (MHCII) and inflammatory interleukins (CXCL-10/IP10) (FIG. 5B-FIG. 5C). Innate immune cells become activated upon exposure to vc-MMAE-treated tumor cells. Immune cell activation by vc-MMAE was more robust than activation by other ADC payloads (Figure 5B-5C).

Vc-MMAE介導之ICD為活化針對來自死亡及正在死亡之腫瘤細胞的抗原之適應性免疫反應且允許產生穩固先天免疫細胞活化及針對特定腫瘤細胞抗原之後續細胞毒性T細胞反應的經調節細胞死亡。本文中，證實vc-MMAE殺死之癌細胞引發在吸收死亡細胞之後的表面MHCII增加及自單核球/巨噬細胞釋放先天細胞介素CXCL10 (一種強趨化性及發炎性介體)。 實例 6 ： 曲妥珠單抗主鏈上之有效負載評估 6.1 物質與方法 Vc-MMAE-mediated ICD is a regulated cell that activates adaptive immune responses against antigens from dying and dying tumor cells and allows for robust innate immune cell activation and subsequent cytotoxic T cell responses against specific tumor cell antigens die. Herein, it was demonstrated that cancer cells killed by vc-MMAE trigger increased surface MHCII and release of the innate interleukin CXCL10, a potent chemotactic and inflammatory mediator, from monocytes/macrophages following uptake of dead cells. Example 6 : Evaluation of Payload on Trastuzumab Backbone 6.1 Materials and Methods

評估攜帶各種臨床階段有效負載之曲妥珠單抗ADC結合物誘導ER應激及下游ICD標記物ATP及HMGB1之能力。所使用有效負載為DM1、MMAE及依昔替康(Ex)。 6.2 結果 Trastuzumab ADC conjugates carrying various clinical stage payloads were assessed for their ability to induce ER stress and the downstream ICD markers ATP and HMGB1. The payloads used were DM1, MMAE and Ixinotecan (Ex). 6.2 Results

得到兩種觀察結果：(1)與vcMMAE結合之曲妥珠單抗驅動最穩固的ER應激反應，其與ATP及HMGB1之誘導相關聯；以及(2)晚期細胞死亡標記物HMGB1針對其他有效負載類別似乎升高，指示繼發性壞死可能與此等有效負載類別而非老實的ICD相關聯(圖6C至圖6E)。此處之發現與使用Mia-PaCa-2細胞(圖4A至圖4B)之上文實例4中所描述之發現類似。 實例 7 ： 對於 MMAE ADC ， 早期 ER 應激標記物之誘導 (JNK 傳信活化 ) 一般為優異的 7.1 物質與方法 Two observations were made: (1) trastuzumab conjugated to vcMMAE drives the most robust ER stress response, which is associated with induction of ATP and HMGB1; and (2) the late cell death marker HMGB1 is effective against other The payload classes appeared to be elevated, indicating that secondary necrosis may be associated with these payload classes rather than honest ICD (Figures 6C-6E). The findings here are similar to those described in Example 4 above using Mia-PaCa-2 cells (Figures 4A-4B). Example 7 : Induction of early ER stress markers (JNK signaling activation ) is generally excellent for MMAE ADCs 7.1 Substances and Methods

如實例5中所描述且圖5A中所示，ICD路徑涉及各種態樣。此路徑進一步說明於圖7A中。如圖7A中所示及上文提及，處於初始階段之諸如MMAE之微管蛋白干擾劑破壞微管，進而致使ER應激及ICD。ICD轉而造成釋放免疫活化分子，諸如DAMP、ATP、HMGB1及CRT。此等分子可隨後活化能夠引發抗腫瘤T細胞反應且可誘導T細胞記憶之先天細胞。此類T細胞反應可藉由與其他免疫調節物，諸如本文所描述之免疫細胞接合子組合而進一步強化。若干以下實例報導研究結果，其展示與其他ADC有效負載相比，MMAE誘導ICD路徑之前述不同態樣之有效性。 As described in Example 5 and shown in Figure 5A, the ICD path involves various aspects. This path is further illustrated in Figure 7A. As shown in Figure 7A and mentioned above, tubulin interfering agents such as MMAE at the initial stage disrupt microtubules, resulting in ER stress and ICD. ICD in turn results in the release of immune activating molecules such as DAMP, ATP, HMGB1 and CRT. These molecules can then activate innate cells capable of eliciting anti-tumor T cell responses and can induce T cell memory. Such T cell responses can be further enhanced by combining with other immune modulators, such as the immune cell engagers described herein. Several of the following examples report the results of studies showing the effectiveness of MMAE in inducing the aforementioned different aspects of the ICD pathway compared to other ADC payloads.

誘導內質網(ER)應激為用於引發免疫原性細胞反應之第一步中之一者，且JNK傳信活化為ER應激之指示符( 參見圖3B)。為評估此指示符，將MIA-PaCa-2胰臟癌細胞用與不同有效負載結合之1 µg/mL ADC處理，如圖7B中所示。在處理24或48小時之後，採集細胞用於西方墨點分析及藉由簡單西方免疫分析(Wes™, Protein Simple)評估上游ER應激標記物pJNK。 Induction of endoplasmic reticulum (ER) stress is one of the first steps for eliciting an immunogenic cellular response, and activation of JNK signaling is an indicator of ER stress ( see Figure 3B). To assess this indicator, MIA-PaCa-2 pancreatic cancer cells were treated with 1 μg/mL ADC combined with different payloads, as shown in Figure 7B. After 24 or 48 hours of treatment, cells were harvested for western blot analysis and assessment of the upstream ER stress marker pJNK by simple western immunoassay (Wes™, Protein Simple).

使用細胞刮刀自培養盤分離經處理細胞。將懸浮細胞以1000 rpm，4℃離心10分鐘。移除上清液且使細胞沈澱物再懸浮於裂解緩衝液(含有蛋白酶及磷酸酶抑制劑)中。在冰上最少10分鐘之後，樣本以13,500 g離心10 min，以沈澱出細胞碎片。將裂解溶液重新定位至各別試管且儲存於-80℃下。使用Bio-Rad DC蛋白質分析套組(目錄號5000112)量化裂解物蛋白質量以允許相等色帶負載。將樣本裂解物及試劑裝載至分析盤中且置放於Wes™中。磷酸化-JNK係使用一級抗體(Cell Signaling Technologies目錄號9251S)鑑別且使用HRP結合之二級抗體(Protein Simple目錄號042-206)及化學發光受質進行免疫探測。所得化學發光信號經偵測、定量且藉由整合式Compass軟體呈現。 7.2 結果 Treated cells were detached from the culture dish using a cell scraper. The cells in suspension were centrifuged at 1000 rpm, 4°C for 10 minutes. The supernatant was removed and the cell pellet was resuspended in lysis buffer (containing protease and phosphatase inhibitors). After a minimum of 10 minutes on ice, samples were centrifuged at 13,500 g for 10 minutes to pellet cellular debris. Lysis solutions were relocated to individual tubes and stored at -80°C. Lysate protein amounts were quantified using the Bio-Rad DC Protein Assay Kit (Cat. No. 5000112) to allow for equal band loading. The sample lysate and reagents are loaded into the assay tray and placed in the Wes™. Phospho-JNK was identified using a primary antibody (Cell Signaling Technologies cat. no. 9251S) and immunoprobed using an HRP-conjugated secondary antibody (Protein Simple cat. no. 042-206) and a chemiluminescent substrate. The resulting chemiluminescent signal is detected, quantified and presented by the integrated Compass software. 7.2 Results

如圖7C至圖7F中所示，在所測試之不同ADC有效負載中，MMAE-ADC (SGD-1006)處理為JNK磷酸化(早期ER應激反應)之最強誘導子之一。一般而言，與用美登素-ADC (圖7C)、喜樹鹼-ADC (圖7D)、蒽環黴素-ADC (圖7E)及卡奇黴素-ADC (圖7F)進行之處理相比，MMAE-ADC處理產生較強pJNK信號。(圖7C至圖7F中之hIgG為與對應ADC具有相同有效負載之非靶向結合物。) 唯一的例外係用含有蒽環黴素mp-EDA-PNU (SGD-8335)之ADC進行之處理，其產生與用MMAE-ADC進行處理的信號相當的pJNK信號(圖7E)。ER應激誘導與微管破壞相關聯，此係由於ER需要完整微管來擴增及收縮以適應細胞之蛋白質轉譯需要。MMAE作為微管蛋白干擾劑誘導此ER應激之能力經例示於所示資料中。 實例 8 ： 對於 MMAE ADC ， 後期 ER 應激標記物之誘導 (CHOP 誘導 ) 一般為優異的 8.1 物質與方法 As shown in Figures 7C-7F, MMAE-ADC (SGD-1006) treatment was one of the strongest inducers of JNK phosphorylation (early ER stress response) among the different ADC payloads tested. In general, treatment with maytansine-ADC (FIG. 7C), camptothecin-ADC (FIG. 7D), anthracycline-ADC (FIG. 7E) and calicheamicin-ADC (FIG. 7F) In contrast, MMAE-ADC treatment produced stronger pJNK signal. (The hIgG in Figures 7C-7F is a non-targeting conjugate with the same payload as the corresponding ADC.) The only exception was treatment with ADC containing anthracycline mp-EDA-PNU (SGD-8335) , which produced a pJNK signal comparable to that processed with the MMAE-ADC (Fig. 7E). ER stress induction is associated with microtubule disruption, as the ER requires intact microtubules to expand and contract to accommodate the protein translation needs of the cell. The ability of MMAE to induce this ER stress as a tubulin interfering agent is exemplified in the data shown. Example 8 : Induction of markers of late ER stress ( CHOP induction ) is generally excellent for MMAE ADCs 8.1 Substances and Methods

CHOP為ER應激反應級聯中之最後一步且其表現量藉由ER應激增加(參見圖3B)。使用經CHOP驅動之螢光素酶報導體(Signosis, Inc.)轉導之MIA-PaCa-2細胞執行ER應激之此下游路徑的評估。包含不同有效負載之若干ADC用於評估。 參見圖7A。 CHOP is the last step in the ER stress response cascade and its expression is increased by ER stress (see Figure 3B). Assessment of this downstream pathway of ER stress was performed using MIA-PaCa-2 cells transduced with a CHOP-driven luciferase reporter (Signosis, Inc.). Several ADCs containing different payloads were used for evaluation. See Figure 7A.

藉由偵測螢光素酶信號(Bright-Glo™螢光素酶分析系統，Promega)量測MIA-PaCa-2細胞中之CHOP誘導。簡言之，將10,000個細胞/孔以75微升/孔接種於96孔黑壁平底透明培養盤中。ADC係以25微升/孔給藥以達成最終IC 50濃度。在36、48及72小時，自培育箱移出培養盤且使其達至室溫。將100 µL Bright-Glo試劑添加至各孔。在讀取之前，搖晃培養盤至少五分鐘。使用Envision CTG 96孔標準協定以讀取培養盤。 8.2 結果 CHOP induction in MIA-PaCa-2 cells was measured by detecting luciferase signal (Bright-Glo™ Luciferase Assay System, Promega). Briefly, 10,000 cells/well were seeded at 75 microliters/well in 96-well black-walled flat-bottom clear culture dishes. ADCs were dosed at 25 microliters/well to achieve final IC50 concentrations. At 36, 48 and 72 hours, the plates were removed from the incubator and allowed to come to room temperature. Add 100 µL Bright-Glo Reagent to each well. Shake the plate for at least five minutes before reading. Plates were read using an Envision CTG 96-well standard protocol. 8.2 Results

如圖8A至圖8D中所示，用含有vc-MMAE (SGD-1006)之ADC進行之處理產生與用含有美登素(SPP-5351)或拉夫坦辛(ravtansine) (SPDB-5352)之ADC進行之處理(圖8A)及用含有mp-EDA-PNU (SGD-8335)或mp-Gluc-DXZ (SGD-8248)之ADC進行之處理(圖8C)的CHOP誘導相當的CHOP誘導。此外，用含有MMAE (SGD-1006)之ADC進行之處理產生比用含有喜樹鹼(圖8B)、AT (SGD-4830) (圖8D)或特絲胺酸(SGD-7455) (圖8D)之ADC進行之處理的CHOP誘導更強的CHOP誘導。相比之下，用含有奧佐黴素(SGD-8677)之ADC進行之處理產生比用含有vc-MMAE(SGD-1006)之ADC進行之處理的CHOP誘導(圖8D)略微更強的CHOP誘導。 實例 9 ： 針對 MMAE ADC ， 免疫刺激 DAMP 之誘導一般為優異的 9.1 物質與方法 As shown in Figures 8A-8D, treatment with ADCs containing vc-MMAE (SGD-1006) resulted in a Treatment with ADC (FIG. 8A) and treatment with ADC containing mp-EDA-PNU (SGD-8335) or mp-Gluc-DXZ (SGD-8248) (FIG. 8C) induced comparable CHOP induction. In addition, treatment with ADCs containing MMAE (SGD-1006) yielded better results than treatment with ADCs containing camptothecin (FIG. 8B), AT (SGD-4830) (FIG. 8D) or teserine (SGD-7455) (FIG. 8D). ) treated with ADCs induced stronger CHOP induction. In contrast, treatment with ADCs containing oxozomycin (SGD-8677) resulted in slightly stronger induction of CHOP than treatment with ADCs containing vc-MMAE (SGD-1006) (FIG. 8D) induce. Example 9 : Induction of immunostimulatory DAMPs is generally excellent for MMAE ADCs 9.1 Substances and Methods

ICD引起釋放免疫活化分子—損傷相關分子模式(DAMP) —諸如ATP、HMGB1及CRT。為量測ICD，如下評估ATP及HMGB1釋放。將MIA-PaCa-2癌細胞用IC 50濃度之具有各種有效負載之ADC處理，以評估活體外ICD標記物誘導。 參見圖7A。 ICDs cause the release of immune-activating molecules—damage-associated molecular patterns (DAMPs)—such as ATP, HMGB1, and CRT. To measure ICD, ATP and HMGB1 release were assessed as follows. MIA-PaCa-2 cancer cells were treated with IC50 concentrations of ADCs with various payloads to assess in vitro ICD marker induction. See Figure 7A.

將200,000個細胞接種於6孔TC培養盤中之每孔中且使其附著至培養盤ON。細胞達到50至60%匯合。將IC 50濃度之ADC添加至各處理孔。在72小時之後，收集500 µL (用於ATP釋放分析)或750 µL (用於HMGB1分析)培養上清液且轉移至經標記1.5 mL埃彭道夫管中。各樣本試管以13,000 rpm離心1分鐘。將50 µL培養基一式三份轉移至96孔透明底培養盤之各孔。將50 µL CellTiter-Glo® (Promega)添加至各孔。搖晃培養盤1至2分鐘。在Envision盤式讀取器上使用CTG96孔標準協定以讀取培養盤。接著，將各試管樣本上清液用於量測HMGB1釋放量，藉由ELISA (IBL)量化該等HMGB1釋放量。HMGB1及ATP釋放量經報導為相對於未經處理樣本之背景值的倍數變化。將所獲取的值轉化成正文檔案且使用Excel及GraphPad Prism導出並分析。 9.2 結果 200,000 cells were seeded in each well of a 6-well TC plate and allowed to attach to the plate ON. Cells reach 50 to 60% confluence. ADCs at IC50 concentrations were added to each treatment well. After 72 hours, 500 µL (for ATP release assay) or 750 µL (for HMGB1 assay) culture supernatant was collected and transferred to labeled 1.5 mL Eppendorf tubes. Each sample tube was centrifuged at 13,000 rpm for 1 minute. Transfer 50 µL of medium in triplicate to each well of a 96-well clear bottom culture plate. 50 µL of CellTiter-Glo® (Promega) was added to each well. Shake the plate for 1 to 2 minutes. Plates were read using the CTG 96-well standard protocol on an Envision plate reader. Next, the supernatant of each test tube sample was used to measure the amount of HMGB1 released, which was quantified by ELISA (IBL). HMGB1 and ATP release are reported as fold changes relative to background values for untreated samples. The acquired values were converted into text files and exported and analyzed using Excel and GraphPad Prism. 9.2 Results

如圖9A至圖9D中所示，用含有vc-MMAE (SGD-1006)之ADC進行之處理引起ATP釋放及HMGB1釋放，其強於用含有類美登素之ADC進行之處理的釋放(圖9A)及用含有喜樹鹼之ADC進行之處理的釋放(圖9B)。此外，用含有MMAE (SGD-1006)之ADC進行之處理引起ATP釋放及HMGB1釋放，其強於用含有特絲胺酸(SGD-7455)或奧瑞他汀AT (SGD-4830)之ADC進行之處理的釋放(圖9D)。 As shown in Figures 9A-9D, treatment with ADCs containing vc-MMAE (SGD-1006) resulted in ATP release and HMGB1 release that were stronger than those with maytansinoid-containing ADCs (Fig. 9A) and release from treatment with ADC containing camptothecin (FIG. 9B). Furthermore, treatment with ADCs containing MMAE (SGD-1006) resulted in ATP release and HMGB1 release that were stronger than those with ADCs containing teserine (SGD-7455) or auristatin AT (SGD-4830). Release of treatment (Fig. 9D).

用含有vc-MMAE (SGD-1006)之ADC進行之處理引起ATP釋放及HMGB1釋放，其與用含有蒽環黴素之ADC (與HMGB1相比，ATP釋放不太穩固)進行之處理的釋放(圖9C)及用含有奧佐黴素(SGD-8677)之ADC進行之處理的釋放(圖9D)相當。 實例 10 ：針對 MMAE ADC ， 先天細胞之活化 ( 細胞介素釋放 ) 一般為優異的 10.1 物質與方法 Treatment with ADCs containing vc-MMAE (SGD-1006) resulted in ATP release and HMGB1 release that was comparable to treatment with anthracycline-containing ADCs (ATP release was less robust compared to HMGB1) ( Figure 9C) was comparable to the release (Figure 9D) from treatment with ADCs containing oxamycin (SGD-8677). Example 10 : Activation of innate cells ( interleukin release ) is generally excellent for MMAE ADCs 10.1 Substances and Methods

DAMP活化可引發抗腫瘤T細胞反應之先天細胞。舉例而言，其可增大單核球上之MHCII表現且釋放先天細胞介素，諸如CXCL-10/IP10。MHCII表現及CXCL-10/IP10評估如下。DAMPs activate innate cells that elicit anti-tumor T cell responses. For example, it can increase MHCII expression on monocytes and release innate interleukins, such as CXCL-10/IP10. MHCII performance and CXCL-10/IP10 assessment are as follows.

洗滌暴露於IC 50濃度之ADC或太平洋紫杉醇24小時(在37℃下，在5%CO 2中)之L540cy癌細胞，且將0.2×10 6個細胞/孔之PBMC以1:10 L540cy:PBMC比率添加至殺死的癌細胞中。用於此實驗之ADC的有效負載描述於圖7A中。將共培養物培育48小時。在第24小時收集細胞培養物上清液，且藉由Luminex評估量測細胞介素，包括先天細胞介素CXCL-10/IP10。 L540cy cancer cells exposed to IC50 concentrations of ADC or paclitaxel for 24 hours (at 37°C in 5% CO2 ) were washed and 0.2 x 106 cells/well of PBMCs were mixed with 1:10 L540cy :PBMCs rate added to the killed cancer cells. The payload of the ADC used for this experiment is depicted in Figure 7A. The co-cultures were incubated for 48 hours. Cell culture supernatants were collected at 24 hours and interferons, including the innate interleukin CXCL-10/IP10, were measured by Luminex assessment.

48小時共培養培育之後，將細胞沈澱物再懸浮於50 μL BD FACs緩衝液中且轉移至96孔圓底微量滴定盤。Fc受體在冰上以100 µg/mL經用人類Fc片段封端，保持30分鐘。在含有100 μg/mL人類純化Fc片段之BD FACs緩衝液中製備以1:100稀釋之包括PE-Cy7抗HLA-DR (MHCII)、PE抗CD14、PE-Dazzle 594抗CD11b、BV605抗CD3及BV421抗CD19的主混合物。將10 µL主混合物添加至含有90 µL再懸浮細胞之各孔中且在冰上培育樣本1小時。接著細胞在預先冷卻之Eppendorf 5810R離心機中以400 xg離心5分鐘。移除上清液且用200 mL BD FACs緩衝液洗滌細胞。執行洗滌兩次且將細胞再懸浮於200 mL FACs緩衝液中。在Attune流式細胞儀上分析樣本。單核球經定義為CD14+CD11b+CD3-CD19-。使用FlowJo分析軟體測定HLA-DR平均螢光。 10.2 結果 After 48 hours of co-culture incubation, the cell pellet was resuspended in 50 μL of BD FACs buffer and transferred to a 96-well round bottom microtiter plate. Fc receptors were capped with human Fc fragments at 100 µg/mL for 30 minutes on ice. Prepared at 1:100 dilution in BD FACs buffer containing 100 μg/mL purified human Fc fragment including PE-Cy7 anti-HLA-DR (MHCII), PE anti-CD14, PE-Dazzle 594 anti-CD11b, BV605 anti-CD3 and Master mix of BV421 anti-CD19. 10 µL of master mix was added to each well containing 90 µL of resuspended cells and samples were incubated on ice for 1 hour. The cells were then centrifuged at 400 xg for 5 minutes in a pre-cooled Eppendorf 5810R centrifuge. The supernatant was removed and cells were washed with 200 mL of BD FACs buffer. Two washes were performed and cells were resuspended in 200 mL of FACs buffer. Samples were analyzed on an Attune flow cytometer. Monocytes are defined as CD14+CD11b+CD3-CD19-. HLA-DR mean fluorescence was determined using FlowJo analysis software. 10.2 Results

如圖10A至圖10D中所示，用含有MMAE (SGD-1006)之ADC進行之處理引起單核球MHC II表現，其與用含有其他有效負載，包括美登素(圖10A)、喜樹鹼(圖10B)、蒽環黴素(圖10C)及卡奇黴素奧佐黴素(SGD-8677)及PBD特絲胺酸(SGD-7455) (圖10D)的ADC進行之處理之表現相當或高於該表現。用含有MMAE (SGD-1006)之ADC進行之處理亦引起先天細胞介素CXCL-10/IP10釋放，其始終高於用含有相同有效負載之ADC進行之處理的釋放(圖10A至圖10D)。 10.3MMAE ADC 之優異 ICD 潛能之概述 As shown in Figures 10A-10D, treatment with ADCs containing MMAE (SGD-1006) resulted in mononucleolar MHC II expression that was comparable to treatment with ADCs containing other payloads, including maytansine (Figure 10A), Camptotheca Performance of treatment with ADCs of base (FIG. 10B), anthracycline (FIG. 10C) and calicheamicin oxazomycin (SGD-8677) and PBD teserine (SGD-7455) (FIG. 10D) equals or exceeds this performance. Treatment with ADCs containing MMAE (SGD-1006) also resulted in release of the innate interleukin CXCL-10/IP10 that was consistently higher than treatment with ADCs containing the same payload (FIGS. 10A-10D). 10.3 Overview of Excellent ICD Potential of MMAE ADCs

如此等實驗中所示，MMAE-ADC可誘導各種ICD標誌及各種免疫原性細胞反應，包括誘導早期ER應激(例如JNK活化)，誘導晚期ER應激(例如CHOP誘導)，誘導免疫活化分子(例如ATP及HMGB1釋放)及活化先天免疫細胞(例如巨噬細胞活化)。所測試之其他ADC有效負載均不始終誘導此等ICD標誌。圖10E提供了具有不同類型之有效負載的ADC之ICD潛能(如藉由上述標誌所量測)的概述且說明微管蛋白干擾劑，特定言之MMAE之整體優越性。 實例 11 ： 基於 Fc 主鏈與 FcγRIIa 、 FcγRIIb 或 FcγRIIIa 之 不同 FcγR 結合 11.1 物質與方法 As shown in experiments such as these, MMAE-ADCs can induce various ICD hallmarks and various immunogenic cellular responses, including induction of early ER stress (eg, JNK activation), induction of late ER stress (eg, CHOP induction), induction of immune activating molecules (eg ATP and HMGB1 release) and activation of innate immune cells (eg macrophage activation). None of the other ADC payloads tested consistently induced these ICD flags. Figure 10E provides an overview of the ICD potential (as measured by the markers above) of ADCs with different types of payloads and illustrates the overall superiority of tubulin interfering agents, specifically MMAE. Example 11 : Binding of different FcγRs based on Fc backbone to FcγRIIa , FcγRIIb or FcγRIIIa 11.1 Materials and methods

使用流式細胞量測術評估抗體SEA-CD40、APX005M、ADC-1013及塞立路單抗(Selicrelumab) (圖11A)之針對經人類FcγRIIa、FcγRIIb或FcγRIIIa轉染之CHO細胞的FcγR結合。將CHO細胞與用於評估結合之增大濃度之抗體及二級抗體一起培育，如藉由流式細胞量測術監測。FcyR binding of antibodies SEA-CD40, APX005M, ADC-1013 and Selicrelumab (FIG. 11A) to CHO cells transfected with human FcyRIIa, FcyRIIb or FcyRIIIa was assessed using flow cytometry. CHO cells are incubated with increasing concentrations of antibody and secondary antibody for assessing binding, as monitored by flow cytometry.

對於各細胞株，將5千萬個細胞在50 mL PBS中洗滌一次。細胞經再次計數且以220萬個細胞/毫升再懸浮於BD染色緩衝液中。將細胞以0.1毫升細胞/孔接種於96孔圓底盤中。For each cell line, 50 million cells were washed once in 50 mL of PBS. Cells were counted again and resuspended in BD staining buffer at 2.2 million cells/ml. Cells were seeded in 96-well round dishes at 0.1 ml cells/well.

將抗體溶液稀釋形成以下最終濃度：3 mg/mL、1 mg/mL、0.3 mg/mL、0.1 mg/mL、0.03 mg/mL、0.01 mg/mL、0.003 mg/mL、0.001 mg/mL、0.0003 mg/mL。各抗體溶液以10×稀釋(亦即將11 µL各抗體溶液添加至89 µL培養基)，以產生以下濃度：300、100、30、10、3、1、0.03、0.01、0.003、0.001及0.0003 μg/ml。自細胞移除培養基且用培養基洗滌細胞。將100 µL抗體溶液添加至各孔。在豎直方向上以減小的濃度添加抗體溶液。在4℃下培育1小時之後，將培養盤離心且各孔細胞用200 µL BD染色緩衝液洗滌兩次。藉由使培養盤渦旋來再懸浮所沈澱之細胞。Dilute the antibody solution to the following final concentrations: 3 mg/mL, 1 mg/mL, 0.3 mg/mL, 0.1 mg/mL, 0.03 mg/mL, 0.01 mg/mL, 0.003 mg/mL, 0.001 mg/mL, 0.0003 mg/mL. Each antibody solution was diluted 10× (i.e., 11 µL of each antibody solution was added to 89 µL of medium) to yield the following concentrations: 300, 100, 30, 10, 3, 1, 0.03, 0.01, 0.003, 0.001, and 0.0003 μg/ ml. The medium was removed from the cells and the cells were washed with the medium. Add 100 µL of antibody solution to each well. Antibody solutions are added in decreasing concentrations in a vertical direction. After 1 hour incubation at 4°C, the plates were centrifuged and the cells in each well were washed twice with 200 µL of BD staining buffer. The pelleted cells were resuspended by vortexing the culture plate.

接著，在BD染色緩衝液中製備PE結合之抗人類IgG Fc抗體(1/50稀釋之1 mg/ml濃度=33 µg/mL飽和濃度)。在暗冰箱中培育溶液30 min。在培育之後，將培養盤離心，移除上清液且細胞用每孔200 µL BD染色緩衝液洗滌兩次。將細胞再懸浮於PBS + 1%多聚甲醛中且保持在4℃下直至其藉由流式細胞量測術分析。使用Attune流式細胞儀分析樣本。使用Graphpad Prism圖示出螢光強度之幾何平均值(GEO平均螢光)之資料點。 11.2 結果 Next, PE-conjugated anti-human IgG Fc antibody was prepared in BD staining buffer (1 mg/ml concentration at 1/50 dilution = 33 µg/mL saturation concentration). Incubate the solution in a dark refrigerator for 30 min. After incubation, plates were centrifuged, supernatant removed and cells washed twice with 200 µL per well of BD staining buffer. Cells were resuspended in PBS + 1% paraformaldehyde and kept at 4°C until they were analyzed by flow cytometry. Samples were analyzed using an Attune flow cytometer. Data points for the geometric mean of fluorescence intensity (GEO mean fluorescence) were plotted using Graphpad Prism. 11.2 Results

APX005 S267E展現對於FcγRIIa及FcγIIb之最高親和力(圖11B至圖11D)。SEA-CD40對FcγRIIIa具有最高親和力且對FcγRIIb具有最低親和力(圖11B至圖11D)。資料展現不同Fc主鏈影響對與不同FcγR結合之潛能。SEA-CD40未經岩藻糖基化主鏈展示與研發中之其他CD40抗體相比，不同的結合在於其結合活化而非抑制性FcγR (圖11B至圖11D)。 實例 12 ： MIA-PaCa-2 細胞中之誘導細胞死亡 12.1 物質與方法 APX005 S267E exhibited the highest affinity for FcyRIIa and FcyIIb (FIGS. 11B-11D). SEA-CD40 had the highest affinity for FcyRIIIa and the lowest affinity for FcyRIIb (FIG. 11B-11D). The data demonstrate the effect of different Fc backbones on the potential for binding to different FcγRs. SEA-CD40 unfucosylated backbone displays binding differently compared to other CD40 antibodies in development in that it binds activating rather than inhibitory FcγRs (FIGS. 11B-11D). Example 12 : Induction of cell death in MIA-PaCa-2 cells 12.1 Substances and methods

藉由EC50濃度之非ICD誘導劑阿布拉生(Abraxane) (其類似於上述實例3中之太平洋紫杉醇起作用)或2種ICD誘導劑奧沙利鉑或vc-MMAE誘導MIA-PaCa-2胰臟腫瘤細胞之細胞死亡。將細胞與各藥劑一起培育18小時。接著將腫瘤細胞添加至人類PBMC加具有不同Fc主鏈之各種CD40導引之促效劑(1 µg/ml) (圖11A)且48小時後評估免疫活化。 12.2 結果 MIA-PaCa-2 pancreas was induced by EC50 concentrations of the non-ICD inducer Abraxane (which acts similarly to paclitaxel in Example 3 above) or the 2 ICD inducers oxaliplatin or vc-MMAE. Cell death of dirty tumor cells. Cells were incubated with each agent for 18 hours. Tumor cells were then added to human PBMC plus various CD40-directed agonists (1 μg/ml) with different Fc backbones (FIG. 11A) and immune activation assessed 48 hours later. 12.2 Results

SEA-CD40與vcMMAE ADC之組合藉由誘導優異釋放免疫活化細胞介素(CXCL10及IFNγ；圖12A至圖12B)至少部分地殺死腫瘤細胞，而具有不同Fc主鏈之其他CD40促效劑擴增免疫抑制細胞介素(IL-10及MDC；圖12C至圖12D)。此實例說明藉由增強與FcγRIIIa之結合的Fc主鏈觀測到改善的免疫反應。 實例 13 ：針對 細胞凋亡黑素瘤細胞之不同免疫活化隨 Fc 主鏈而變化 13.1 物質與方法 The combination of SEA-CD40 and vcMMAE ADC at least partially kills tumor cells by inducing superior release of immune activating interleukins (CXCL10 and IFNγ; Figures 12A-12B), while other CD40 agonists with different Fc backbones expand Increased immunosuppressive interleukins (IL-10 and MDC; Figures 12C-12D). This example illustrates that an improved immune response is observed with an Fc backbone that enhances binding to FcyRIIIa. Example 13 : Different immune activation against apoptotic melanoma cells as a function of Fc backbone 13.1 Materials and methods

兩種黑素瘤細胞株(SK-MEL 12及SK-MEL 28)用EC50濃度之阿布拉生，草酸鉑(oxaloplatin)或vc-MMAE處理18小時。將人類PBMC加1 µg/ml具有不同Fc主鏈之CD40導引之促效劑(圖11A)添加至經處理之黑素瘤/腫瘤細胞。48小時後評估免疫活化。 13.2 結果 Two melanoma cell lines (SK-MEL 12 and SK-MEL 28) were treated with EC50 concentrations of abrasin, oxaloplatin or vc-MMAE for 18 hours. Human PBMC plus 1 μg/ml of CD40-directed agonists with different Fc backbones (FIG. 11A) were added to treated melanoma/tumor cells. Immune activation was assessed after 48 hours. 13.2 Results

SEA-CD40與vc-MMAE ADC之組合誘導釋放免疫活化細胞介素(CXCL10；圖13A至圖13B)，而其他CD40促效劑擴增免疫抑制細胞介素(IL-10；圖13C至圖13D)。 實例 14 ：針對 多種細胞凋亡腫瘤細胞類型之不同免疫活化隨 Fc 主鏈而變化 14.1 物質與方法 14.1.1 細胞 The combination of SEA-CD40 and vc-MMAE ADC induces the release of immune-activating interleukins (CXCL10; Figures 13A-13B), while other CD40 agonists amplify immune-suppressing interleukins (IL-10; Figures 13C-13D ) ). Example 14 : Different immune activation against various apoptotic tumor cell types as a function of Fc backbone 14.1 Materials and methods 14.1.1 Cells

使用EC50濃度之ICD誘導劑奧沙利鉑或vc-MMAE及培育18小時在來自黑素瘤、肺、乳房及胰臟之腫瘤細胞中誘導細胞死亡。將經處理腫瘤細胞添加至人類PBMC及具有不同Fc主鏈之各種CD40導引之促效劑中(圖11A)。48小時後評估免疫活化。如上文實例5中所描述處理PBMC及腫瘤細胞株。代替MIA-PaCa-2癌細胞(其用於實例5中)，使用以下細胞株：黑素瘤細胞株SK-MEL 12及SK-MEL 28、肺癌細胞株A549、乳癌細胞株MDA-MB-468及胰臟癌細胞株MIA-PaCa-2。一式三份地處理細胞。 14.1.2 細胞介素產生 Cell death was induced in tumor cells from melanoma, lung, breast and pancreas using EC50 concentrations of the ICD inducers oxaliplatin or vc-MMAE and incubation for 18 hours. Treated tumor cells were added to human PBMCs and various CD40-directed agonists with different Fc backbones (FIG. 11A). Immune activation was assessed after 48 hours. PBMC and tumor cell lines were processed as described in Example 5 above. Instead of MIA-PaCa-2 cancer cells (which were used in Example 5), the following cell lines were used: melanoma cell lines SK-MEL 12 and SK-MEL 28, lung cancer cell line A549, breast cancer cell line MDA-MB-468 and pancreatic cancer cell line MIA-PaCa-2. Cells were processed in triplicate. 14.1.2 Cytokinin production

如上文實例5中所描述評估細胞介素產生。 14.2 結果 Interferon production was assessed as described in Example 5 above. 14.2 Results

SEA-CD40與vc-MMAE ADC之組合驅動優異釋放免疫活化細胞介素(CXCL10及IFNγ；圖14A及14C)，而其他CD40促效劑擴增免疫抑制細胞介素(IL-10；圖14B)。如同實例12及13，此實例展現與其他Fc主鏈相比，藉由增強與FcγRIIIa之結合的Fc主鏈觀測到之改善的免疫反應。 實例 15 ： 未經岩藻糖基化 SEA-CD40 抗體與基於奧瑞他汀之 ADC 的組合之協同效果 15.1 物質與方法 The combination of SEA-CD40 and vc-MMAE ADC drives superior release of immune-activating interleukins (CXCL10 and IFNγ; Figures 14A and 14C), while other CD40 agonists amplify immune-suppressive interleukins (IL-10; Figure 14B) . As with Examples 12 and 13, this example demonstrates the improved immune response observed with an Fc backbone that enhances binding to FcyRIIIa compared to other Fc backbones. Example 15 : Synergistic Effects of Combination of Unfucosylated SEA-CD40 Antibody and Auristatin-Based ADC 15.1 Materials and Methods

人類CD40轉殖基因小鼠經植入有經工程改造以表現抗原Thy 1.1之A20細胞。在第0天，將腫瘤細胞皮下植入側腹。當達至100 mm 3平均腫瘤大小(藉由使用公式：體積(mm 3)=0.5*長度*寬度 2量測，其中長度為較長尺寸)時，將小鼠隨機分為5隻小鼠/組之處理組。接著動物用指定的腹膜內處理進行處理；每三天給予各處理一次，總計三次處理。將原液濃度之抗體稀釋至適當濃度且以100 µl體積注射至動物體內。最終劑量針對SEA-CD40為1 mg/kg且針對Thy1.1之vc-MMAE ADC為1 mg/kg。在整個研究期間量測腫瘤長度、腫瘤寬度及小鼠重量，且使用上式計算腫瘤體積。追蹤動物直至量測到腫瘤體積達至約1,000 mm 3，此時使動物安樂死。 15.2 結果 Human CD40 transgenic mice were implanted with A20 cells engineered to express the antigen Thy 1.1. On day 0, tumor cells were implanted subcutaneously into the flank. Mice were randomly divided into 5 mice/ The treatment group of the group. Animals were then treated with the indicated intraperitoneal treatments; each treatment was given every three days for a total of three treatments. The stock concentration of antibody was diluted to the appropriate concentration and injected into animals in a volume of 100 µl. Final doses were 1 mg/kg for SEA-CD40 and 1 mg/kg for vc-MMAE ADC of Thyl.1. Tumor length, tumor width, and mouse weight were measured throughout the study, and tumor volume was calculated using the above formula. Animals were followed until tumor volume was measured to reach approximately 1,000 mm3 , at which point the animals were euthanized. 15.2 Results

如圖15中所示，用次治療劑量之SEA-CD40處理A20腫瘤模型使得腫瘤生長減少且腫瘤生長延遲。類似地，含有vc-MMAE之抗體-藥物結合物展示輕度腫瘤生長延遲。然而，當兩種藥劑經串聯投與至動物時，觀測到治癒性抗腫瘤反應。此資料展現將增強型SEA-CD40抗體與經由ADC遞送之免疫原性細胞死亡誘導化學療法兩者組合之協同益處，包括達成治癒性反應之可能性。 實例 16 ： 用靶向腫瘤相關抗原的基於奧瑞他汀之 ADC 及具有不同 Fc 主鏈之 TIGIT 抗體處理的各種腫瘤細胞株之不同活性 16.1 物質與方法 As shown in Figure 15, treatment of the A20 tumor model with sub-therapeutic doses of SEA-CD40 resulted in decreased tumor growth and delayed tumor growth. Similarly, antibody-drug conjugates containing vc-MMAE exhibited mild tumor growth delay. However, when the two agents were administered to animals in tandem, a curative antitumor response was observed. This data demonstrates the synergistic benefits of combining enhanced SEA-CD40 antibodies with immunogenic cell death-inducing chemotherapy delivered via ADCs, including the potential to achieve a curative response. Example 16 : Different Activities of Various Tumor Cell Lines Treated with Auristatin -Based ADCs Targeting Tumor - Associated Antigens and TIGIT Antibodies with Different Fc Backbone 16.1 Materials and Methods

五種不同人類癌症細胞株：SK-MEL 28 (黑素瘤)、MDA-MB-468 (乳房)、CORL23 (肺)、A549 (肺)及HT-26 (大腸)用於此實例。各細胞株用1 μg/ml藥物抗體比(DAR)為4之靶向腫瘤之抗體-vcMMAE ADC處理18小時。在37℃下培育之後，洗滌腫瘤細胞且添加人類PBMC以及如圖16中所指示之1 μg/ml抗TIGIT抗體處理。所使用之抗TIGIT抗體具有不同程度的主鏈效應功能，其中LALA TIGIT抗體無FcγR結合且SEA-TIGIT抗體具有增強的FcγR結合(增強與活化FcγIIIaR之結合且降低與抑制性FcγRIIbR之結合)。下表D展示不同TGT抗體之相對活性。將免疫細胞、抗TIGIT抗體處理及死亡/正在死亡腫瘤細胞之此等培養物培育額外48小時。採集用於各條件之上清液且使用ELISA根據製造商之說明書評估細胞介素誘導。 表 D ： TGT 抗體    CD155 T reg 消耗 CD8 T 細胞反應 APC 活化 抗TIGIT LALA ++ - - - 抗TIGIT IgG1 ++ ++ ++ + SEA-TGT ++ ++++ +++ +++ 16.2 結果 Five different human cancer cell lines: SK-MEL 28 (melanoma), MDA-MB-468 (breast), CORL23 (lung), A549 (lung) and HT-26 (large intestine) were used for this example. Each cell line was treated with 1 μg/ml drug-to-antibody ratio (DAR) of 4, a tumor-targeting antibody-vcMMAE ADC for 18 hours. After incubation at 37°C, tumor cells were washed and human PBMCs were added and treated with 1 μg/ml anti-TIGIT antibody as indicated in FIG. 16 . The anti-TIGIT antibodies used had varying degrees of backbone effector function, with the LALA TIGIT antibody having no FcyR binding and the SEA-TIGIT antibody having enhanced FcyR binding (enhanced binding to activating FcyIIIaR and decreased binding to inhibitory FcyRIIbR). Table D below shows the relative activities of different TGT antibodies. These cultures of immune cells, anti-TIGIT antibody treatment and dead/dying tumor cells were incubated for an additional 48 hours. Supernatants for each condition were collected and assessed for cytokine induction using ELISA according to the manufacturer's instructions. Table D : TGT Antibodies CD155 T reg consumption CD8 T cell response APC activation Anti-TIGIT LALA ++ - - - anti-TIGIT IgG1 ++ ++ ++ + SEA-TGT ++ ++++ +++ +++ 16.2 Results

如圖16中所示，在不包含任何其他免疫調節處理之PBMC培育(圖16中之「未經處理」)下的死亡/正在死亡腫瘤細胞具有一些免疫細胞刺激，如先天I型干擾素相關之細胞介素IP10之產生所見。IP10活化程度視腫瘤細胞類型而定，此係由於SK-MEL 28、MDA-MB-468及CORL23比A549或HT-26細胞誘導更強PBMC活化。然而，不管腫瘤細胞，隨後將增強效應功能之抗TIGIT mAb SEA-TGT添加至共培養物引起整個板之免疫細胞活化之進一步增強。對於僅與死亡細胞共培育之細胞株不足以驅動顯著的活化，包括未經岩藻糖基化TIGIT mAb SEA-TGT展示最顯著的增加，其表明其較強活化能力。在此等細胞中，IgG1主鏈TIGIT抗體能夠驅動在與增強之mAb相比時減弱的免疫細胞活化。不具有FcγR結合能力之抗TIGIT之LALA版本在驅動任何免疫活化時無活性。 實例 17 ： 用靶向腫瘤相關抗原的基於奧瑞他汀之 ADC 及具有不同 Fc 主鏈之 TIGIT 抗體處理的各種腫瘤細胞株之不同活性 17.1 物質與方法 As shown in Figure 16, dead/dying tumor cells incubated with PBMCs without any other immunomodulatory treatments ("untreated" in Figure 16) had some immune cell stimulation, such as innate type I interferon-related The production of interleukin IP10 was seen. The extent of IP10 activation was tumor cell type dependent, as SK-MEL 28, MDA-MB-468 and CORL23 induced stronger PBMC activation than A549 or HT-26 cells. However, subsequent addition of the effector-enhancing anti-TIGIT mAb SEA-TGT to the co-culture resulted in a further enhancement of immune cell activation across the plate, regardless of tumor cells. Co-culture with dead cells alone was not sufficient to drive significant activation, including the unfucosylated TIGIT mAb SEA-TGT showed the most significant increase, indicating its stronger activation capacity. In these cells, the IgGl backbone TIGIT antibody was able to drive attenuated immune cell activation when compared to the boosted mAb. The LALA version of anti-TIGIT, which has no Fc[gamma]R binding ability, is inactive in driving any immune activation. Example 17 : Different Activities of Various Tumor Cell Lines Treated with Auristatin -Based ADCs Targeting Tumor - Associated Antigens and TIGIT Antibodies with Different Fc Backbone 17.1 Materials and Methods

六種不同人類癌症細胞株HT-26 (大腸)、A549 (肺)、CORL23 (肺)、MDA-MB-468 (乳房)、SK-MEL 28 (黑素瘤)及Mia-PaCa-2 (胰臟)用於此實例。各細胞株用1 μg/ml藥物抗體比(DAR)為4之靶向腫瘤之抗體-vcMMAE ADC處理18小時。在37℃下培育之後，洗滌腫瘤細胞且添加人類PBMC以及如圖16中所指示之1 μg/ml抗TIGIT抗體處理。將免疫細胞、抗TIGIT抗體處理及死亡/正在死亡腫瘤細胞之此等培養物培育額外48小時。採集用於各條件之上清液且使用ELISA根據製造商之說明書評估細胞介素誘導。 17.2 結果 Six different human cancer cell lines HT-26 (large intestine), A549 (lung), CORL23 (lung), MDA-MB-468 (breast), SK-MEL 28 (melanoma) and Mia-PaCa-2 (pancreas) dirty) for this instance. Each cell line was treated with 1 μg/ml drug-to-antibody ratio (DAR) of 4, the tumor-targeting antibody-vcMMAE ADC for 18 hours. After incubation at 37°C, tumor cells were washed and human PBMCs were added and treated with 1 μg/ml anti-TIGIT antibody as indicated in FIG. 16 . These cultures of immune cells, anti-TIGIT antibody treatment and dead/dying tumor cells were incubated for an additional 48 hours. Supernatants for each condition were collected and assessed for cytokine induction using ELISA according to the manufacturer's instructions. 17.2 Results

如圖17中所示，在不包含任何其他免疫調節處理之PBMC培育(圖16中之「未經處理」)下的死亡/正在死亡腫瘤細胞具有一些免疫細胞刺激，如自適應細胞介素IFNγ之產生所見。IFNγ活化程度視腫瘤細胞類型而定，此係由於SK-MEL 28、MDA-MB-468及CORL23比A549或HT-26細胞誘導更強PBMC活化。然而，不管腫瘤細胞，隨後將增強效應功能之未經岩藻糖基化SEA-TGT mAb添加至共培養物中引起整個板之免疫細胞活化之進一步增強。對於僅與死亡細胞共培育之細胞株不足以驅動顯著的活化，包括未經岩藻糖基化TIGIT mAb SEA-TGT展示最顯著的增加，其表明其較強活化能力。在此等細胞中，IgG1主鏈TIGIT抗體能夠驅動一定免疫細胞活化，但其與增強之mAb相比時極大地減弱。不具有FcγR結合能力之抗TIGIT之LALA版本在驅動任何免疫活化時無活性。 實例 18 ： 未經岩藻糖基化 TIGIT 抗體與基於奧瑞他汀之 ADC 的組合之協同效果 18.1 物質與方法 18.1.1TIGIT抗體及基於奧瑞他汀之ADC的活體外評估 As shown in Figure 17, dead/dying tumor cells incubated with PBMCs without any other immunomodulatory treatments ("untreated" in Figure 16) had some immune cell stimulation, such as the adaptive interferon IFNy produced what is seen. The extent of IFNγ activation was dependent on tumor cell type, as SK-MEL 28, MDA-MB-468 and CORL23 induced stronger PBMC activation than A549 or HT-26 cells. However, subsequent addition of the non-fucosylated SEA-TGT mAb, which enhances effector function, to the co-culture resulted in a further enhancement of immune cell activation across the plate, regardless of tumor cells. Co-incubation with dead cells alone was not sufficient to drive significant activation, including unfucosylated TIGIT mAb SEA-TGT showed the most significant increase, indicating its stronger activation capacity. In these cells, the IgG1 backbone TIGIT antibody was able to drive some immune cell activation, but it was greatly attenuated when compared to the boosted mAb. The LALA version of anti-TIGIT, which has no Fc[gamma]R binding ability, is inactive in driving any immune activation. Example 18 : Synergistic Effects of Combination of Unfucosylated TIGIT Antibodies and Auristatin- Based ADCs 18.1 Materials and Methods 18.1.1 In Vitro Evaluation of TIGIT Antibodies and Auristatin-Based ADCs

用EC 50濃度之ICD誘導劑vc-MMAE與腫瘤細胞靶向抗體之結合誘導A549非小細胞肺癌瘤細胞的細胞死亡。將細胞與藥劑一起培育18小時且接著與各種濃度(1、0.1、0.01 μg/ml)之具有不同Fc主鏈之抗TIGIT抗體，包括抗TIGIT LALA、SEA-TGT及抗體31C6 H4/L1 (其為IgG1抗體) (US 2018/0066055 A1)共同添加至人類PBMC中。接著，藉由在共培養48小時之後量測細胞介素(IP10)含量評估免疫活化。 18.1.2TIGIT抗體及基於奧瑞他汀之ADC的活體內評估 Cell death of A549 non-small cell lung cancer cells was induced with the binding of the ICD inducer vc-MMAE at EC50 concentrations to tumor cell targeting antibodies. Cells were incubated with agents for 18 hours and then with various concentrations (1, 0.1, 0.01 μg/ml) of anti-TIGIT antibodies with different Fc backbones, including anti-TIGIT LALA, SEA-TGT, and antibody 31C6 H4/L1 (which IgG1 antibody) (US 2018/0066055 A1) was co-added to human PBMC. Next, immune activation was assessed by measuring interleukin (IP10) content after 48 hours of co-culture. 18.1.2 In vivo evaluation of TIGIT antibodies and auristatin-based ADCs

Balb/c小鼠在第0天經皮下在側腹植入表現腫瘤抗原Thy1.1之CT26同基因型腫瘤細胞株。當達至100 mm 3平均腫瘤大小(藉由使用公式：體積(mm 3)=0.5*長度*寬度 2量測，其中長度為較長尺寸)時，將小鼠隨機分為5隻小鼠/組之處理組。接著動物用指定的腹膜內處理進行處理；每三天給予各處理一次，總計三次處理。將原液濃度之抗體稀釋至適當濃度且以100 µl體積注射至動物體內。最終劑量針對SEA-TGT mIgG2a為0.1 mg/kg且針對靶向腫瘤之vc-MMAE Thy1.1 ADC為5 mg/kg。所使用的兩種抗體均在mIgG2a主鏈上且SEA-TGT mIgG2a未經岩藻糖基化。在整個研究期間量測腫瘤長度、腫瘤寬度及小鼠重量，且使用上式計算腫瘤體積。追蹤動物直至量測到腫瘤體積達至約1,000 mm 3，此時使動物安樂死。 Balb/c mice were implanted subcutaneously in the flank on day 0 with a CT26 isogenic tumor cell line expressing the tumor antigen Thyl.1. Mice were randomly divided into 5 mice/ The treatment group of the group. Animals were then treated with the indicated intraperitoneal treatments; each treatment was given every three days for a total of three treatments. The stock concentration of antibody was diluted to the appropriate concentration and injected into animals in a volume of 100 µl. Final doses were 0.1 mg/kg for SEA-TGT mIgG2a and 5 mg/kg for tumor-targeted vc-MMAE Thyl.1 ADC. Both antibodies used were on the mIgG2a backbone and SEA-TGT mIgG2a was not fucosylated. Tumor length, tumor width, and mouse weight were measured throughout the study, and tumor volume was calculated using the above formula. Animals were followed until tumor volume was measured to reach approximately 1,000 mm3 , at which point the animals were euthanized.

Balb/c小鼠在第0天經皮下在側腹植入表現腫瘤抗原EphA2之Renca同基因型腫瘤細胞株。當達至100 mm 3平均腫瘤大小(藉由使用公式：體積(mm 3)=0.5*長度*寬度 2量測，其中長度為較長尺寸)時，將小鼠隨機分為5隻小鼠/組之處理組。接著動物用指定的腹膜內處理進行處理；每三天給予各處理一次，總計三次處理。將原液濃度之抗體稀釋至適當濃度且以100 µl體積注射至動物體內。最終劑量針對SEA-TGT為0.1 mg/kg且針對靶向腫瘤之vc-MMAE EphA2 ADC為1 mg/kg。所使用的兩種抗體均在mIgG2a主鏈上。在整個研究期間量測腫瘤長度、腫瘤寬度及小鼠重量，且使用上式計算腫瘤體積。追蹤動物直至量測到腫瘤體積達至約1,000 mm 3，此時使動物安樂死。 18.2 結果 Balb/c mice were implanted subcutaneously on day 0 with Renca isogenic tumor cell lines expressing the tumor antigen EphA2 in the flanks. Mice were randomly divided into 5 mice/ The treatment group of the group. Animals were then treated with the indicated intraperitoneal treatments; each treatment was given every three days for a total of three treatments. The stock concentration of antibody was diluted to the appropriate concentration and injected into animals in a volume of 100 µl. Final doses were 0.1 mg/kg for SEA-TGT and 1 mg/kg for tumor-targeted vc-MMAE EphA2 ADC. Both antibodies used were on the mIgG2a backbone. Tumor length, tumor width, and mouse weight were measured throughout the study, and tumor volume was calculated using the above formula. Animals were followed until tumor volume was measured to reach approximately 1,000 mm3 , at which point the animals were euthanized. 18.2 Results

如圖18A中所示，將ADC殺死之腫瘤細胞與免疫細胞群一起培育引起一定免疫細胞活化，此係因為經由MMAE誘導之免疫原性細胞死亡，如藉由細胞介素IP10之誘導所量測(參見X軸上標記有「0」之條柱)。此外，將增大濃度之SEA-TGT添加至共培養物中引起此細胞介素誘導之顯著增強。此增強在效應子剔除式抗TIGIT抗體(LALA)或標準抗TIGIT IgG1抗體(31C6 H4/L1)情況下不可見，從而展示出SEA-TGT之未經岩藻糖基化主鏈與誘導免疫原性細胞死亡之MMAE協同驅動以提供優異免疫細胞活化。As shown in Figure 18A, incubation of ADC-killed tumor cells with immune cell populations resulted in some immune cell activation due to immunogenic cell death induced by MMAE, as measured by induction of interferon IP10 measurement (see the bar marked "0" on the x-axis). Furthermore, the addition of increasing concentrations of SEA-TGT to the co-cultures resulted in a significant enhancement of this cytokine induction. This enhancement was not seen with the effector knockout anti-TIGIT antibody (LALA) or the standard anti-TIGIT IgG1 antibody (31C6 H4/L1), demonstrating the unfucosylated backbone of SEA-TGT and the inducing immunogen MMAE synergistic drive of sexual cell death to provide superior immune cell activation.

如圖18B及圖18C中所示，藉由次治療劑量之0.1 mg/kg SEA-TGT分別處理CT26腫瘤模型及Renca腫瘤模型使得腫瘤生長減少且腫瘤生長延遲。vc-MMAE (維多汀) ADC獨自展示輕度腫瘤生長延遲。當將兩種藥劑以串聯方式投與至動物時，觀測到腫瘤生長之顯著減小以及40%動物之治癒性反應。此等資料展現將具有增強效應功能之SEA-TGT mIgG2a抗體(重新格式化為對應於未經岩藻糖基化人類IgG1主鏈之未經岩藻糖基化小鼠IgG2a的SEA-TGT抗體)與誘導免疫原性細胞死亡之ADC組合之協同性益處。As shown in Figure 18B and Figure 18C, treatment of the CT26 tumor model and the Renca tumor model with a sub-therapeutic dose of 0.1 mg/kg SEA-TGT, respectively, resulted in decreased tumor growth and delayed tumor growth. The vc-MMAE (vedotin) ADC alone exhibited mild tumor growth delay. When the two agents were administered to animals in tandem, a significant reduction in tumor growth and a curative response in 40% of the animals were observed. These data demonstrate that SEA-TGT mIgG2a antibody with enhanced effector function (reformatted as SEA-TGT antibody corresponding to unfucosylated mouse IgG2a without fucosylated human IgG1 backbone) Synergistic benefits in combination with ADCs that induce immunogenic cell death.

在兩種不同腫瘤模型中藉由靶向兩種不同腫瘤細胞抗原之ADC得到此類觀察結果之事實表明此組合之抗腫瘤活性可廣泛適用於不同腫瘤類型。 實例 19 ： 未經岩藻糖基化 TIGIT 抗體與另一基於奧瑞他汀之 ADC 的組合之協同效果 19.1 物質與方法 The fact that such observations were obtained with ADCs targeting two different tumor cell antigens in two different tumor models suggests that the antitumor activity of this combination is broadly applicable to different tumor types. Example 19 : Synergistic Effect of Combination of Unfucosylated TIGIT Antibody and Another Auristatin-Based ADC 19.1 Materials and Methods

經工程改造以表現鼠類B7H4之Renca細胞經皮下植入於Balb/c小鼠體內。使腫瘤生長達至100 mm 3，此時用次治療劑量之SEA-TGT及SGN-B7H4 MMAE ADC (B7H4V)或次治療劑量之SEA-TGT及治療性劑量之奧沙利鉑處理小鼠。在同一天給予各化合物，且相隔7天處理小鼠，總計3個劑量。 19.2 結果 Renca cells engineered to express murine B7H4 were implanted subcutaneously in Balb/c mice. Tumors were allowed to grow to 100 mm3 , at which point mice were treated with sub-therapeutic doses of SEA-TGT and SGN-B7H4 MMAE ADC (B7H4V) or sub-therapeutic doses of SEA-TGT and therapeutic doses of oxaliplatin. Compounds were administered on the same day and mice were treated 7 days apart for a total of 3 doses. 19.2 Results

如圖19中所示，當次治療劑量之SEA-TGT與次治療劑量之B7H4V組合時，SEA-TGT組合活性經擴展以增大抗腫瘤活性。SEA-TGT (次治療劑量)與B7H4V (次治療劑量)之組合活性與SEA-TGT(次治療劑量)及已知ICD誘導劑之奧沙利鉑(治療劑量)之組合活性類似。然而，奧沙利鉑與全身性過敏反應及腎毒性之警告相關，且當以治療劑量給予時，其自身並不起活性且通常與多種其他化學療法組合使用。As shown in Figure 19, when a sub-therapeutic dose of SEA-TGT was combined with a sub-therapeutic dose of B7H4V, the SEA-TGT combination activity was extended to increase anti-tumor activity. The combined activity of SEA-TGT (subtherapeutic dose) and B7H4V (subtherapeutic dose) was similar to the combined activity of SEA-TGT (subtherapeutic dose) and oxaliplatin (therapeutic dose), a known ICD inducer. However, oxaliplatin is associated with warnings of anaphylaxis and nephrotoxicity, and when given in therapeutic doses, is not active on its own and is often used in combination with various other chemotherapies.

亦如圖19中所示，SEA-TGT在與B7H4V組合時之治癒性效果顯著增大。儘管不意欲受理論束縛，但認為此效果係歸因於ICD誘導及用此類組合誘導之長期記憶T細胞反應(亦見下方實例23)。As also shown in Figure 19, the curative effect of SEA-TGT was significantly increased when combined with B7H4V. While not intending to be bound by theory, it is believed that this effect is due to ICD induction and long-term memory T cell responses induced with such combinations (see also Example 23 below).

綜上所述，此實驗之結果符合本文所描述之其他結果，其展示出針對免疫細胞接合子之未經岩藻糖基化抗體(在此實驗中，未經岩藻糖基化抗TIGIT抗體，諸如SEA-TGT)與誘導免疫細胞死亡之藥劑很好地組合，該藥劑在此特定實例中為MMAE ADC (亦即B7H4V)及奧沙利鉑兩者。然而，與MMAE ADC之組合為較佳的，此係因為與奧沙利鉑相關聯之毒性且因為與治療劑量之奧沙利鉑相比，在次治療劑量之ADC下觀測到相當的治療效果。 實例 20 ： 未經岩藻糖基化 SEA-CD70 抗體與基於奧瑞他汀之 ADC 的組合之協同效果 Taken together, the results of this experiment are in line with other results described herein showing non-fucosylated antibodies against immune cell zygotes (in this experiment, non-fucosylated anti-TIGIT antibodies , such as SEA-TGT) is well combined with an agent that induces immune cell death, which in this particular example is both MMAE ADC (ie, B7H4V) and oxaliplatin. However, the combination with MMAE ADC is preferred because of the toxicity associated with oxaliplatin and because comparable therapeutic effects are observed at sub-therapeutic doses of ADC compared to therapeutic doses of oxaliplatin . Example 20 : Synergistic effect of combination of non-fucosylated SEA-CD70 antibody and auristatin-based ADC

SEA-CD70 (SEA-h1F6)為靶向CD70抗原之未經岩藻糖基化抗體。CD70分子為腫瘤壞死因子(TNF)配位體超家族(TNFSF)中之成員且其結合於相關受體CD27 (TNFRSF7)。兩個分子之間的相互作用活化來自兩種受體之細胞內信號。在正常條件下，CD70表現為短暫的且限於活化之T及B細胞、成熟樹突狀細胞及自然殺手(NK)細胞。類似地，CD27表現於天然且活化之效應T細胞以及NK及活化之B細胞兩者上。然而，CD70亦異常表現於各種血液癌，包括急性骨髓白血病(AML)、骨髓發育不良症候群(MDS)及非霍奇金氏淋巴瘤(NHL)以及癌瘤中，且在腫瘤細胞存活及/或腫瘤免疫逃避兩者中起作用。SEA-CD70 (其包含分別SEQ ID NO: 41及42之VH及VL及SEQ ID NO: 53至58之CDR)，經由阻斷CD70/CD27軸傳信，引起抗體依賴性細胞噬菌作用(ADCP)及補體依賴性細胞毒性(CDC)且增強抗體依賴性細胞細胞毒性(ADCC)來起作用。如下文所描述，在皮下NHL模型中測試SEA-CD70與本妥昔單抗維多汀(BV、SGN-35、cAC10-MMAE)之組合。亦稱為SGN-35之本妥昔單抗維多汀為靶向CD30之ADC，其含有與單株抗體cAC10結合之MMAE。CD30表現於霍奇金氏淋巴瘤以及NHL患者子集中。 20.1 物質與方法 NHL 異種移植模型中之皮下腫瘤生長的活體內評估 SEA-CD70 (SEA-h1F6) is an unfucosylated antibody targeting the CD70 antigen. The CD70 molecule is a member of the tumor necrosis factor (TNF) ligand superfamily (TNFSF) and it binds to the related receptor CD27 (TNFRSF7). The interaction between the two molecules activates intracellular signals from both receptors. Under normal conditions, CD70 is transient and restricted to activated T and B cells, mature dendritic cells and natural killer (NK) cells. Similarly, CD27 is expressed on both naive and activated effector T cells as well as NK and activated B cells. However, CD70 is also abnormally expressed in various blood cancers, including acute myeloid leukemia (AML), myelodysplastic syndrome (MDS) and non-Hodgkin's lymphoma (NHL), as well as carcinomas, and is involved in tumor cell survival and/or Tumour immune evasion plays a role in both. SEA-CD70, which comprises VH and VL of SEQ ID NOs: 41 and 42, and CDRs of SEQ ID NOs: 53 to 58, respectively, causes antibody-dependent cellular phagocytosis (ADCP) by blocking the CD70/CD27 axis signaling ) and complement-dependent cytotoxicity (CDC) and enhanced antibody-dependent cellular cytotoxicity (ADCC). The combination of SEA-CD70 and Bentuximab vedotin (BV, SGN-35, cAC10-MMAE) was tested in a subcutaneous NHL model as described below. Bentuximab vedotin, also known as SGN-35, is a CD30-targeted ADC that contains MMAE conjugated to the monoclonal antibody cAC10. CD30 is expressed in Hodgkin's lymphoma and a subset of NHL patients. 20.1 Materials and Methods In vivo assessment of subcutaneous tumor growth in NHL xenograft models

使Farage細胞(2.5×10 6個細胞/動物)再懸浮於0.1 mL 25%基質膠中且皮下注射至SCID小鼠中，該等細胞含有活性先天免疫效應細胞以介導ADCP及ADCC。當達至100 mm 3平均腫瘤大小(藉由使用式：體積(mm 3)=0.5*長度*寬度 2量測，其中長度為較長尺寸)時，將小鼠隨機分為6隻小鼠/組之處理組。腹膜內給予處理。將原液濃度之抗體及化學療法稀釋至適當濃度且以10微升/公克體重注射至動物中。在整個研究期間每週量測腫瘤長度及寬度以及動物體重至少兩次。植入後十九天，以3 mg/kg SEA-CD70及/或1 mg/kg SGN-35開始給藥。每4天腹膜內(IP)給藥SEA-CD70，持續5次且在第19天IP給藥SGN-35一次追蹤動物直至量測到腫瘤體積超過500 mm 3，此時使動物安樂死。由於腫瘤傾向於在較大尺寸下變成潰瘍，因此選擇500 mm 3為腫瘤尺寸終點。 20.2 結果 Farage cells (2.5 x 106 cells/animal) containing active innate immune effector cells to mediate ADCP and ADCC were resuspended in 0.1 mL of 25% Matrigel and injected subcutaneously into SCID mice. When reaching a mean tumor size of 100 mm3 (measured by using the formula : volume ( mm3 )=0.5*length*width2, where length is the longer dimension), mice were randomly divided into 6 mice/ The treatment group of the group. Treatment was administered intraperitoneally. Stock concentrations of antibody and chemotherapy were diluted to appropriate concentrations and injected into animals at 10 microliters/gram body weight. Tumor length and width and animal body weight were measured at least twice a week throughout the study. Nineteen days after implantation, dosing was initiated with 3 mg/kg SEA-CD70 and/or 1 mg/kg SGN-35. SEA-CD70 was administered intraperitoneally (IP) every 4 days for 5 times and SGN-35 was administered once IP on day 19. Animals were followed until tumor volume exceeding 500 mm3 was measured, at which point the animals were euthanized. Since tumors tend to become ulcerated at larger sizes, 500 mm was chosen as the tumor size endpoint. 20.2 Results

如圖20A中所示，與單一藥劑SGN-35或SEA-CD70處理相比，將SGN-35與SEA-CD70組合會延遲腫瘤生長。在第28.5天(處理後第9.5天)，所有未經處理腫瘤之體積增大超過其原始尺寸5倍，而分別在第34.5天及第46天(處理後第15.5天及第27天)，用單一藥劑SEA-CD70或單一藥劑SGN-35處理之腫瘤達至平均5倍增加。尤其，在實驗推斷時(第50天)，用SEA-CD70及SGN-35的組合處理之腫瘤中無一者達至已確定的尺寸終點(圖20B)。與單一SEA-CD70或SGN-35處理相比，當將SEA-CD70與SGN-35組合時，未觀測到明顯毒性或額外重量損失。此等資料指示攜載MMAE有效負載(SGN-35)之ADC與未經岩藻糖基化抗體(SEA-CD70)的組合為有效且極耐受的。 實例 21 ： 未經岩藻糖基化 SEA-BCMA 抗體與奧瑞他汀 ADC 的組合之協同效果 As shown in Figure 20A, combining SGN-35 with SEA-CD70 delayed tumor growth compared to single agent SGN-35 or SEA-CD70 treatment. On day 28.5 (day 9.5 post-treatment), the volume of all untreated tumors increased by 5-fold over their original size, while at day 34.5 and day 46 (day 15.5 and day 27 post-treatment), respectively, Tumors treated with either single agent SEA-CD70 or single agent SGN-35 achieved a mean 5-fold increase. In particular, at experimental extrapolation (day 50), none of the tumors treated with the combination of SEA-CD70 and SGN-35 reached the established size endpoint (FIG. 20B). No significant toxicity or additional weight loss was observed when SEA-CD70 was combined with SGN-35 compared to SEA-CD70 or SGN-35 treatment alone. These data indicate that the combination of the ADC carrying the MMAE payload (SGN-35) and the non-fucosylated antibody (SEA-CD70) is potent and well tolerated. Example 21 : Synergistic effect of combination of non-fucosylated SEA-BCMA antibody and auristatin ADC

SEA-BCMA為靶向B細胞成熟抗原(BCMA)之未經岩藻糖基化抗體，其表現於多發性骨髓瘤(MM)上。SEA-BCMA (其分別具有SEQ ID NO: 45及46之VH及VL，及SEQ ID NO: 47至52之CDR)，經由阻斷配位體介導之BCMA細胞傳信、抗體依賴性細胞噬菌作用(ADCP)及增強型抗體依賴性細胞毒性(ADCC)起作用。如下所述，在散播性MM腫瘤異種移植模型中測試SEA-BCMA與SGN-CD48A之組合。SGN-CD48A為含有葡萄糖苷酸鍵聯之MMAE的靶向CD48之ADC。CD48廣泛表現於MM中。 21.1 物質與方法 21.1.1異種移植模型之活體內存活率評估 SEA-BCMA is an unfucosylated antibody targeting B cell maturation antigen (BCMA), which is expressed in multiple myeloma (MM). SEA-BCMA (which has VH and VL of SEQ ID NOs: 45 and 46, respectively, and CDRs of SEQ ID NOs: 47 to 52), via blocking ligand-mediated BCMA cell signaling, antibody-dependent cytophagy Bacterial effect (ADCP) and enhanced antibody-dependent cytotoxicity (ADCC) play a role. The combination of SEA-BCMA and SGN-CD48A was tested in a disseminated MM tumor xenograft model as described below. SGN-CD48A is a CD48-targeting ADC containing glucuronide-linked MMAE. CD48 is widely expressed in MM. 21.1 Substances and methods 21.1.1 In vivo survival assessment of xenograft models

將MM1S MM細胞IV注射至SCID動物中，該等細胞含有活性先天免疫效應細胞以介導ADCP及ADCC。植入後七天，以0.1 mg/kg SEA-BCMA及/或0.01 mg/kg SGN-CD48A開始給藥，且監測動物存活率。每週IP給藥SEA-BCMA一次，持續5週且IP給藥SGN-CD48A一次。追蹤動物160天之存活率(N=8/組)。截至第51天，所有未處理動物根據IACUC協定經人道地安樂死。 21.1.2異種移植模型之活體內螢光素酶評估 MM1S MM cells, which contain active innate immune effector cells to mediate ADCP and ADCC, were injected IV into SCID animals. Seven days after implantation, dosing was started with 0.1 mg/kg SEA-BCMA and/or 0.01 mg/kg SGN-CD48A, and animal survival was monitored. SEA-BCMA was administered IP once weekly for 5 weeks and SGN-CD48A was administered IP once. Animals were followed for 160 days of survival (N=8/group). By day 51, all untreated animals were humanely euthanized according to IACUC protocol. 21.1.2 In vivo luciferase assessment in xenograft models

將L363螢光素酶MM細胞IV注射至SCID動物中且使MM細胞回歸至骨髓。隨時間監測螢光素酶信號。植入後三十天，以3 mg/kg SEA-BCMA及/或0.3 mg/kg SGN-CD48A開始給藥。每週IP給藥SEA-BCMA一次，持續5週且IP給藥SGN-CD48A一次。追蹤動物175天(N=5/組)。截至第58天，所有未處理動物根據IACUC協定經人道地安樂死。 21.2 結果 L363 luciferase MM cells were injected IV into SCID animals and the MM cells were allowed to return to the bone marrow. The luciferase signal was monitored over time. Thirty days after implantation, dosing was initiated with 3 mg/kg SEA-BCMA and/or 0.3 mg/kg SGN-CD48A. SEA-BCMA was administered IP once weekly for 5 weeks and SGN-CD48A was administered IP once. Animals were followed for 175 days (N=5/group). By day 58, all untreated animals were humanely euthanized according to IACUC protocols. 21.2 Results

如圖21A至圖21B中所示，SGN-CD48A與SEA-BCMA之組合在所測試小鼠模型中誘導完全緩解且延長存活期。在圖21A中，截至第160天，與用SEA-BCMA單獨處理之零動物及用SGN-CD8A單獨處理之一隻動物相比，接受組合療法之組中之八隻動物中有五隻仍活著。在圖21B中，截至第37天，用組合療法處理之所有動物均顯示沒有可偵測的螢光素酶信號，直至第175天研究結束，該信號仍不存在。此驚人的協同作用可歸因於誘導ICD之奧瑞他汀ADC與結合先天免疫細胞之SEA-BCMA的獨特組合。 實例 22 ： 維多汀 ADC 活體內誘導免疫細胞募集及活化 22.1 物質與方法 As shown in Figures 21A-21B, the combination of SGN-CD48A and SEA-BCMA induced complete remission and prolonged survival in the mouse models tested. In Figure 21A, as of day 160, five of the eight animals in the group receiving the combination therapy were still alive compared to zero animals treated with SEA-BCMA alone and one animal treated with SGN-CD8A alone on. In Figure 21B, all animals treated with the combination therapy showed no detectable luciferase signal by day 37, which remained absent until the end of the study on day 175. This surprising synergy can be attributed to the unique combination of ICD-inducing auristatin ADC and SEA-BCMA binding to innate immune cells. Example 22 : Vidotin ADC Induces Immune Cell Recruitment and Activation In Vivo 22.1 Substances and Methods

自用vc-MMAE ADC或未結合vc-MMAE同種型ADC處理8天之動物分離腫瘤異種移植且進行流式細胞量測術或細胞介素分析。CD45陽性免疫細胞經CD11c染色且藉由染色細胞表面上之MHC II類表現而觀測到活化。藉由Luminex量測瘤內細胞介素。 22.2 結果 Tumor xenografts were isolated from animals treated with vc-MMAE ADCs or unconjugated vc-MMAE isotype ADCs for 8 days and subjected to flow cytometry or interferon analysis. CD45 positive immune cells were stained with CD11c and activation was observed by MHC class II expression on the stained cell surface. Intratumoral cytokines were measured by Luminex. 22.2 Results

如圖22中所示，用靶向常見腫瘤抗原之基於MMAE之ADC (vc-MMAE ADC)處理腫瘤攜帶小鼠使得促進腫瘤中之免疫細胞募集及活化。與非結合對照(非結合ADC)相比，當用靶向腫瘤之基於MMAE之ADC (vcMMAE ADC)處理時，樹突狀細胞浸潤及樹突狀細胞抗原呈遞均顯著增強(圖22B)。當用靶向腫瘤之基於MMAE之ADC (vc-MMAE ADC)處理時，瘤內細胞介素含量亦顯著提高(圖22C)。此等資料表明包含微管蛋白干擾劑之ADC以使得促進腫瘤中免疫細胞募集及活化的方式誘導ER應激及腫瘤細胞死亡。As shown in Figure 22, treatment of tumor-bearing mice with an MMAE-based ADC targeting common tumor antigens (vc-MMAE ADC) resulted in enhanced immune cell recruitment and activation in the tumor. Both dendritic cell infiltration and dendritic cell antigen presentation were significantly enhanced when treated with a tumor-targeted MMAE-based ADC (vcMMAE ADC) compared to a non-binding control (non-binding ADC) (Figure 22B). Intratumoral interleukin levels were also significantly increased when treated with a tumor-targeted MMAE-based ADC (vc-MMAE ADC) (FIG. 22C). These data suggest that ADCs comprising tubulin interfering agents induce ER stress and tumor cell death in a manner that promotes the recruitment and activation of immune cells in tumors.

此等結果表明基於MMAE之ADC為用於免疫檢查點阻斷劑之較佳搭配物。 實例 23 ： 藉由維多汀 ADC 誘導 T 細胞 記憶 23.1 物質與方法 These results suggest that MMAE-based ADCs are better partners for immune checkpoint blockade. Example 23 : Induction of T cell memory by vedotin ADC 23.1 Substances and methods

Balb/c小鼠在第0天經皮下在側腹植入表現腫瘤抗原EphA2之Renca同基因型腫瘤細胞株。當達至100 mm 3平均腫瘤大小(藉由使用公式：體積(mm 3)=0.5*長度*寬度 2量測，其中長度為較長尺寸)時，將小鼠隨機分為5隻小鼠/組之處理組。接著，動物用指定的腹膜內處理進行處理(圖21A)。給予各處理一次。將原液濃度之抗體稀釋至適當濃度且以100 µL體積注射至動物體內。針對靶向腫瘤之ADC (ADC-vcMMAE)及非結合ADC (同種型vcMMAE，亦稱為h00-vcMMAE)之最終劑量為5 mg/kg。在整個研究期間量測腫瘤長度、腫瘤寬度及小鼠重量，且使用上式計算腫瘤體積。追蹤動物直至腫瘤體積達至約1,000 mm 3，此時使動物安樂死。 Balb/c mice were implanted subcutaneously on day 0 with Renca isogenic tumor cell lines expressing the tumor antigen EphA2 in the flanks. Mice were randomly divided into 5 mice/ The treatment group of the group. Next, animals were treated with the indicated intraperitoneal treatments (FIG. 21A). Each treatment was given once. The stock concentration of antibody was diluted to the appropriate concentration and injected into animals in a volume of 100 µL. The final dose of the tumor-targeted ADC (ADC-vcMMAE) and the unconjugated ADC (isotype vcMMAE, also known as h00-vcMMAE) was 5 mg/kg. Tumor length, tumor width, and mouse weight were measured throughout the study, and tumor volume was calculated using the above formula. Animals were followed until tumor volume reached approximately 1,000 mm3 , at which point the animals were euthanized.

監測達成治癒性抗腫瘤反應之小鼠。接著，在達成治癒之後30天，用Renca腫瘤細胞再攻擊小鼠(圖21B)，且評估新腫瘤之過度生長及排斥。 23.2 結果 Mice that achieved a curative antitumor response were monitored. Then, 30 days after cure was achieved, mice were rechallenged with Renca tumor cells (FIG. 21B), and new tumors were assessed for overgrowth and rejection. 23.2 Results

如圖23A中所示，在Renca基因型模型中，用單一劑量之靶向腫瘤的MMAE ADC (ADC-vcMMAE)進行之處理引起強抗腫瘤活性及治癒反應。如圖23B中所示，當藉由MMAE ADC處理治癒之小鼠經Renca腫瘤細胞再攻擊以評估免疫記憶之誘導時，此類小鼠能夠排斥隨後移植之腫瘤細胞。此類結果展現出MMAE ADC能夠引發特定抗腫瘤T細胞反應。 實例 24 ：經 本妥昔單抗維多汀 (BV ； SGN-35) 處理之細胞賦予保護性抗腫瘤免疫 24.1 物質與方法 As shown in Figure 23A, treatment with a single dose of tumor-targeted MMAE ADC (ADC-vcMMAE) resulted in strong antitumor activity and a curative response in the Renca genotype model. As shown in Figure 23B, when mice cured by MMAE ADC treatment were rechallenged with Renca tumor cells to assess induction of immune memory, such mice were able to reject subsequently transplanted tumor cells. Such results demonstrate that MMAE ADCs can elicit specific anti-tumor T cell responses. Example 24 : Bentuximab vedotin (BV ; SGN-35) -treated cells confers protective antitumor immunity 24.1 Substances and methods

表現人類CD30之A20細胞用CD30-奧瑞他汀ADC (BV；SGN-35)或MMAE處理18小時。替代地，將細胞之一個等分試樣急驟冷凍。執行經處理樣本之Ficoll離心以移除活細胞。藉由流式細胞量測術使用膜聯蛋白V/7AAD分析所有樣本之細胞凋亡及存活率。將正在死亡及死亡細胞洗滌，再懸浮於PBS中且經腹膜內注射至小鼠體內。小鼠經免疫接種2次，間隔7天。將免疫接種小鼠再靜置7天，且接著用A20淋巴瘤細胞攻擊，且隨時間監測腫瘤生長或排斥。 24.2 結果 A20 cells expressing human CD30 were treated with CD30-auristatin ADC (BV; SGN-35) or MMAE for 18 hours. Alternatively, one aliquot of cells was snap frozen. Ficoll centrifugation of the treated samples was performed to remove viable cells. All samples were analyzed for apoptosis and viability by flow cytometry using Annexin V/7AAD. Dying and dead cells were washed, resuspended in PBS and injected intraperitoneally into mice. Mice were immunized twice, 7 days apart. The immunized mice were left undisturbed for an additional 7 days and then challenged with A20 lymphoma cells and monitored over time for tumor growth or rejection. 24.2 Results

如圖24中所示，與免疫接種有經急驟冷凍殺死(細胞死亡之非ICD方法)之表現CD30之A20細胞的小鼠相比，免疫接種有使用BV或MMAE殺死之表現CD30的A20細胞之小鼠呈現排斥移植A20細胞之更強免疫反應。此等結果指示記憶T細胞反應之誘導。免疫記憶之誘導被視為評估分子之ICD活性的最高準則。As shown in Figure 24, immunization with A20 expressing CD30 killed using BV or MMAE compared mice immunized with A20 expressing CD30 cells killed by flash freezing (a non-ICD method of cell death) Cell-derived mice exhibited stronger immune responses that rejected the transplanted A20 cells. These results are indicative of induction of memory T cell responses. Induction of immune memory is considered the highest criterion for assessing the ICD activity of molecules.

總體而言，前述實例中呈現之結果支持基於奧瑞他汀之ADC (例如MMAE及MMAF)誘導免疫原性細胞死亡的獨特能力。如藉由前述實例所表明，奧瑞他汀之作用機制及其破壞微管網路之能力似乎與引起危險信號(DAMP)之暴露及分泌的ER應激反應之誘導相關聯。此等DAMP之暴露引發先天免疫細胞反應，其可引起抗原特異性T細胞反應。誘導可識別腫瘤抗原之新抗原特異性T細胞可在臨床前引起與提供長期免疫保護之長期記憶T細胞反應相關聯的治癒性抗腫瘤活性。Overall, the results presented in the preceding examples support the unique ability of auristatin-based ADCs such as MMAE and MMAF to induce immunogenic cell death. As demonstrated by the foregoing examples, the mechanism of action of auristatin and its ability to disrupt the microtubule network appears to be related to the induction of ER stress responses that lead to exposure to danger signals (DAMPs) and secretion. Exposure to these DAMPs elicits innate immune cell responses that can elicit antigen-specific T cell responses. Induction of neoantigen-specific T cells that recognize tumor antigens can elicit preclinical curative antitumor activity associated with long-term memory T cell responses that provide long-term immune protection.

由MMAE ADC誘導之此記憶T細胞群可藉由未經岩藻糖基化抗體進一步增加及/或增強。在確定由誘導ICD之MMAE ADC產生的免疫記憶力(例如經由上述記憶T細胞反應)後，諸如SEA-TIGIT之未經岩藻糖基化抗體可經由與PD-1/PD-L1之檢查點抑制性機制類似的腫瘤阻斷/抑制性機制進一步擴增免疫反應。This memory T cell population induced by MMAE ADCs can be further increased and/or enhanced by non-fucosylated antibodies. After determining the immune memory produced by ICD-inducing MMAE ADCs (eg, via the memory T cell responses described above), non-fucosylated antibodies such as SEA-TIGIT can be inhibited via checkpoints with PD-1/PD-L1 A tumor-blocking/suppressive mechanism similar to the sexual mechanism further amplifies the immune response.

此外，將基於奧瑞他汀之ADC (例如MMAE及MMAF)，諸如MMAE ADC驅動免疫原性細胞死亡之能力與免疫細胞促效作用配對可擴增抗腫瘤活性。免疫促效作用可藉由使用未經岩藻糖基化抗體或已經工程化以增強與活化FcγR之結合及/或降低與抑制性FcγR之結合的抗體來擴增(例如如上述實例關於未經岩藻糖基化CD40及BCMA抗體所展示)。未經岩藻糖基化抗體可增加與活化FcγRIIIa受體之結合及降低或最小化與抑制性FcγRIIb受體之結合。視抗體目標之性質而定，此屬性為多模式的。在如同CD40之受體在聚集時具有最佳活性的情況下，結合於FcγRIIA+細胞之未經岩藻糖基化抗體增強受體集群以及免疫促效作用及活化。 參見圖25。如在TIGIT之情況下，未經岩藻糖基化抗體增加抗原(+) T細胞與抗原呈遞細胞之間的免疫突觸強度(圖25)。先天細胞上之FcγRIIIa接合增強其活化且產生可增強抗原特異性T細胞反應之因子。最後，未經岩藻糖基化主鏈可獨立於目標抗原而結合於先天免疫細胞或諸如γ δ T細胞之其他FcγRIIIa細胞，以誘導可幫助引發二級抗原特異性T細胞反應之活化狀態。未經岩藻糖基化抗體藉以工作之所有此等機制可引起驅動抗腫瘤活性且長期存在的免疫保護之T細胞反應。降低或缺少於FcγRIIb之結合意謂不存在降低由未經岩藻糖基化抗體驅動之免疫活化的計數或抑制性信號。 In addition, pairing the ability of auristatin-based ADCs (eg, MMAE and MMAF), such as MMAE ADCs, to drive immunogenic cell death with immune cell agonism can amplify antitumor activity. The immunostimulatory effect can be amplified by using unfucosylated antibodies or antibodies that have been engineered to enhance binding to activating FcyRs and/or reduce binding to inhibitory FcyRs (e.g., as described above in the examples for non-fucosylated FcyRs). Fucosylated CD40 and BCMA antibodies displayed). Unfucosylated antibodies can increase binding to activating FcγRIIIa receptors and decrease or minimize binding to inhibitory FcγRIIb receptors. Depending on the nature of the antibody target, this property is multimodal. Where receptors like CD40 have optimal activity upon aggregation, non-fucosylated antibodies bound to FcγRIIA+ cells enhanced receptor aggregation as well as immunostimulatory effects and activation. See Figure 25. As in the case of TIGIT, unfucosylated antibodies increased the strength of immune synapses between antigen (+) T cells and antigen presenting cells (Figure 25). FcyRIIIa engagement on innate cells enhances their activation and produces factors that enhance antigen-specific T cell responses. Finally, the unfucosylated backbone can bind to innate immune cells or other FcγRIIIa cells such as γδ T cells independently of the target antigen to induce an activated state that can help elicit secondary antigen-specific T cell responses. All of these mechanisms by which unfucosylated antibodies work can elicit T cell responses that drive antitumor activity and long-lasting immune protection. Reduced or absent binding to FcyRIIb means that there is no count or inhibitory signal that reduces immune activation driven by the non-fucosylated antibody.

諸如MMAE ADC之奧瑞他汀ADC的作用機制藉由未經岩藻糖基化mAb與免疫調節偶合產生協同性且互補活性，其展示引起增強的免疫活化及治癒性抗腫瘤反應，如本文中所表明。The mechanism of action of auristatin ADCs such as MMAE ADCs produces synergistic and complementary activities by coupling non-fucosylated mAbs to immunomodulatory activities, which have been shown to cause enhanced immune activation and curative antitumor responses, as described herein. show.

本文中中所引用之所有公開案、專利、專利申請案及其他文獻皆出於所有目的特此以全文引用之方式併入，引用的程度就如同個別地指示將各個別公開案、專利案、專利申請案及其他文獻以引用之方式併入以用於所有目的一樣。 序列表 名稱 SEQ ID NO 序列 抗TIGIT抗體純系13 VH蛋白質 1 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGSIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSS 抗TIGIT抗體純系13A VH 2 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSLIPYFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSS 抗TIGIT抗體純系13B VH 3 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAWAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVSGILGYVWFDPWGQGTLVTVSS 抗TIGIT抗體純系13C VH 4 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPLFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSS 抗TIGIT抗體純系13D VH 5 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSS 純系13、13A、13B、13C及13D VL蛋白質 6 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQARRIPITFGGGTKVEIK 純系13 VH CDR1 7 GTFSSYAIS 純系13A、13C及13D VH CDR1 8 GTFLSSAIS 純系13B VH CDR1 9 GTFSAWAIS 純系13 VH CDR2 10 SIIPIFGTANYAQKFQG 純系13A VH CDR2 11 SLIPYFGTANYAQKFQG 純系13B及13D VH CDR2 12 SIIPYFGKANYAQKFQG 純系13C VH CDR2 13 SIIPLFGKANYAQKFQG 純系13及13A VH CDR3 14 ARGPSEVGAILGYVWFDP 純系13B VH CDR3 15 ARGPSEVSGILGYVWFDP 純系13C及3D VH CDR3 16 ARGPSEVKGILGYVWFDP 純系13、13A、13B、13C及13D VL CDR1 17 RSSQSLLHSNGYNYLD 純系13、13A、13B、13C及13D VL CDR2 18 LGSNRAS 純系13、13A、13B、13C及13D VL CDR3 19 MQARRIPIT 純系13重鏈hIgG1 (及未經岩藻糖基化hIgG1)胺基酸序列；粗體指示VH；SEA-TGT 20 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGSIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 純系13A重鏈hIgG1 (及未經岩藻糖基化hIgG1)胺基酸序列；粗體指示VH 21 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSLIPYFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 純系13B重鏈hIgG1 (及未經岩藻糖基化hIgG1)胺基酸序列；粗體指示VH 22 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAWAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVSGILGYVWFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 純系13C重鏈hIgG1 (及未經岩藻糖基化hIgG1)胺基酸序列；粗體指示VH 23 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPLFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 純系13D重鏈hIgG1 (及未經岩藻糖基化hIgG1)胺基酸序列；粗體指示VH 24 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 純系13、13A、13B、13C及13D輕鏈hκ (及未經岩藻糖基化)胺基酸序列；粗體指示VL；SEA-TGT 25 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQARRIPITFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEA-CD40 (未經岩藻糖基化hS2C6)重鏈 26 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYYIHWVRQAPGKGLEWVARVIPNAGGTSYNQKFKGRFTLSVDNSKNTAYLQMNSLRAEDTAVYYCAREGIYWWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEA-CD40 (未經岩藻糖基化hS2C6)輕鏈 27 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGNTFLHWYQQKPGKAPKLLIYTVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCSQTTHVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEA-CD40之VH 28 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYYIHWVRQAPGKGLEWVARVIPNAGGTSYNQKFKGRFTLSVDNSKNTAYLQMNSLRAEDTAVYYCAREGIYWWGQGTLVTVSS SEA-CD40之VL 29 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGNTFLHWYQQKPGKAPKLLIYTVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCSQTTHVPWTFGQGTKVEIK SEA-CD40 VH CDR1 30 GYYIH SEA-CD40 VH CDR2 31 RVIPNAGGTSYNQKFKG SEA-CD40 VH CDR3 32 EGIYW SEA-CD40 VL CDR1 33 RSSQSLVHSNGNTFLH SEA-CD40 VL CDR2 34 TVSNRFS SEA-CD40 VL CDR3 35 SQTTHVPWT 替代抗CD40抗體CDR2 36 RVIPQAGGTSYNQKFKG SGN-B6A重鏈可變區 37 QFQLVQSGAEVKKPGASVKVSCKASGYSFTDYNVNWVRQAPGQGLEWIGVINPKYGTTRYNQKFKGRATLTVDKSTSTAYMELSSLRSEDTAVYYCTRGLNAWDYWGQGTLVTVSS SGN-B6A輕鏈可變區 38 DIQMTQSPSSLSASVGDRVTITCGASENIYGALNWYQQKPGKAPKLLIYGATNLEDGVPSRFSGSGSGRDYTFTISSLQPEDIATYYCQNVLTTPYTFGQGTKLEIK SGN-STNV重鏈可變區 39 EVQLVQSGAEVKKPGASVKVSCKASGYTFTDHAIHWVRQAPGQGLEWMGYFSPGNDDIKYNEKFRGRVTMTADKSSSTAYMELRSLRSDDTAVYFCKRSLSTPYWGQGTLVTVSS SGN-STNV重鏈可變區 40 DIVMTQSPDSLAVSLGERATINCKSSQSLLNRGNHKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYTYPYTFGQGTKVEIK SEA-CD70重鏈可變區 41 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLKWMGWINTYTGEPTYADAFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDYGDYGMDYWGQGTTVTVSS SEA-CD70輕鏈可變區 42 DIVMTQSPDSLAVSLGERATINCRASKSVSTSGYSFMHWYQQKPGQPPKLLIYLASNLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQHSREVPWTFGQGTKVEIK SGN-CD228A重鏈可變區 43

Figure 02_image021
SGN-CD228A輕鏈可變區 44
Figure 02_image023
 SGN-BCMA重鏈可變區 45 QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYIHWVRQAPGQGLEWIGYINPNSGYTNYAQKFQGRATMTADKSINTAYVELSRLRSDDTAVYFCTRYMWERVTGFFDFWGQGTMVTVSS SGN-BCMA輕鏈可變區 46 DIQMTQSPSSVSASVGDRVTITCLASEDISDDLAWYQQKPGKAPKVLVYTTSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQTYKFPPTFGGGTKVEIK SGN-BCMA VH CDR1 47 DYYIH SGN-BCMA VH CDR2 48 YINPNSGYTNYAQKFQG SGN-BCMA VH CDR3 49 YMWERVTGFFDF SGN-BCMA VL CDR1 50 LASEDISDDLA SGN-BCMA VL CDR2 51 TTSSLQS SGN-BCMA VL CDR3 52 QQTYKFPPT SEA-CD70 VH CDR1 53 NYGMN SEA-CD70 VH CDR2 54 WINTYTGEPTYADAFKG SEA-CD70 VH CDR3 55 DYGDYGMDY SEA-CD70 VL CDR1 56 RASKSVSTSGYSFMH SEA-CD70 VL CDR2 57 LASNLES SEA-CD70 VL CDR3 58 QHSREVPWT 唑貝妥西單抗(175D10)重鏈可變區 59 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSS 唑貝妥西單抗(175D10)輕鏈可變區 60 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTK 唑貝妥西單抗(175D10) VH CDR1 61 SYWIN 唑貝妥西單抗(175D10) VH CDR2 62 NIYPSDSYTNYNQKFKD 唑貝妥西單抗(175D10) VH CDR3 63 SWRGNSFDY 唑貝妥西單抗(175D10) VL CDR1 64 KSSQSLLNSGNQKNYLT 唑貝妥西單抗(175D10) VL CDR2 65 WASTRES 唑貝妥西單抗(175D10) VL CDR3 66 QNDYSYPFT 163E12重鏈可變區 67 QIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNWVKQAPGKGLKWMGWINTNTGEPTYAEEFKGRFAFSLETSASTAYLQINNLKNEDTATYFCARLGFGNAMDYWGQGTSVTVSS 163E12輕鏈可變區 68 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLELK 163E12 VH CDR1 69 NYGMN 163E12 VH CDR2 70 WINTNTGEPTYAEEFKG 163E12 VH CDR3 71 LGFGNAMDY 163E12 VL CDR1 72 KSSQSLLNSGNQKNYLT 163E12 VL CDR2 73 WASTRES 163E12 VL CDR3 74 QNDYSYPLT SGN-PDL1V重鏈可變區 75 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTAAISWVRQAPGQGLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYFCARKFHFVSGSPFGMDVWGQGTTVTVSS SGN-PDL1V輕鏈可變區 76 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPTFGQGTKVEIK SGN-PDL1V VH CDR1 77 TAAIS SGN-PDL1V VH CDR2 78 GIIPIFGKAHYAQKFQG SGN-PDL1V VH CDR3 79 KFHFVSGSPFGMDV SGN-PDL1V VL CDR1 80 RASQSVSSYLA SGN-PDL1V VL CDR2 81 DASNRAT SGN-PDL1V VL CDR3 82 QQRSNWPT SGN-ALPV重鏈可變區 83 EVQLVESGGGLVQPGRSLRLSCTASGFTFTDYYMSWVRQAPGKGLEWLALIRNKATGYTTEYTASVKGRFTISRDNSKSILYLQMNSLKTEDTAVYYCARASFYYDGKVLAYWGQGTLVTVSS SGN-ALPV輕鏈可變區 84 DTQMTQSPSSLSASVGDRVTITCQASQDINKYLAWYQYKPGKAPKLLIHYTSSLQSGVPSRFSGSGSGRDYTFTISSLQPEDIATYYCLQYDNLYTFGQGTKLEIK SGN-ALPV VH CDR1 85 DYYMS SGN-ALPV VH CDR2 86 LIRNKATGYTTEYTASVKG SGN-ALPV VH CDR3 87 ASFYYDGKVLAY SGN-ALPV VL CDR1 88 QASQDINKYLA SGN-ALPV VL CDR2 89 YTSSLQS SGN-ALPV VL CDR3 90 LQYDNLYT SGN-B7H4V重鏈可變區 91 QLQLQESGPGLVKPSETLSLTCTVSGGSIKSGSYYWGWIRQPPGKGLEWIGNIYYSGSTYYNPSLRSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGSYPNQFDPWGQGTLVTVSS SGN-B7H4V輕鏈可變區 92 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYHSFPFTFGGGTKVEIK SGN-B7H4V VH CDR1 93 GSIKSGSYYWG SGN-B7H4V VH CDR2 94 NIYYSGSTYYNPSLRS SGN-B7H4V VH CDR3 95 AREGSYPNQFDP SGN-B7H4V VL CDR1 96 RASQSVSSNLA SGN-B7H4V VL CDR2 97 GASTRAT SGN-B7H4V VL CDR3 98 QQYHSFPFT 迪西妥單抗維多汀重鏈 99 EVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYIHWVQQAPGKGLEWMGRVNPDHGDSYYNQKFKDKATITADKSTDTAYMELSSLRSEDTAVYFCARNYLFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 迪西妥單抗維多汀輕鏈 100 DIQMTQSPSSVSASVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASIRHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQFATYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 立伐土珠單抗維多汀重鏈 101 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVATIGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHRGFDVGHFDFWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 立伐土珠單抗維多汀輕鏈 102 DIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGKAPKLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSFNPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 恩諾單抗維多汀(EV)重鏈可變區 103 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYNMNWVRQAPGKGLEWVSYISSSSSTIYYADSVKGRFTISRDNAKNSLSLQMNSLRDEDTAVYYCARAYYYGMDVWGQGTTVTVSS 恩諾單抗維多汀(EV)輕鏈可變區 104 DIQMTQSPSSVSASVGDRVTITCRASQGISGWLAWYQQKPGKAPKFLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPTFGGGTKVEIK 恩諾單抗維多汀(EV) VH CDR1 105 SYNMN 恩諾單抗維多汀(EV) VH CDR2 106 YISSSSSTIYYADSVKG 恩諾單抗維多汀(EV) VH CDR3 107 AYYYGMDV 恩諾單抗維多汀(EV) VL CDR1 108 RASQGISGWLA 恩諾單抗維多汀(EV) VL CDR2 109 AASTLQS 恩諾單抗維多汀(EV) VL CDR3 110 QQANSFPPT h2A2重鏈可變區 111 QFQLVQSGAEVKKPGASVKVSCKASGYSFTDYNVNWVRQAPGQGLEWIGVINPKYGTTRYNQKFKGRATLTVDKSTSTAYMELSSLRSEDTAVYYCTRGLNAWDYWGQGTLVTVSS h2A2輕鏈可變區 112 DIQMTQSPSSLSASVGDRVTITCGASENIYGALNWYQQKPGKAPKLLIYGATNLEDGVPSRFSGSGSGRDYTFTISSLQPEDIATYYCQNVLTTPYTFGQGTKLEIK h2A2 VH CDR1 113 DYNVN h2A2 VH CDR2 114 VINPKYGTTRYNQKFKG h2A2 VH CDR3 115 GLNAWDY h2A2 VL CDR1 116 GASENIYGALN h2A2 VL CDR2 117 GATNLED h2A2 VL CDR3 118 QNVLTTPYT h15H3重鏈可變區 119 QVQLVQSGAEVKKPGASVKVSCKASGYSFSGYFMNWVRQAPGQGLEWMGLINPYNGDSFYNQKFKGRVTMTRQTSTSTVYMELSSLRSEDTAVYYCVRGLRRDFDYWGQGTLVTVSS h15H3輕鏈可變區 120 DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLFQRPGQSPRRLIYLVSELDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPRTFGGGTKLEIK h15H3 VH CDR1 121 GYFMN h15H3 VH CDR2 122 LINPYNGDSFYNQKFKG h15H3 VH CDR3 123 GLRRDFDY h15H3 VL CDR1 124 KSSQSLLDSDGKTYLN h15H3 VL CDR2 125 LVSELDS h15H3 VL CDR3 126 WQGTHFPRT SGN-CD228A重鏈可變區 127 QVQLQESGPGLVKPSETLSLTCTVSGDSITSGYWNWIRQPPGKGLEYIGYISDSGITYYNPSLKSRVTISRDTSKNQYSLKLSSVTAADTAVYYCARRTLATYYAMDYWGQGTLVTVSS SGN-CD228A輕鏈可變區 128 DFVMTQSPLSLPVTLGQPASISCRASQSLVHSDGNTYLHWYQQRPGQSPRLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPPTFGQGTKLEIK SGN-CD228A VH CDR1 129 SGYWN SGN-CD228A VH CDR2 130 YISDSGITYYNPSLKS SGN-CD228A VH CDR3 131 RTLATYYAMDY SGN-CD228A VL CDR1 132 RASQSLVHSDGNTYLH SGN-CD228A VL CDR2 133 RVSNRFS SGN-CD228A VL CDR3 134 SQSTHVPPT SGN-LIV1A拉地妥珠單抗維多汀(LV)重鏈可變區 135 QVQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWMGWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAVHNAHYGTWFAYWGQGTLVTVSS SGN-LIV1A拉地妥珠單抗維多汀(LV)輕鏈可變區 136 DVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEWYQQRPGQSPRPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGGGTKVEIK SGN-LIV1A拉地妥珠單抗維多汀(LV) VH CDR1 137 DYYMH SGN-LIV1A拉地妥珠單抗維多汀(LV) VH CDR2 138 WIDPENGDTEYGPKFQG SGN-LIV1A拉地妥珠單抗維多汀(LV) VH CDR3 139 HNAHYGTWFAY SGN-LIV1A拉地妥珠單抗維多汀(LV) VL CDR1 140 RSSQSLLHSSGNTYLE SGN-LIV1A拉地妥珠單抗維多汀(LV) VL CDR2 141 KISTRFS SGN-LIV1A拉地妥珠單抗維多汀(LV) VL CDR3 142 FQGSHVPYT 替索圖單抗維多汀(TV)重鏈可變區 143 EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGDYTYYTDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPWGYYLDSWGQGTLVTVSS 替索圖單抗維多汀(TV)輕鏈可變區 144 DIQMTQSPPSLSASAGDRVTITCRASQGISSRLAWYQQKPEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKLEIK 替索圖單抗維多汀(TV) VH CDR1 145 GFTFSNYA 替索圖單抗維多汀(TV) VH CDR2 146 ISGSGDYT 替索圖單抗維多汀(TV) VH CDR3 147 ARSPWGYYLDS 替索圖單抗維多汀(TV) VL CDR1 148 QGISSR 替索圖單抗維多汀(TV) VL CDR2 149 AAS 替索圖單抗維多汀(TV) VL CDR3 150 QQYNSYPYT All publications, patents, patent applications, and other documents cited herein are hereby incorporated by reference in their entirety for all purposes to the same extent as if each individual publication, patent, patent The application and other documents are incorporated by reference for all purposes the same. sequence listing name SEQ ID NO sequence Anti-TIGIT antibody clone 13 VH protein 1 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGSIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSS Anti-TIGIT antibody clone 13A VH 2 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSLIPYFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSS Anti-TIGIT antibody clone 13B VH 3 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAWAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVSGILGYVWFDPWGQGTLVTVSS Anti-TIGIT antibody clone 13C VH 4 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPLFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSS Anti-TIGIT antibody clone 13D VH 5 QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSS Pure line 13, 13A, 13B, 13C and 13D VL proteins 6 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQARRIPITFGGGTKVEIK Pure line 13 VH CDR1 7 GTFSSYAIS Pure lines 13A, 13C and 13D VH CDR1 8 GTFLSSAIS Pure line 13B VH CDR1 9 GTFSAWAIS Pure line 13 VH CDR2 10 SIIPIFGTANYAQKFQG Pure line 13A VH CDR2 11 SLIPYFGTANYAQKFQG Pure line 13B and 13D VH CDR2 12 SIIPYFGKANYAQKFQG Pure line 13C VH CDR2 13 SIIPLFGKANYAQKFQG Pure line 13 and 13A VH CDR3 14 ARGPSEVGAILGYVWFDP Pure line 13B VH CDR3 15 ARGPSEVSGILGYVWFDP Pure line 13C and 3D VH CDR3 16 ARGPSEVKGILGYVWFDP Pure lines 13, 13A, 13B, 13C and 13D VL CDR1 17 RSSQSLLHSNGYNYLD Pure lines 13, 13A, 13B, 13C and 13D VL CDR2 18 LGSNRAS Pure lines 13, 13A, 13B, 13C and 13D VL CDR3 19 MQARRIPIT Pure line 13 heavy chain hIgG1 (and unfucosylated hIgG1) amino acid sequence; bold indicates VH; SEA-TGT 20 QVQLVQSGAEVKKPGSSVKVSCKASGGTFSSYAISWVRQAPGQGLEWMGSIIPIFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Pure line 13A heavy chain hIgG1 (and unfucosylated hIgG1) amino acid sequence; bold indicates VH twenty one QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSLIPYFGTANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVGAILGYVWFDPWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Pure line 13B heavy chain hIgG1 (and unfucosylated hIgG1) amino acid sequence; bold indicates VH twenty two QVQLVQSGAEVKKPGSSVKVSCKASGGTFSAWAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVSGILGYVWFDPWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Pure line 13C heavy chain hIgG1 (and unfucosylated hIgG1) amino acid sequence; bold indicates VH twenty three QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPLFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Pure 13D heavy chain hIgG1 (and unfucosylated hIgG1) amino acid sequence; bold indicates VH twenty four QVQLVQSGAEVKKPGSSVKVSCKASGGTFLSSAISWVRQAPGQGLEWMGSIIPYFGKANYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYYCARGPSEVKGILGYVWFDPWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Pure lines 13, 13A, 13B, 13C and 13D light chain hκ (and unfucosylated) amino acid sequences; bold indicates VL; SEA-TGT 25 DIVMTQSPLSLPVTPGEPASISCRSSQSLLHSNGYNYLDWYLQKPGQSPQLLIYLGSNRASGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCMQARRIPITFGGGTKVEIKR TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEA-CD40 (unfucosylated hS2C6) heavy chain 26 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYYIHWVRQAPGKGLEWVARVIPNAGGTSYNQKFKGRFTLSVDNSKNTAYLQMNSLRAEDTAVYYCAREGIYWWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK SEA-CD40 (unfucosylated hS2C6) light chain 27 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGNTFLHWYQQKPGKAPKLLIYTVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCSQTTHVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC VH of SEA-CD40 28 EVQLVESGGGLVQPGGSLRLSCAASGYSFTGYYIHWVRQAPGKGLEWVARVIPNAGGTSYNQKFKGRFTLSVDNSKNTAYLQMNSLRAEDTAVYYCAREGIYWWGQGTLVTVSS VL of SEA-CD40 29 DIQMTQSPSSLSASVGDRVTITCRSSQSLVHSNGNTFLHWYQQKPGKAPKLLIYTVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCSQTTHVPWTFGQGTKVEIK SEA-CD40 VH CDR1 30 GYYIH SEA-CD40 VH CDR2 31 RVIPNAGGTSYNQKFKG SEA-CD40 VH CDR3 32 EGIYW SEA-CD40VL CDR1 33 RSSQSLVHSNGNTFLH SEA-CD40VL CDR2 34 TVSNRFS SEA-CD40VL CDR3 35 SQTTHVPWT Alternative anti-CD40 antibody CDR2 36 RVIPQAGGTSYNQKFKG SGN-B6A heavy chain variable region 37 QFQLVQSGAEVKKPGASVKVSCKASGYSFTDYNVNWVRQAPGQGLEWIGVINPKYGTTRYNQKFKGRATLTVDKSTSTAYMELSSLRSEDTAVYYCTRGLNAWDYWGQGTLVTVSS SGN-B6A light chain variable region 38 DIQMTQSPSSLSASVGDRVTITCGASENIYGALNWYQQKPGKAPKLLIYGATNLEDGVPSRFSGSGSGRDYTFTISSLQPEDIATYYCQNVLTTPYTFGQGTKLEIK SGN-STNV heavy chain variable region 39 EVQLVQSGAEVKKPGASVKVSCKASGYTFTDHAIHWVRQAPGQGLEWMGYFSPGNDDIKYNEKFRGRVTMTADKSSSTAYMELRSLRSDDTAVYFCKRSLSTPYWGQGTLVTVSS SGN-STNV heavy chain variable region 40 DIVMTQSPDSLAVSLGERATINCKSSQSLLNRGNHKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQNDYTYPYTFGQGTKVEIK SEA-CD70 heavy chain variable region 41 QVQLVQSGAEVKKPGASVKVSCKASGYTFTNYGMNWVRQAPGQGLKWMGWINTYTGEPTYADAFKGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDYGDYGMDYWGQGTTVTVSS SEA-CD70 light chain variable region 42 DIVMTQSPDSLAVSLGERATINCRASKSVSTSGYSFMHWYQQKPGQPPKLLIYLASNLESGVPDRFSGSGSGTDFTLTISSLQAEDVAVYYCQHSREVPWTFGQGTKVEIK SGN-CD228A heavy chain variable region 43
Figure 02_image021
 SGN-CD228A light chain variable region 44
Figure 02_image023
 SGN-BCMA heavy chain variable region 45 QVQLVQSGAEVKKPGASVKLSCKASGYTFTDYYIHWVRQAPGQGLEWIGYINPNSGYTNYAQKFQGRATMTADKSINTAYVELSRLRSDDTAVYFCTRYMWERVTGFFDFWGQGTMVTVSS SGN-BCMA light chain variable region 46 DIQMTQSPSSVSASVGDRVTITCLASEDISDDLAWYQQKPGKAPKVLVYTTSSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYFCQQTYKFPPTFGGGTKVEIK SGN-BCMA VH CDR1 47 DYYIH SGN-BCMA VH CDR2 48 YINPNSGYTNYAQKFQG SGN-BCMA VH CDR3 49 YMWERVTGFFDF SGN-BCMA VL CDR1 50 LASEDISDDLA SGN-BCMA VL CDR2 51 TTSSLQS SGN-BCMA VL CDR3 52 QQTYKFPPT SEA-CD70 VH CDR1 53 NYGMN SEA-CD70 VH CDR2 54 WINTYTGEPTYADAFKG SEA-CD70 VH CDR3 55 DYGDYGMDY SEA-CD70VL CDR1 56 RASKSVSTSGYSFMH SEA-CD70VL CDR2 57 LASNLES SEA-CD70VL CDR3 58 QHSREVPWT Zobetuzumab (175D10) heavy chain variable region 59 QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSS Zobetuzumab (175D10) light chain variable region 60 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTK Zobetuzumab (175D10) VH CDR1 61 SYWIN Zobetuzumab (175D10) VH CDR2 62 NIYPSDSYTNYNQKFKD Zobetuzumab (175D10) VH CDR3 63 SWRGNSFDY Zobetuzumab (175D10) VL CDR1 64 KSSQSLLNSGNQKNYLT Zobetuzumab (175D10) VL CDR2 65 WASTRES Zobetuzumab (175D10) VL CDR3 66 QNDYSYPFT 163E12 heavy chain variable region 67 QIQLVQSGPELKKPGETVKISCKASGYTFTNYGMNWVKQAPGKGLKWMGWINTNTGEPTYAEEFKGRFAFSLETSASTAYLQINNLKNEDTATYFCARLGFGNAMDYWGQGTSVTVSS 163E12 light chain variable region 68 DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWASTRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPLTFGAGTKLELK 163E12 VH CDR1 69 NYGMN 163E12 VH CDR2 70 WINTNTGEPTYAEEFKG 163E12 VH CDR3 71 LGFGNAMDY 163E12 VL CDR1 72 KSSQSLLNSGNQKNYLT 163E12 VL CDR2 73 WASTRES 163E12 VL CDR3 74 QNDYSYPLT SGN-PDL1V heavy chain variable region 75 QVQLVQSGAEVKKPGSSVKVSCKTSGDTFSTAAISWVRQAPGQGLEWMGGIIPIFGKAHYAQKFQGRVTITADESTSTAYMELSSLRSEDTAVYFCARKFHFVSGSPFGMDVWGQGTTVTVSS SGN-PDL1V light chain variable region 76 EIVLTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPTFGQGTKVEIK SGN-PDL1V VH CDR1 77 TAAIS SGN-PDL1V VH CDR2 78 GIIPIFGKAHYAQKFQG SGN-PDL1V VH CDR3 79 KFHFVSGSPFGMDV SGN-PDL1V VL CDR1 80 RASQSVSSYLA SGN-PDL1V VL CDR2 81 DASNRAT SGN-PDL1V VL CDR3 82 QQRSNWPT SGN-ALPV heavy chain variable region 83 EVQLVESGGGLVQPGRSLRLSCTASGFTFTDYYMSWVRQAPGKGLEWLALIRNKATGYTTEYTASVKGRFTISRDNSKSILYLQMNSLKTEDTAVYYCARASFYYDGKVLAYWGQGTLVTVSS SGN-ALPV light chain variable region 84 DTQMTQSPSSLSASVGDRVTITCQASQDINKYLAWYQYKPGKAPKLLIHYTSSLQSGVPSRFSGSGSGRDYTFTISSLQPEDIATYYCLQYDNLYTFGQGTKLEIK SGN-ALPV VH CDR1 85 DYYMS SGN-ALPV VH CDR2 86 LIRNKATGYTTEYTASVKG SGN-ALPV VH CDR3 87 ASFYYDGKVLAY SGN-ALPV VL CDR1 88 QASQDINKYLA SGN-ALPV VL CDR2 89 YTSSLQS SGN-ALPV VL CDR3 90 LQYDNLYT SGN-B7H4V heavy chain variable region 91 QLQLQESGPGLVKPSETLSLTCTVSGGSIKSGSYYWGWIRQPPGKGLEWIGNIYYSGSTYYNPSLRSRVTISVDTSKNQFSLKLSSVTAADTAVYYCAREGSYPNQFDPWGQGTLVTVSS SGN-B7H4V light chain variable region 92 EIVMTQSPATLSVSPGERATLSCRASQSVSSNLAWYQQKPGQAPRLLIYGASTRATGIPARFSGSGSGTEFTLTISSLQSEDFAVYYCQQYHSFPFTFGGGTKVEIK SGN-B7H4V VH CDR1 93 GSIKSGSYYWG SGN-B7H4V VH CDR2 94 NIYYSGSTYYNPSLRS SGN-B7H4V VH CDR3 95 AREGSYPNQFDP SGN-B7H4V VL CDR1 96 RASQSVSSNLA SGN-B7H4V VL CDR2 97 GASTRAT SGN-B7H4V VL CDR3 98 QQYHSFPFT dicituzumab vedotin heavy chain 99 EVQLVQSGAEVKKPGATVKISCKVSGYTFTDYYIHWVQQAPGKGLEWMGRVNPDHGDSYYNQKFKDKATITADKSTDTAYMELSSLRSEDTAVYFCARNYLFDHWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK dicituzumab vedotin light chain 100 DIQMTQSPSSVSASVGDRVTITCKASQDVGTAVAWYQQKPGKAPKLLIYWASIRHTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCHQFATYTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Rivatuzumab vedotin heavy chain 101 EVQLVESGGGLVQPGGSLRLSCAASGFSFSDFAMSWVRQAPGKGLEWVATIGRVAFHTYYPDSMKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARHRGFDVGHFDFWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Rivatuzumab vedotin light chain 102 DIQMTQSPSSLSASVGDRVTITCRSSETLVHSSGNTYLEWYQQKPGKAPKLLIYRVSNRFSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCFQGSFNPLTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Ennozumab vedotin (EV) heavy chain variable region 103 EVQLVESGGGLVQPGGSLRLSCAASGFTFSSYNMNWVRQAPGKGLEWVSYISSSSSTIYYADSVKGRFTISRDNAKNSLSLQMNSLRDEDTAVYYCARAYYYGMDVWGQGTTVTVSS Ennozumab vedotin (EV) light chain variable region 104 DIQMTQSPSSVSASVGDRVTITCRASQGISGWLAWYQQKPGKAPKFLIYAASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANSFPPTFGGGTKVEIK Ennozumab vedotin (EV) VH CDR1 105 SYNMN Ennozumab vedotin (EV) VH CDR2 106 YISSSSSTIYYADSVKG Ennozumab vedotin (EV) VH CDR3 107 AYYYGMDV Ennozumab vedotin (EV) VL CDR1 108 RASQGISGWLA Ennozumab vedotin (EV) VL CDR2 109 AASTLQS Ennozumab vedotin (EV) VL CDR3 110 QQANSFPPT h2A2 heavy chain variable region 111 QFQLVQSGAEVKKPGASVKVSCKASGYSFTDYNVNWVRQAPGQGLEWIGVINPKYGTTRYNQKFKGRATLTVDKSTSTAYMELSSLRSEDTAVYYCTRGLNAWDYWGQGTLVTVSS h2A2 light chain variable region 112 DIQMTQSPSSLSASVGDRVTITCGASENIYGALNWYQQKPGKAPKLLIYGATNLEDGVPSRFSGSGSGRDYTFTISSLQPEDIATYYCQNVLTTPYTFGQGTKLEIK h2A2 VH CDR1 113 DYNVN h2A2 VH CDR2 114 VINPKYGTTRYNQKFKG h2A2 VH CDR3 115 GLNAWDY h2A2 VL CDR1 116 GASENIYGALN h2A2 VL CDR2 117 GATNLED h2A2 VL CDR3 118 QNVLTTPYT h15H3 heavy chain variable region 119 QVQLVQSGAEVKKPGASVKVSCKASGYSFSGYFMNWVRQAPGQGLEWMGLINPYNGDSFYNQKFKGRVTMTRQTSTSTVYMELSSLRSEDTAVYYCVRGLRRDFDYWGQGTLVTVSS h15H3 light chain variable region 120 DVVMTQSPLSLPVTLGQPASISCKSSQSLLDSDGKTYLNWLFQRPGQSPRRLIYLVSELDSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCWQGTHFPRTFGGGTKLEIK h15H3 VH CDR1 121 GYFMN h15H3 VH CDR2 122 LINPYNGDSFYNQKFKG h15H3 VH CDR3 123 GLRRDFDY h15H3 VL CDR1 124 KSSQSLLDSDGKTYLN h15H3 VL CDR2 125 LVSELDS h15H3 VL CDR3 126 WQGTHFPRT SGN-CD228A heavy chain variable region 127 QVQLQESGPGLVKPSETLSLTCTVSGDSITSGYWNWIRQPPGKGLEYIGYISDSGITYYNPSLKSRVTISRDTSKNQYSLKLSSVTAADTAVYYCARRTLATYYAMDYWGQGTLVTVSS SGN-CD228A light chain variable region 128 DFVMTQSPLSLPVTLGQPASISCRASQSLVHSDGNTYLHWYQQRPGQSPRLLIYRVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCSQSTHVPPTFGQGTKLEIK SGN-CD228A VH CDR1 129 SGYWN SGN-CD228A VH CDR2 130 YISDSGITYYNPSLKS SGN-CD228A VH CDR3 131 RTLATYYAMDY SGN-CD228A VL CDR1 132 RASQSLVHSDGNTYLH SGN-CD228A VL CDR2 133 RVSNRFS SGN-CD228A VL CDR3 134 SQSTHVPPT SGN-LIV1A latstuzumab vedotin (LV) heavy chain variable region 135 QVQLVQSGAEVKKPGASVKVSCKASGLTIEDYYMHWVRQAPGQGLEWMGWIDPENGDTEYGPKFQGRVTMTRDTSINTAYMELSRLRSDDTAVYYCAVHNAHYGTWFAYWGQGTLVTVSS SGN-LIV1A Ladetuzumab vedotin (LV) light chain variable region 136 DVVMTQSPLSLPVTLGQPASISCRSSQSLLHSSGNTYLEWYQQRPGQSPRPLIYKISTRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYYCFQGSHVPYTFGGGTKVEIK SGN-LIV1A Ladetuzumab Vidotine (LV) VH CDR1 137 DYYMH SGN-LIV1A Ladetuzumab vedotin (LV) VH CDR2 138 WIDPENGDTEYGPKFQG SGN-LIV1A Ladetuzumab vedotin (LV) VH CDR3 139 HNAHYGTWFAY SGN-LIV1A Ladetuzumab Vidotine (LV) VL CDR1 140 RSSQSLLHSSGNTYLE SGN-LIV1A Ladetuzumab vedotin (LV) VL CDR2 141 KISTRFS SGN-LIV1A Ladetuzumab Vidotine (LV) VL CDR3 142 FQGSHVPYT tesotuzumab vedotin (TV) heavy chain variable region 143 EVQLLESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGDYTYYTDSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARSPWGYYLDSWGQGTLVTVSS tesotuzumab vedotin (TV) light chain variable region 144 DIQMTQSPPSLSASAGDRVTITCRASQGISSRLAWYQQKPEKAPKSLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYNSYPYTFGQGTKLEIK tesotuzumab vedotin (TV) VH CDR1 145 GFTFSNYA tesotuzumab vedotin (TV) VH CDR2 146 ISGSGDYT tesotuzumab vedotin (TV) VH CDR3 147 ARSPWGYYLDS Tesotumumab vedotin (TV) VL CDR1 148 QGISSR Tesotumumab vedotin (TV) VL CDR2 149 AAS Tesotumumab vedotin (TV) VL CDR3 150 QQYNSYPYT

圖 1展示非定向化學治療劑削弱T細胞反應。 Figure 1 shows that non-targeted chemotherapeutics impair T cell responses.

圖 2展示CD30+CD8 T細胞之本妥昔單抗維多汀(BV)治療。已經定向遞送至T細胞之維多汀ADC不會抑制增殖。 Figure 2 shows Bentuximab Verdotin (BV) treatment of CD30+CD8 T cells. Vidotin ADCs that have been targeted to T cells do not inhibit proliferation.

圖 3A 至圖 3E展示與具有不同有效負載之ADC相比，內質網(ER)應激誘導對奧瑞他汀抗體-藥物結合物(ADC)，諸如基於維多汀之ADC係優異的。圖3A展示各種臨床上批准之ADC有效負載之表。圖3B展示ER應激傳信反應之圖。圖3C展示用在IC50濃度下具有不同有效負載之ADC或太平洋紫杉醇處理MIA-PaCa-2細胞36或48小時的西方墨點分析。圖3D至圖3E展示用劑量範圍內(圖3D)或IC50劑量(細胞毒性) (圖3E)下具有不同有效負載之ADC處理的表現CHOP驅動螢光素酶報導子(Signosis, Inc.)之MIA-PaCa-2細胞。與未經處理細胞相比，CHOP誘導藉由倍數誘導表現。 Figures 3A - 3E show that induction of endoplasmic reticulum (ER) stress is superior to auristatin antibody-drug conjugates (ADCs), such as vedottin-based ADCs, compared to ADCs with different payloads. Figure 3A shows a table of various clinically approved ADC payloads. Figure 3B shows a graph of ER stress signaling responses. Figure 3C shows Western blot analysis of MIA-PaCa-2 cells treated with ADCs with different payloads at IC50 concentrations or paclitaxel for 36 or 48 hours. Figures 3D-3E show the performance of the CHOP-driven luciferase reporter (Signosis, Inc.) treated with ADCs with different payloads over a range of doses (Figure 3D) or at IC50 doses (cytotoxicity) (Figure 3E). MIA-PaCa-2 cells. CHOP induction was manifested by fold induction compared to untreated cells.

圖 4A 至圖 4B展示臨床ADC有效負載之免疫原性細胞死亡(ICD)潛力。自用具有不同有效負載之1 µg/mL ADC處理72小時之MIA-PaCa-2胰臟腫瘤細胞收集上清液。圖4A展示如藉由Cell Titer Glo測定之TP釋放。圖4B展示如藉由ELISA測定之HMGB1分泌。 Figures 4A - 4B show the immunogenic cell death (ICD) potential of clinical ADC payloads. Supernatants were collected from MIA-PaCa-2 pancreatic tumor cells treated with 1 μg/mL ADC with different payloads for 72 hours. Figure 4A shows TP release as determined by Cell Titer Glo. Figure 4B shows HMGB1 secretion as determined by ELISA.

圖 5A 至圖 5C展示ADC有效負載之免疫活化評估。在周邊血液單核細胞(PBMC)與給藥具有不同有效負載之ADC之L540cy細胞的48小時共培育(在IC50濃度下24小時)之後，藉由流式細胞量測術評估PBMC內之骨髓細胞上的MHC II類(HLA-DR)之上調。藉由針對細胞介素含量之Luminex多工分析評估24小時上清液。圖5A展示藉由ADC之免疫活化。圖5B展示回應於ADC暴露之單核球上之MHCII表現。圖5C展示作為免疫細胞活性之量測的回應於ADC暴露之先天細胞介素CXCL-10/IP10表現。 Figures 5A - 5C show immune activation assessments of ADC payloads. Myeloid cells within PBMCs were assessed by flow cytometry after 48 hours of co-incubation of peripheral blood mononuclear cells (PBMCs) with L540cy cells administered with ADCs with different payloads (24 hours at IC50 concentrations) Up-regulation of MHC class II (HLA-DR) on . Twenty-four hour supernatants were assessed by Luminex multiplex assay for interleukin content. Figure 5A shows immune activation by ADCs. Figure 5B shows MHCII performance on monocytes in response to ADC exposure. Figure 5C shows the expression of the innate interleukin CXCL-10/IP10 in response to ADC exposure as a measure of immune cell activity.

圖 6A 至圖 6E展示關於曲妥珠單抗主鏈之有效負載評估。圖6A展示經評估之曲妥珠單抗ADC的表。圖6B展示ER應激傳信反應之圖。圖6C展示用ADC或藥物處理72小時之BT474細胞的西方墨點。圖6D至圖6E展示維多汀ADC展現多個ICD標誌之強活化。以1 µg/mL給藥之曲妥珠單抗ADC或游離MMAE (100 nM)展現表現SKBR3 HER2之乳癌細胞的不同ICD反應。48或72小時處理之後，收集培養基且用於量測ATP釋放(圖6D)及HMGB1含量(圖6E)。 Figures 6A - 6E show payload assessments for the trastuzumab backbone. Figure 6A shows a table of evaluated trastuzumab ADCs. Figure 6B shows a graph of ER stress signaling responses. Figure 6C shows Western blots of BT474 cells treated with ADC or drugs for 72 hours. Figures 6D-6E show that vedotin ADCs exhibit strong activation of multiple ICD markers. Trastuzumab ADC or free MMAE (100 nM) dosed at 1 µg/mL exhibited differential ICD responses of breast cancer cells expressing SKBR3 HER2. After 48 or 72 hours of treatment, the medium was collected and used to measure ATP release (FIG. 6D) and HMGB1 content (FIG. 6E).

圖 7A進一步說明免疫原性細胞死亡(ICD)路徑。 Figure 7A further illustrates the immunogenic cell death (ICD) pathway.

圖 7B提供關於圖7B至圖7E中使用之某些ADC之有效負載的資訊。 Figure 7B provides information on the payload of some of the ADCs used in Figures 7B-7E.

圖 7C 至圖 7F展示與美登素-ADC (圖7C)、喜樹鹼-ADC (圖7D)、蒽環黴素-ADC (圖7E)及卡奇黴素-ADC (圖7F)相比，回應於用MMAE-ADC進行之處理而產生的JNK傳信活化。 Figures 7C - 7F show comparison to maytansine-ADC (Figure 7C), camptothecin-ADC (Figure 7D), anthracycline-ADC (Figure 7E) and calicheamicin-ADC (Figure 7F) , activation of JNK signaling in response to treatment with MMAE-ADC.

圖 8A 至圖 8D展示與美登素-ADC (圖8A)、喜樹鹼-ADC (圖8B)、蒽環黴素-ADC (圖8C)及其他類型的ADC，包括奧佐黴素(ozogamycin)-ADC、特絲胺酸(teserine)-ADC及AT-ADC (圖8D)相比，回應於用MMAE-ADC進行之處理而產生的CHOP誘導。 Figures 8A - 8D show interactions with maytansine-ADC (Figure 8A), camptothecin-ADC (Figure 8B), anthracycline-ADC (Figure 8C), and other types of ADCs, including ozogamycin )-ADC, teserine-ADC and AT-ADC (FIG. 8D), CHOP induction in response to treatment with MMAE-ADC.

圖 9A 至圖 9D展示與美登素-ADC (圖9A)、喜樹鹼-ADC (圖9B)、蒽環黴素-ADC (圖9C)及其他類型的ADC，包括奧佐黴素-ADC、特絲胺酸-ADC及AT-ADC (圖9D)相比，回應於用MMAE-ADC進行之處理而產生的ATP及HMGB1之釋放。 Figures 9A - 9D show interactions with maytansine-ADC (Figure 9A), camptothecin-ADC (Figure 9B), anthracycline-ADC (Figure 9C), and other types of ADCs, including ozocin-ADC The release of ATP and HMGB1 in response to treatment with MMAE-ADC was compared with , Teserine-ADC and AT-ADC (FIG. 9D).

圖 10A 至圖 10D展示與美登素-ADC (圖10A)、喜樹鹼-ADC (圖10B)、蒽環黴素-ADC (圖10C)及其他類型的ADC，包括奧佐黴素-ADC及特絲胺酸-ADC (圖10D)相比，回應於用MMAE-ADC進行之處理而產生的MHCII表現及CXCL-10/IP10釋放。 Figures 10A - 10D show interactions with maytansine-ADC (Figure 10A), camptothecin-ADC (Figure 10B), anthracycline-ADC (Figure 10C), and other types of ADCs, including ozocin-ADC MHCII expression and CXCL-10/IP10 release in response to treatment with MMAE-ADC compared to Teserine-ADC (FIG. 10D).

圖 10E概括圖7B至圖7E、圖8A至圖8D、圖9A至圖9D及圖10A至圖10D中之結果。 10E summarizes the results in FIGS . 7B-7E, 8A-8D, 9A-9D, and 10A-10D.

圖 11A 至圖 11D展示結合於中國倉鼠卵巢(CHO細胞)中之FcγRIIa、FcγRIIb及FcγRIIIa之抗體。圖11A展示評估之抗體。圖11B展示結合於FcγRIIa。圖11C展示結合於FcγRIIIa。圖11D展示結合於FcγRIIb。 Figures 11A - 11D show antibodies that bind to FcyRIIa, FcyRIIb and FcyRIIIa in Chinese Hamster Ovary (CHO cells). Figure 11A shows the antibodies evaluated. Figure 11B shows binding to FcyRIIa. Figure 11C shows binding to FcyRIIIa. Figure 1 ID shows binding to FcyRIIb.

圖 12A 至圖 12D展示用CD40促效劑及化學療法處理之MIA-PaCa 2胰臟癌細胞中之CXCL10含量(圖12A)、IFNγ含量(圖12B)、IL10含量(圖12C)及巨噬細胞衍生之細胞介素含量(MDC；CCL22) (圖12D)。 12A - 12D show CXCL10 content ( FIG . 12A), IFNγ content (FIG. 12B), IL10 content (FIG. 12C) and macrophages in MIA-PaCa2 pancreatic cancer cells treated with CD40 agonist and chemotherapy Derived interleukin content (MDC; CCL22) (FIG. 12D).

圖 13A 至圖 13D展示用CD40促效劑及化學療法處理之黑素瘤細胞株中之CXCL10含量(圖13A至圖13B)及IL10含量(圖13C至圖13D)。 Figures 13A - 13D show CXCL10 content (Figures 13A-13B) and IL10 content (Figures 13C-13D) in melanoma cell lines treated with CD40 agonists and chemotherapy.

圖 14A 至圖 14C展示來自黑素瘤、肺、乳房及胰臟之腫瘤細胞中之CXCL14含量(圖14A)、IL14含量(圖14B)及IFNγ含量(圖14C)。 Figures 14A - 14C show CXCL14 content (Figure 14A), IL14 content (Figure 14B) and IFNy content (Figure 14C) in tumor cells from melanoma, lung, breast and pancreas.

圖 15展示如藉由腫瘤目標抗原為Thy1.1之人類CD40轉殖基因模型評估的SEA-CD40維多汀與ADC化學療法組合的組合之活體內資料。 Figure 15 shows in vivo data for the combination of SEA-CD40 vedotin and ADC chemotherapy as assessed by a human CD40 transgenic model with the tumor target antigen Thyl.1.

圖 16展示用針對腫瘤相關抗原之ADC-MMAE與具有各種效應功能主鏈之TIGIT靶向抗體的組合處理之腫瘤細胞株中之CXCL10含量。 Figure 16 shows CXCL10 content in tumor cell lines treated with a combination of ADC-MMAE against tumor-associated antigens and TIGIT-targeting antibodies with various effector function backbones.

圖 17展示用針對腫瘤相關抗原之ADC-MMAE與具有各種效應功能主鏈之TIGIT靶向抗體的組合處理之腫瘤細胞株中之IFNγ含量。 Figure 17 shows IFNy levels in tumor cell lines treated with a combination of ADC-MMAE against tumor-associated antigens and TIGIT-targeting antibodies with various effector function backbones.

圖 18A 至圖 18C展示表明未經岩藻糖基化TIGIT抗體及vc-MMAE ADC (「維多汀ADC」)之增強活性的活體外及活體內資料。圖18A展示當此等抗體與經靶向vc-MMAE ADC殺死之腫瘤細胞共培養時，與具有效應子剔除式主鏈(LALA)或標準野生型IgG1 Fc主鏈之對應TIGIT抗體相比，具有增強型(未經岩藻糖基化) IgG1 Fc主鏈之未經岩藻糖基化TIGIT抗體(SEA-TGT)在經由細胞介素IP10誘導驅動免疫活化時顯著更佳。與SEA-TGT之組合展現協同性免疫細胞活化。圖18B及圖18C展示在移植有CT26同基因型腫瘤細胞(圖18B)或Renca同基因型腫瘤細胞(圖18C)之小鼠用以下處理時之抗腫瘤反應：1)次佳劑量之ADC (圖18B中之Thy1.1 vc-MMAE ADC及圖18C中之EphA2 vc-MMAE ADC)；2)一系列次佳劑量之mIgG2a SEA-TGT (經重新格式化呈對應於未經岩藻糖基化人類IgG1主鏈之未經岩藻糖基化小鼠IgG2a形式的SEA-TGT抗體)；或3)兩種藥劑之組合。如自此等圖式可見，共投與此等兩種藥劑顯著增大處理功效，包括在此等不同腫瘤模型中產生高百分比治癒性反應。 Figures 18A - 18C show in vitro and in vivo data demonstrating the enhanced activity of unfucosylated TIGIT antibodies and vc-MMAE ADCs ("vedottin ADCs"). Figure 18A shows that when these antibodies were co-cultured with tumor cells killed by targeting vc-MMAE ADC, compared to the corresponding TIGIT antibodies with effector knockout backbone (LALA) or standard wild-type IgGl Fc backbone, The non-fucosylated TIGIT antibody (SEA-TGT) with an enhanced (non-fucosylated) IgGl Fc backbone was significantly better at driving immune activation via interferon IP10 induction. The combination with SEA-TGT exhibited synergistic immune cell activation. Figures 18B and 18C show the antitumor response in mice engrafted with CT26 isogenic tumor cells (Figure 18B) or Renca isogenic tumor cells (Figure 18C) when treated with: 1) a suboptimal dose of ADC ( Thy1.1 vc-MMAE ADC in Figure 18B and EphA2 vc-MMAE ADC in Figure 18C); 2) a series of suboptimal doses of mIgG2a SEA-TGT (reformatted to correspond to unfucosylated SEA-TGT antibody in the unfucosylated mouse IgG2a form of the human IgGl backbone); or 3) a combination of the two agents. As can be seen from these figures, co-administration of these two agents significantly increased treatment efficacy, including producing a high percentage of curative responses in these different tumor models.

圖 19展示表明未經岩藻糖基化TIGIT抗體(SEA-TGT)及SGN-B7H4維多汀ADC (B7H4V)之增強活性的活體內資料。圖19展示當移植有Renca同基因型腫瘤細胞之小鼠用次治療性劑量之SEA-TGT及次治療性劑量之B7H4V或次治療性劑量之SEA-TGT及治療性劑量之奧沙利鉑(oxaliplatin)處理時之抗腫瘤反應。如圖19中所展示，共投與SEA-TGT及B7H4V顯著增大處理功效，即使係以次治療性劑量，包括產生高百分比治癒反應。 Figure 19 shows in vivo data demonstrating enhanced activity of unfucosylated TIGIT antibody (SEA-TGT) and SGN-B7H4 vedotin ADC (B7H4V). Figure 19 shows when mice engrafted with Renca isogenic tumor cells were treated with a subtherapeutic dose of SEA-TGT and a subtherapeutic dose of B7H4V or a subtherapeutic dose of SEA-TGT and a therapeutic dose of oxaliplatin ( antitumor response to oxaliplatin treatment. As shown in Figure 19, co-administration of SEA-TGT and B7H4V significantly increased treatment efficacy, even at sub-therapeutic doses, including producing a high percent cure response.

圖 20A 至圖 20B展示SEA-CD70 (未經岩藻糖基化抗CD70抗體)及SGN-35 (含有MMAE之抗CD30 ADC)的組合效果。 圖 20A展示非霍奇金氏淋巴瘤(NHL)異種移植模型之活體內腫瘤生長評估。 圖 20B展示NHL異種移植模型在500 mm 3腫瘤大小終點時之Kaplan-Meyer存活評估。Tx (處理)及箭頭指示處理起始日(移植後第19天)。 Figures 20A - 20B show the combined effect of SEA-CD70 (non-fucosylated anti-CD70 antibody) and SGN-35 (anti-CD30 ADC containing MMAE). Figure 20A shows in vivo tumor growth assessment in a non-Hodgkin's lymphoma (NHL) xenograft model. Figure 20B shows the Kaplan-Meyer survival assessment in the NHL xenograft model at the 500 mm3 tumor size endpoint. Tx (treatment) and arrows indicate the day of treatment initiation (day 19 post-implantation).

圖 21A 至圖 21B展示SEA-BCMA (未經岩藻糖基化抗BCMA抗體)及SGN-CD48A (含有MMAE之抗CD48 ADC)之協同效果。圖21A展示異種移植模型之活體內存活評估，且圖21B展示異種移植模型之活體內螢光素酶評估。 Figures 21A - 21B show the synergistic effect of SEA-BCMA (non-fucosylated anti-BCMA antibody) and SGN-CD48A (anti-CD48 ADC containing MMAE). Figure 21A shows the in vivo survival assessment of the xenograft model, and Figure 21B shows the in vivo luciferase assessment of the xenograft model.

圖 22A 至圖 22C展示維多汀ADC誘發活體內免疫細胞募集及活化。圖22A：自動物分離之腫瘤異種移植用vc-MMAE ADC或未結合vc-MMAE同種型ADC處理8天且經受流式細胞量測術或細胞介素分析。圖22B：CD45陽性免疫細胞經CD11c染色且藉由染色細胞表面上之MHC II類表現而觀測到活化。圖22C：藉由Luminex量測瘤內細胞介素。 Figures 22A - 22C show that vedotin ADCs induce in vivo immune cell recruitment and activation. Figure 22A: Tumor xenografts isolated from animals were treated with vc-MMAE ADC or unconjugated vc-MMAE isotype ADC for 8 days and subjected to flow cytometry or interleukin analysis. Figure 22B: CD45 positive immune cells were stained with CD11c and activation was observed by MHC class II expression on the stained cell surface. Figure 22C: Measurement of intratumoral interleukins by Luminex.

圖 23A 至圖 23B展示藉由vc-MMAE ADC誘導T細胞記憶。在Renca同基因型模型中，小鼠藉由vc-MMAE ADC處理治癒(圖23A)。藉由該處理治癒之小鼠經Renca腫瘤細胞再攻擊且彼等小鼠會排斥隨後移植之腫瘤細胞(圖23B)。 Figures 23A - 23B show induction of T cell memory by vc-MMAE ADC. In the Renca isogenic model, mice were cured by vc-MMAE ADC treatment (Figure 23A). Mice cured by this treatment were rechallenged with Renca tumor cells and they rejected the subsequently transplanted tumor cells (FIG. 23B).

圖 24展示由經MMAE或vc-MMAE ADC處理之細胞賦予的保護性抗腫瘤免疫。A20癌細胞用本妥昔單抗維多汀(BV)或MMAE處理且向小鼠投與正在死亡及死亡的細胞。圖24展示免疫接種小鼠呈現較強免疫反應，排斥隨後移植之A20細胞。 Figure 24 shows protective antitumor immunity conferred by cells treated with MMAE or vc-MMAE ADC. A20 cancer cells were treated with Bentuximab vedotin (BV) or MMAE and mice were administered dying and dying cells. Figure 24 shows that the immunized mice exhibited a stronger immune response, rejecting the subsequently transplanted A20 cells.

圖 25展示藉由未經岩藻糖基化抗CD40抗體SEA-CD40之受體聚集及促效作用的例示性模型；及藉由未經岩藻糖基化抗TIGIT抗體SEA-TGT之受體促效作用及突觸形成的例示性模型。如所展示，SEA-CD40抗體可結合表現於抗原呈遞細胞(APC)上之CD-40，其中抗體之Fc部分結合於表現於自然殺手(NK)細胞或單核球上之FcγRIIIa，其促進受體聚集。相比之下，SEA-TGT抗體結合於表現於T細胞上之TIGIT且抗體之Fc區結合於表現於APC上之FcγRIIIa。 Figure 25 shows an exemplary model of receptor aggregation and agonism by non-fucosylated anti-CD40 antibody SEA-CD40; and receptor by non-fucosylated anti-TIGIT antibody SEA-TGT Exemplary models of agonism and synapse formation. As shown, the SEA-CD40 antibody can bind to CD-40 expressed on antigen presenting cells (APCs), wherein the Fc portion of the antibody binds to FcγRIIIa expressed on natural killer (NK) cells or monocytes, which promotes receptor body aggregation. In contrast, the SEA-TGT antibody binds to TIGIT expressed on T cells and the Fc region of the antibody binds to FcyRIIIa expressed on APCs. 

         
          <![CDATA[<110>  美商西健公司(SEAGEN INC.)]]>
          <![CDATA[<120>  組合療法]]>
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          <![CDATA[<150>  US 63/111,045]]>
          <![CDATA[<151>  2020-11-08]]>
          <![CDATA[<150>  US 63/172,411]]>
          <![CDATA[<151>  2021-04-08]]>
          <![CDATA[<150>  US 63/208,179]]>
          <![CDATA[<151>  2021-06-08]]>
          <![CDATA[<160>  150   ]]>
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          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13B及13D VH CDR2]]>
          <![CDATA[<400>  12]]>
          Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe Gln 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  13]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13C VH CDR2]]>
          <![CDATA[<400>  13]]>
          Ser Ile Ile Pro Leu Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe Gln 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  14]]>
          <![CDATA[<211>  18]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13及13A VH CDR3]]>
          <![CDATA[<400>  14]]>
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe 
          1               5                   10                  15      
          Asp Pro 
          <![CDATA[<210>  15]]>
          <![CDATA[<211>  18]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13B VH CDR3]]>
          <![CDATA[<400>  15]]>
          Ala Arg Gly Pro Ser Glu Val Ser Gly Ile Leu Gly Tyr Val Trp Phe 
          1               5                   10                  15      
          Asp Pro 
          <![CDATA[<210>  16]]>
          <![CDATA[<211>  18]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13C及13D VH CDR3]]>
          <![CDATA[<400>  16]]>
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe 
          1               5                   10                  15      
          Asp Pro 
          <![CDATA[<210>  17]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13、13A、13B、13C及13D VL CDR1]]>
          <![CDATA[<400>  17]]>
          Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp 
          1               5                   10                  15      
          <![CDATA[<210>  18]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13、13A、13B、13C及13D VL CDR2]]>
          <![CDATA[<400>  18]]>
          Leu Gly Ser Asn Arg Ala Ser 
          1               5           
          <![CDATA[<210>  19]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13、13A、13B、13C及13D VL CDR3]]>
          <![CDATA[<400>  19]]>
          Met Gln Ala Arg Arg Ile Pro Ile Thr 
          1               5                   
          <![CDATA[<210>  20]]>
          <![CDATA[<211>  455]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13重鏈hIgG1 (及非岩藻糖基化hIgG1)胺基酸序列]]>
          <![CDATA[<400>  20]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr 
                      20                  25                  30          
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Ser Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe 
                      100                 105                 110         
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 
                  115                 120                 125             
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 
              130                 135                 140                 
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 
          145                 150                 155                 160 
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 
                          165                 170                 175     
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 
                      180                 185                 190         
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 
                  195                 200                 205             
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 
              210                 215                 220                 
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 
          225                 230                 235                 240 
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 
                          245                 250                 255     
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 
                      260                 265                 270         
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 
                  275                 280                 285             
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 
              290                 295                 300                 
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 
          305                 310                 315                 320 
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 
                          325                 330                 335     
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 
                      340                 345                 350         
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 
                  355                 360                 365             
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 
              370                 375                 380                 
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 
          385                 390                 395                 400 
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 
                          405                 410                 415     
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 
                      420                 425                 430         
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 
                  435                 440                 445             
          Leu Ser Leu Ser Pro Gly Lys 
              450                 455 
          <![CDATA[<210>  21]]>
          <![CDATA[<211>  455]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13A重鏈hIgG1 (及非岩藻糖基化hIgG1)胺基酸序列]]>
          <![CDATA[<400>  21]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser 
                      20                  25                  30          
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Ser Leu Ile Pro Tyr Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe 
                      100                 105                 110         
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 
                  115                 120                 125             
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 
              130                 135                 140                 
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 
          145                 150                 155                 160 
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 
                          165                 170                 175     
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 
                      180                 185                 190         
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 
                  195                 200                 205             
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 
              210                 215                 220                 
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 
          225                 230                 235                 240 
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 
                          245                 250                 255     
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 
                      260                 265                 270         
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 
                  275                 280                 285             
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 
              290                 295                 300                 
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 
          305                 310                 315                 320 
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 
                          325                 330                 335     
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 
                      340                 345                 350         
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 
                  355                 360                 365             
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 
              370                 375                 380                 
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 
          385                 390                 395                 400 
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 
                          405                 410                 415     
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 
                      420                 425                 430         
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 
                  435                 440                 445             
          Leu Ser Leu Ser Pro Gly Lys 
              450                 455 
          <![CDATA[<210>  22]]>
          <![CDATA[<211>  455]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13B重鏈hIgG1 (及非岩藻糖基化hIgG1)胺基酸序列]]>
          <![CDATA[<400>  22]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ala Trp 
                      20                  25                  30          
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Gly Pro Ser Glu Val Ser Gly Ile Leu Gly Tyr Val Trp Phe 
                      100                 105                 110         
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 
                  115                 120                 125             
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 
              130                 135                 140                 
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 
          145                 150                 155                 160 
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 
                          165                 170                 175     
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 
                      180                 185                 190         
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 
                  195                 200                 205             
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 
              210                 215                 220                 
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 
          225                 230                 235                 240 
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 
                          245                 250                 255     
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 
                      260                 265                 270         
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 
                  275                 280                 285             
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 
              290                 295                 300                 
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 
          305                 310                 315                 320 
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 
                          325                 330                 335     
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 
                      340                 345                 350         
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 
                  355                 360                 365             
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 
              370                 375                 380                 
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 
          385                 390                 395                 400 
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 
                          405                 410                 415     
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 
                      420                 425                 430         
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 
                  435                 440                 445             
          Leu Ser Leu Ser Pro Gly Lys 
              450                 455 
          <![CDATA[<210>  23]]>
          <![CDATA[<211>  455]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13C重鏈hIgG1 (及非岩藻糖基化hIgG1)胺基酸序列]]>
          <![CDATA[<400>  23]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser 
                      20                  25                  30          
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Ser Ile Ile Pro Leu Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe 
                      100                 105                 110         
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 
                  115                 120                 125             
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 
              130                 135                 140                 
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 
          145                 150                 155                 160 
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 
                          165                 170                 175     
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 
                      180                 185                 190         
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 
                  195                 200                 205             
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 
              210                 215                 220                 
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 
          225                 230                 235                 240 
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 
                          245                 250                 255     
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 
                      260                 265                 270         
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 
                  275                 280                 285             
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 
              290                 295                 300                 
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 
          305                 310                 315                 320 
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 
                          325                 330                 335     
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 
                      340                 345                 350         
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 
                  355                 360                 365             
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 
              370                 375                 380                 
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 
          385                 390                 395                 400 
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 
                          405                 410                 415     
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 
                      420                 425                 430         
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 
                  435                 440                 445             
          Leu Ser Leu Ser Pro Gly Lys 
              450                 455 
          <![CDATA[<210>  24]]>
          <![CDATA[<211>  455]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13D重鏈hIgG1 (及非岩藻糖基化hIgG1)胺基酸序列]]>
          <![CDATA[<400>  24]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser 
                      20                  25                  30          
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe 
                      100                 105                 110         
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr 
                  115                 120                 125             
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 
              130                 135                 140                 
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 
          145                 150                 155                 160 
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 
                          165                 170                 175     
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser 
                      180                 185                 190         
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys 
                  195                 200                 205             
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 
              210                 215                 220                 
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro 
          225                 230                 235                 240 
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys 
                          245                 250                 255     
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val 
                      260                 265                 270         
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp 
                  275                 280                 285             
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr 
              290                 295                 300                 
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp 
          305                 310                 315                 320 
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu 
                          325                 330                 335     
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg 
                      340                 345                 350         
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys 
                  355                 360                 365             
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp 
              370                 375                 380                 
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys 
          385                 390                 395                 400 
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser 
                          405                 410                 415     
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser 
                      420                 425                 430         
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser 
                  435                 440                 445             
          Leu Ser Leu Ser Pro Gly Lys 
              450                 455 
          <![CDATA[<210>  25]]>
          <![CDATA[<211>  219]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：純系13、13A、13B、13C及13D輕鏈 hκ (及非岩藻糖基化)]]>
                 胺基酸序列
          <![CDATA[<400>  25]]>
          Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly 
          1               5                   10                  15      
          Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 
                      20                  25                  30          
          Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser 
                  35                  40                  45              
          Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro 
              50                  55                  60                  
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
          65                  70                  75                  80  
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala 
                          85                  90                  95      
          Arg Arg Ile Pro Ile Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 
                      100                 105                 110         
          Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 
                  115                 120                 125             
          Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 
              130                 135                 140                 
          Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 
          145                 150                 155                 160 
          Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 
                          165                 170                 175     
          Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 
                      180                 185                 190         
          Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 
                  195                 200                 205             
          Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 
              210                 215                 
          <![CDATA[<210>  26]]>
          <![CDATA[<211>  444]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 (非岩藻糖基化hS2C6)重鏈]]>
          <![CDATA[<400>  26]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 
          1               5                   10                  15      
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ser Phe Thr Gly Tyr 
                      20                  25                  30          
          Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
                  35                  40                  45              
          Ala Arg Val Ile Pro Asn Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Gly Arg Phe Thr Leu Ser Val Asp Asn Ser Lys Asn Thr Ala Tyr 
          65                  70                  75                  80  
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Glu Gly Ile Tyr Trp Trp Gly Gln Gly Thr Leu Val Thr Val 
                      100                 105                 110         
          Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser 
                  115                 120                 125             
          Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys 
              130                 135                 140                 
          Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu 
          145                 150                 155                 160 
          Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu 
                          165                 170                 175     
          Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr 
                      180                 185                 190         
          Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val 
                  195                 200                 205             
          Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro 
              210                 215                 220                 
          Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe 
          225                 230                 235                 240 
          Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val 
                          245                 250                 255     
          Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe 
                      260                 265                 270         
          Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro 
                  275                 280                 285             
          Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr 
              290                 295                 300                 
          Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val 
          305                 310                 315                 320 
          Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala 
                          325                 330                 335     
          Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg 
                      340                 345                 350         
          Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly 
                  355                 360                 365             
          Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro 
              370                 375                 380                 
          Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser 
          385                 390                 395                 400 
          Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln 
                          405                 410                 415     
          Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His 
                      420                 425                 430         
          Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 
                  435                 440                 
          <![CDATA[<210>  27]]>
          <![CDATA[<211>  219]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 (非岩藻糖基化hS2C6)輕鏈]]>
          <![CDATA[<400>  27]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Ser Leu Val His Ser 
                      20                  25                  30          
          Asn Gly Asn Thr Phe Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala 
                  35                  40                  45              
          Pro Lys Leu Leu Ile Tyr Thr Val Ser Asn Arg Phe Ser Gly Val Pro 
              50                  55                  60                  
          Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 
          65                  70                  75                  80  
          Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Ser Gln Thr 
                          85                  90                  95      
          Thr His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
                      100                 105                 110         
          Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 
                  115                 120                 125             
          Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 
              130                 135                 140                 
          Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 
          145                 150                 155                 160 
          Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 
                          165                 170                 175     
          Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 
                      180                 185                 190         
          Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 
                  195                 200                 205             
          Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 
              210                 215                 
          <![CDATA[<210>  28]]>
          <![CDATA[<211>  114]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40之VH]]>
          <![CDATA[<400>  28]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 
          1               5                   10                  15      
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ser Phe Thr Gly Tyr 
                      20                  25                  30          
          Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
                  35                  40                  45              
          Ala Arg Val Ile Pro Asn Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Gly Arg Phe Thr Leu Ser Val Asp Asn Ser Lys Asn Thr Ala Tyr 
          65                  70                  75                  80  
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Glu Gly Ile Tyr Trp Trp Gly Gln Gly Thr Leu Val Thr Val 
                      100                 105                 110         
          Ser Ser 
          <![CDATA[<210>  29]]>
          <![CDATA[<211>  112]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40之VL]]>
          <![CDATA[<400>  29]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Ser Leu Val His Ser 
                      20                  25                  30          
          Asn Gly Asn Thr Phe Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala 
                  35                  40                  45              
          Pro Lys Leu Leu Ile Tyr Thr Val Ser Asn Arg Phe Ser Gly Val Pro 
              50                  55                  60                  
          Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 
          65                  70                  75                  80  
          Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Ser Gln Thr 
                          85                  90                  95      
          Thr His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
                      100                 105                 110         
          <![CDATA[<210>  30]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 VH CDR1]]>
          <![CDATA[<400>  30]]>
          Gly Tyr Tyr Ile His 
          1               5   
          <![CDATA[<210>  31]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 VH CDR2]]>
          <![CDATA[<400>  31]]>
          Arg Val Ile Pro Asn Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  32]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 VH CDR3]]>
          <![CDATA[<400>  32]]>
          Glu Gly Ile Tyr Trp 
          1               5   
          <![CDATA[<210>  33]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 VL CDR1]]>
          <![CDATA[<400>  33]]>
          Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Phe Leu His 
          1               5                   10                  15      
          <![CDATA[<210>  34]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 VL CDR2]]>
          <![CDATA[<400>  34]]>
          Thr Val Ser Asn Arg Phe Ser 
          1               5           
          <![CDATA[<210>  35]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD40 VL CDR3]]>
          <![CDATA[<400>  35]]>
          Ser Gln Thr Thr His Val Pro Trp Thr 
          1               5                   
          <![CDATA[<210>  36]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替代的抗CD40抗體CDR2]]>
          <![CDATA[<400>  36]]>
          Arg Val Ile Pro Gln Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  37]]>
          <![CDATA[<211>  116]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B6A重鏈可變區]]>
          <![CDATA[<400>  37]]>
          Gln Phe Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr 
                      20                  25                  30          
          Asn Val Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
                  35                  40                  45              
          Gly Val Ile Asn Pro Lys Tyr Gly Thr Thr Arg Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Thr Arg Gly Leu Asn Ala Trp Asp Tyr Trp Gly Gln Gly Thr Leu Val 
                      100                 105                 110         
          Thr Val Ser Ser 
                  115     
          <![CDATA[<210>  38]]>
          <![CDATA[<211>  107]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B6A輕鏈可變區]]>
          <![CDATA[<400>  38]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Ala 
                      20                  25                  30          
          Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
                  35                  40                  45              
          Tyr Gly Ala Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Arg Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Asn Val Leu Thr Thr Pro Tyr 
                          85                  90                  95      
          Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
                      100                 105         
          <![CDATA[<210>  39]]>
          <![CDATA[<211>  115]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-STNV重鏈可變區]]>
          <![CDATA[<400>  39]]>
          Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp His 
                      20                  25                  30          
          Ala Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Tyr Phe Ser Pro Gly Asn Asp Asp Ile Lys Tyr Asn Glu Lys Phe 
              50                  55                  60                  
          Arg Gly Arg Val Thr Met Thr Ala Asp Lys Ser Ser Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 
                          85                  90                  95      
          Lys Arg Ser Leu Ser Thr Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr 
                      100                 105                 110         
          Val Ser Ser 
                  115 
          <![CDATA[<210>  40]]>
          <![CDATA[<211>  113]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-STNV重鏈可變區]]>
          <![CDATA[<400>  40]]>
          Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 
          1               5                   10                  15      
          Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Arg 
                      20                  25                  30          
          Gly Asn His Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln 
                  35                  40                  45              
          Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 
              50                  55                  60                  
          Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
          65                  70                  75                  80  
          Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Asn 
                          85                  90                  95      
          Asp Tyr Thr Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile 
                      100                 105                 110         
          Lys 
          <![CDATA[<210>  41]]>
          <![CDATA[<211>  118]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70重鏈可變區]]>
          <![CDATA[<400>  41]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 
                      20                  25                  30          
          Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Lys Trp Met 
                  35                  40                  45              
          Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe 
              50                  55                  60                  
          Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Asp Tyr Gly Asp Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr 
                      100                 105                 110         
          Thr Val Thr Val Ser Ser 
                  115             
          <![CDATA[<210>  42]]>
          <![CDATA[<211>  111]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD70輕鏈可變區]]>
          <![CDATA[<400>  42]]>
          Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly 
          1               5                   10                  15      
          Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Lys Ser Val Ser Thr Ser 
                      20                  25                  30          
          Gly Tyr Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 
                  35                  40                  45              
          Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Asp 
              50                  55                  60                  
          Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser 
          65                  70                  75                  80  
          Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln His Ser Arg 
                          85                  90                  95      
          Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
                      100                 105                 110     
          <![CDATA[<210>  43]]>
          <![CDATA[<211>  119]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A重鏈可變區]]>
          <![CDATA[<400>  43]]>
          Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 
          1               5                   10                  15      
          Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Thr Ser Gly 
                      20                  25                  30          
          Tyr Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Tyr Ile 
                  35                  40                  45              
          Gly Tyr Ile Ser Asp Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys 
              50                  55                  60                  
          Ser Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Ser Leu 
          65                  70                  75                  80  
          Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 
                          85                  90                  95      
          Arg Arg Thr Leu Ala Thr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly 
                      100                 105                 110         
          Thr Leu Val Thr Val Ser Ser 
                  115                 
          <![CDATA[<210>  44]]>
          <![CDATA[<211>  112]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A輕鏈可變區]]>
          <![CDATA[<400>  44]]>
          Asp Phe Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 
          1               5                   10                  15      
          Gln Pro Ala Ser Ile Ser Cys Arg Ala Ser Gln Ser Leu Val His Ser 
                      20                  25                  30          
          Asp Gly Asn Thr Tyr Leu His Trp Tyr Gln Gln Arg Pro Gly Gln Ser 
                  35                  40                  45              
          Pro Arg Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro 
              50                  55                  60                  
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
          65                  70                  75                  80  
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser 
                          85                  90                  95      
          Thr His Val Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
                      100                 105                 110         
          <![CDATA[<210>  45]]>
          <![CDATA[<211>  121]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA重鏈可變區]]>
          <![CDATA[<400>  45]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr 
                      20                  25                  30          
          Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
                  35                  40                  45              
          Gly Tyr Ile Asn Pro Asn Ser Gly Tyr Thr Asn Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Ala Thr Met Thr Ala Asp Lys Ser Ile Asn Thr Ala Tyr 
          65                  70                  75                  80  
          Val Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys 
                          85                  90                  95      
          Thr Arg Tyr Met Trp Glu Arg Val Thr Gly Phe Phe Asp Phe Trp Gly 
                      100                 105                 110         
          Gln Gly Thr Met Val Thr Val Ser Ser 
                  115                 120     
          <![CDATA[<210>  46]]>
          <![CDATA[<211>  107]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA輕鏈可變區]]>
          <![CDATA[<400>  46]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Leu Ala Ser Glu Asp Ile Ser Asp Asp 
                      20                  25                  30          
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Val 
                  35                  40                  45              
          Tyr Thr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Thr Tyr Lys Phe Pro Pro 
                          85                  90                  95      
          Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100                 105             
          <![CDATA[<210>  47]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA VH CDR1]]>
          <![CDATA[<400>  47]]>
          Asp Tyr Tyr Ile His 
          1               5   
          <![CDATA[<210>  48]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA VH CDR2]]>
          <![CDATA[<400>  48]]>
          Tyr Ile Asn Pro Asn Ser Gly Tyr Thr Asn Tyr Ala Gln Lys Phe Gln 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  49]]>
          <![CDATA[<211>  12]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA VH CDR3]]>
          <![CDATA[<400>  49]]>
          Tyr Met Trp Glu Arg Val Thr Gly Phe Phe Asp Phe 
          1               5                   10          
          <![CDATA[<210>  50]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA VL CDR1]]>
          <![CDATA[<400>  50]]>
          Leu Ala Ser Glu Asp Ile Ser Asp Asp Leu Ala 
          1               5                   10      
          <![CDATA[<210>  51]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA VL CDR2]]>
          <![CDATA[<400>  51]]>
          Thr Thr Ser Ser Leu Gln Ser 
          1               5           
          <![CDATA[<210>  52]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-BCMA VL CDR3]]>
          <![CDATA[<400>  52]]>
          Gln Gln Thr Tyr Lys Phe Pro Pro Thr 
          1               5                   
          <![CDATA[<210>  53]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70 VH CDR1]]>
          <![CDATA[<400>  53]]>
          Asn Tyr Gly Met Asn 
          1               5   
          <![CDATA[<210>  54]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70 VH CDR2]]>
          <![CDATA[<400>  54]]>
          Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  55]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70 VH CDR3]]>
          <![CDATA[<400>  55]]>
          Asp Tyr Gly Asp Tyr Gly Met Asp Tyr 
          1               5                   
          <![CDATA[<210>  56]]>
          <![CDATA[<211>  15]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70 VL CDR1]]>
          <![CDATA[<400>  56]]>
          Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met His 
          1               5                   10                  15  
          <![CDATA[<210>  57]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70 VL CDR2]]>
          <![CDATA[<400>  57]]>
          Leu Ala Ser Asn Leu Glu Ser 
          1               5           
          <![CDATA[<210>  58]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SEA-CD70 VL CDR3]]>
          <![CDATA[<400>  58]]>
          Gln His Ser Arg Glu Val Pro Trp Thr 
          1               5                   
          <![CDATA[<210>  59]]>
          <![CDATA[<211>  118]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10)重鏈可變區]]>
          <![CDATA[<400>  59]]>
          Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr 
                      20                  25                  30          
          Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 
                  35                  40                  45              
          Gly Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Thr Arg Ser Trp Arg Gly Asn Ser Phe Asp Tyr Trp Gly Gln Gly Thr 
                      100                 105                 110         
          Thr Leu Thr Val Ser Ser 
                  115             
          <![CDATA[<210>  60]]>
          <![CDATA[<211>  109]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10)輕鏈可變區]]>
          <![CDATA[<400>  60]]>
          Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 
          1               5                   10                  15      
          Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 
                      20                  25                  30          
          Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln 
                  35                  40                  45              
          Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 
              50                  55                  60                  
          Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
          65                  70                  75                  80  
          Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 
                          85                  90                  95      
          Asp Tyr Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys 
                      100                 105                 
          <![CDATA[<210>  61]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10) VH CDR1]]>
          <![CDATA[<400>  61]]>
          Ser Tyr Trp Ile Asn 
          1               5   
          <![CDATA[<210>  62]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10) VH CDR2]]>
          <![CDATA[<400>  62]]>
          Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe Lys 
          1               5                   10                  15      
          Asp 
          <![CDATA[<210>  63]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10) VH CDR3]]>
          <![CDATA[<400>  63]]>
          Ser Trp Arg Gly Asn Ser Phe Asp Tyr 
          1               5                   
          <![CDATA[<210>  64]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10) VL CDR1]]>
          <![CDATA[<400>  64]]>
          Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu 
          1               5                   10                  15      
          Thr 
          <![CDATA[<210>  65]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10) VL CDR2]]>
          <![CDATA[<400>  65]]>
          Trp Ala Ser Thr Arg Glu Ser 
          1               5           
          <![CDATA[<210>  66]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：唑貝妥西單抗(175D10) VL CDR3]]>
          <![CDATA[<400>  66]]>
          Gln Asn Asp Tyr Ser Tyr Pro Phe Thr 
          1               5                   
          <![CDATA[<210>  67]]>
          <![CDATA[<211>  118]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 重鏈可變區]]>
          <![CDATA[<400>  67]]>
          Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 
          1               5                   10                  15      
          Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 
                      20                  25                  30          
          Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met 
                  35                  40                  45              
          Gly Trp Ile Asn Thr Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe 
              50                  55                  60                  
          Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys 
                          85                  90                  95      
          Ala Arg Leu Gly Phe Gly Asn Ala Met Asp Tyr Trp Gly Gln Gly Thr 
                      100                 105                 110         
          Ser Val Thr Val Ser Ser 
                  115             
          <![CDATA[<210>  68]]>
          <![CDATA[<211>  113]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 輕鏈可變區]]>
          <![CDATA[<400>  68]]>
          Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly 
          1               5                   10                  15      
          Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser 
                      20                  25                  30          
          Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln 
                  35                  40                  45              
          Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val 
              50                  55                  60                  
          Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr 
          65                  70                  75                  80  
          Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn 
                          85                  90                  95      
          Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu 
                      100                 105                 110         
          Lys 
          <![CDATA[<210>  69]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 VH CDR1]]>
          <![CDATA[<400>  69]]>
          Asn Tyr Gly Met Asn 
          1               5   
          <![CDATA[<210>  70]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 VH CDR2]]>
          <![CDATA[<400>  70]]>
          Trp Ile Asn Thr Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  71]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 VH CDR3]]>
          <![CDATA[<400>  71]]>
          Leu Gly Phe Gly Asn Ala Met Asp Tyr 
          1               5                   
          <![CDATA[<210>  72]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 VL CDR1]]>
          <![CDATA[<400>  72]]>
          Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu 
          1               5                   10                  15      
          Thr 
          <![CDATA[<210>  73]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 VL CDR2]]>
          <![CDATA[<400>  73]]>
          Trp Ala Ser Thr Arg Glu Ser 
          1               5           
          <![CDATA[<210>  74]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：163E12 VL CDR3]]>
          <![CDATA[<400>  74]]>
          Gln Asn Asp Tyr Ser Tyr Pro Leu Thr 
          1               5                   
          <![CDATA[<210>  75]]>
          <![CDATA[<211>  123]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V重鏈可變區]]>
          <![CDATA[<400>  75]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Thr Ser Gly Asp Thr Phe Ser Thr Ala 
                      20                  25                  30          
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Gly Ile Ile Pro Ile Phe Gly Lys Ala His Tyr Ala Gln Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys 
                          85                  90                  95      
          Ala Arg Lys Phe His Phe Val Ser Gly Ser Pro Phe Gly Met Asp Val 
                      100                 105                 110         
          Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser 
                  115                 120             
          <![CDATA[<210>  76]]>
          <![CDATA[<211>  106]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V輕鏈可變區]]>
          <![CDATA[<400>  76]]>
          Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly 
          1               5                   10                  15      
          Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr 
                      20                  25                  30          
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 
                  35                  40                  45              
          Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro 
          65                  70                  75                  80  
          Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Thr 
                          85                  90                  95      
          Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
                      100                 105     
          <![CDATA[<210>  77]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V VH CDR1]]>
          <![CDATA[<400>  77]]>
          Thr Ala Ala Ile Ser 
          1               5   
          <![CDATA[<210>  78]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V VH CDR2]]>
          <![CDATA[<400>  78]]>
          Gly Ile Ile Pro Ile Phe Gly Lys Ala His Tyr Ala Gln Lys Phe Gln 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  79]]>
          <![CDATA[<211>  14]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V VH CDR3]]>
          <![CDATA[<400>  79]]>
          Lys Phe His Phe Val Ser Gly Ser Pro Phe Gly Met Asp Val 
          1               5                   10                  
          <![CDATA[<210>  80]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V VL CDR1]]>
          <![CDATA[<400>  80]]>
          Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala 
          1               5                   10      
          <![CDATA[<210>  81]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V VL CDR2]]>
          <![CDATA[<400>  81]]>
          Asp Ala Ser Asn Arg Ala Thr 
          1               5           
          <![CDATA[<210>  82]]>
          <![CDATA[<211>  8]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-PDL1V VL CDR3]]>
          <![CDATA[<400>  82]]>
          Gln Gln Arg Ser Asn Trp Pro Thr 
          1               5               
          <![CDATA[<210>  83]]>
          <![CDATA[<211>  123]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV重鏈可變區]]>
          <![CDATA[<400>  83]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg 
          1               5                   10                  15      
          Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Thr Asp Tyr 
                      20                  25                  30          
          Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu 
                  35                  40                  45              
          Ala Leu Ile Arg Asn Lys Ala Thr Gly Tyr Thr Thr Glu Tyr Thr Ala 
              50                  55                  60                  
          Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Ser Ile 
          65                  70                  75                  80  
          Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr 
                          85                  90                  95      
          Tyr Cys Ala Arg Ala Ser Phe Tyr Tyr Asp Gly Lys Val Leu Ala Tyr 
                      100                 105                 110         
          Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser 
                  115                 120             
          <![CDATA[<210>  84]]>
          <![CDATA[<211>  106]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV輕鏈可變區]]>
          <![CDATA[<400>  84]]>
          Asp Thr Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Asn Lys Tyr 
                      20                  25                  30          
          Leu Ala Trp Tyr Gln Tyr Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
                  35                  40                  45              
          His Tyr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Arg Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Asn Leu Tyr Thr 
                          85                  90                  95      
          Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
                      100                 105     
          <![CDATA[<210>  85]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV VH CDR1]]>
          <![CDATA[<400>  85]]>
          Asp Tyr Tyr Met Ser 
          1               5   
          <![CDATA[<210>  86]]>
          <![CDATA[<211>  19]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV VH CDR2]]>
          <![CDATA[<400>  86]]>
          Leu Ile Arg Asn Lys Ala Thr Gly Tyr Thr Thr Glu Tyr Thr Ala Ser 
          1               5                   10                  15      
          Val Lys Gly 
          <![CDATA[<210>  87]]>
          <![CDATA[<211>  12]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV VH CDR3]]>
          <![CDATA[<400>  87]]>
          Ala Ser Phe Tyr Tyr Asp Gly Lys Val Leu Ala Tyr 
          1               5                   10          
          <![CDATA[<210>  88]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV VL CDR1]]>
          <![CDATA[<400>  88]]>
          Gln Ala Ser Gln Asp Ile Asn Lys Tyr Leu Ala 
          1               5                   10      
          <![CDATA[<210>  89]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV VL CDR2]]>
          <![CDATA[<400>  89]]>
          Tyr Thr Ser Ser Leu Gln Ser 
          1               5           
          <![CDATA[<210>  90]]>
          <![CDATA[<211>  8]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-ALPV VL CDR3]]>
          <![CDATA[<400>  90]]>
          Leu Gln Tyr Asp Asn Leu Tyr Thr 
          1               5               
          <![CDATA[<210>  91]]>
          <![CDATA[<211>  120]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V重鏈可變區]]>
          <![CDATA[<400>  91]]>
          Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 
          1               5                   10                  15      
          Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Lys Ser Gly 
                      20                  25                  30          
          Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu 
                  35                  40                  45              
          Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser 
              50                  55                  60                  
          Leu Arg Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe 
          65                  70                  75                  80  
          Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr 
                          85                  90                  95      
          Cys Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro Trp Gly Gln 
                      100                 105                 110         
          Gly Thr Leu Val Thr Val Ser Ser 
                  115                 120 
          <![CDATA[<210>  92]]>
          <![CDATA[<211>  107]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V輕鏈可變區]]>
          <![CDATA[<400>  92]]>
          Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly 
          1               5                   10                  15      
          Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn 
                      20                  25                  30          
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile 
                  35                  40                  45              
          Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser 
          65                  70                  75                  80  
          Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr His Ser Phe Pro Phe 
                          85                  90                  95      
          Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 
                      100                 105         
          <![CDATA[<210>  93]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V VH CDR1]]>
          <![CDATA[<400>  93]]>
          Gly Ser Ile Lys Ser Gly Ser Tyr Tyr Trp Gly 
          1               5                   10      
          <![CDATA[<210>  94]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V VH CDR2]]>
          <![CDATA[<400>  94]]>
          Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Arg Ser 
          1               5                   10                  15      
          <![CDATA[<210>  95]]>
          <![CDATA[<211>  12]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V VH CDR3]]>
          <![CDATA[<400>  95]]>
          Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro 
          1               5                   10          
          <![CDATA[<210>  96]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V VL CDR1]]>
          <![CDATA[<400>  96]]>
          Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala 
          1               5                   10      
          <![CDATA[<210>  97]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V VL CDR2]]>
          <![CDATA[<400>  97]]>
          Gly Ala Ser Thr Arg Ala Thr 
          1               5           
          <![CDATA[<210>  98]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-B7H4V VL CDR3]]>
          <![CDATA[<400>  98]]>
          Gln Gln Tyr His Ser Phe Pro Phe Thr 
          1               5                   
          <![CDATA[<210>  99]]>
          <![CDATA[<211>  445]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：迪西妥單抗維多汀重鏈]]>
          <![CDATA[<400>  99]]>
          Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr 
                      20                  25                  30          
          Tyr Ile His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Arg Val Asn Pro Asp His Gly Asp Ser Tyr Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Asp Lys Ala Thr Ile Thr Ala Asp Lys Ser Thr Asp Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys 
                          85                  90                  95      
          Ala Arg Asn Tyr Leu Phe Asp His Trp Gly Gln Gly Thr Leu Val Thr 
                      100                 105                 110         
          Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro 
                  115                 120                 125             
          Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val 
              130                 135                 140                 
          Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala 
          145                 150                 155                 160 
          Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly 
                          165                 170                 175     
          Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly 
                      180                 185                 190         
          Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys 
                  195                 200                 205             
          Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys 
              210                 215                 220                 
          Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu 
          225                 230                 235                 240 
          Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu 
                          245                 250                 255     
          Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys 
                      260                 265                 270         
          Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys 
                  275                 280                 285             
          Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu 
              290                 295                 300                 
          Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys 
          305                 310                 315                 320 
          Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys 
                          325                 330                 335     
          Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser 
                      340                 345                 350         
          Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys 
                  355                 360                 365             
          Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln 
              370                 375                 380                 
          Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly 
          385                 390                 395                 400 
          Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln 
                          405                 410                 415     
          Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn 
                      420                 425                 430         
          His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys 
                  435                 440                 445 
          <![CDATA[<210>  100]]>
          <![CDATA[<211>  212]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：迪西妥單抗維多汀輕鏈]]>
          <![CDATA[<400>  100]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala 
                      20                  25                  30          
          Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
                  35                  40                  45              
          Tyr Trp Ala Ser Ile Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Phe Ala Thr Tyr Thr Phe 
                          85                  90                  95      
          Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser 
                      100                 105                 110         
          Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala 
                  115                 120                 125             
          Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val 
              130                 135                 140                 
          Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser 
          145                 150                 155                 160 
          Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr 
                          165                 170                 175     
          Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys 
                      180                 185                 190         
          Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn 
                  195                 200                 205             
          Arg Gly Glu Cys 
              210         
          <![CDATA[<210>  101]]>
          <![CDATA[<211>  450]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：立伐土珠單抗維多汀重鏈]]>
          <![CDATA[<400>  101]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 
          1               5                   10                  15      
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Asp Phe 
                      20                  25                  30          
          Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
                  35                  40                  45              
          Ala Thr Ile Gly Arg Val Ala Phe His Thr Tyr Tyr Pro Asp Ser Met 
              50                  55                  60                  
          Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 
          65                  70                  75                  80  
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg His Arg Gly Phe Asp Val Gly His Phe Asp Phe Trp Gly Gln 
                      100                 105                 110         
          Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val 
                  115                 120                 125             
          Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala 
              130                 135                 140                 
          Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser 
          145                 150                 155                 160 
          Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val 
                          165                 170                 175     
          Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro 
                      180                 185                 190         
          Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys 
                  195                 200                 205             
          Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp 
              210                 215                 220                 
          Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly 
          225                 230                 235                 240 
          Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile 
                          245                 250                 255     
          Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu 
                      260                 265                 270         
          Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His 
                  275                 280                 285             
          Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg 
              290                 295                 300                 
          Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys 
          305                 310                 315                 320 
          Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu 
                          325                 330                 335     
          Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr 
                      340                 345                 350         
          Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu 
                  355                 360                 365             
          Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp 
              370                 375                 380                 
          Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val 
          385                 390                 395                 400 
          Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp 
                          405                 410                 415     
          Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His 
                      420                 425                 430         
          Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro 
                  435                 440                 445             
          Gly Lys 
              450 
          <![CDATA[<210>  102]]>
          <![CDATA[<211>  219]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：立伐土珠單抗維多汀輕鏈]]>
          <![CDATA[<400>  102]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Glu Thr Leu Val His Ser 
                      20                  25                  30          
          Ser Gly Asn Thr Tyr Leu Glu Trp Tyr Gln Gln Lys Pro Gly Lys Ala 
                  35                  40                  45              
          Pro Lys Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro 
              50                  55                  60                  
          Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile 
          65                  70                  75                  80  
          Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly 
                          85                  90                  95      
          Ser Phe Asn Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys 
                      100                 105                 110         
          Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu 
                  115                 120                 125             
          Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe 
              130                 135                 140                 
          Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln 
          145                 150                 155                 160 
          Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser 
                          165                 170                 175     
          Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu 
                      180                 185                 190         
          Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser 
                  195                 200                 205             
          Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys 
              210                 215                 
          <![CDATA[<210>  103]]>
          <![CDATA[<211>  117]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV)重鏈可變區]]>
          <![CDATA[<400>  103]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 
          1               5                   10                  15      
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr 
                      20                  25                  30          
          Asn Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
                  35                  40                  45              
          Ser Tyr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val 
              50                  55                  60                  
          Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Ser 
          65                  70                  75                  80  
          Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Ala Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr 
                      100                 105                 110         
          Val Thr Val Ser Ser 
                  115         
          <![CDATA[<210>  104]]>
          <![CDATA[<211>  107]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV)輕鏈可變區]]>
          <![CDATA[<400>  104]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Gly Trp 
                      20                  25                  30          
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile 
                  35                  40                  45              
          Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro 
                          85                  90                  95      
          Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 
                      100                 105         
          <![CDATA[<210>  105]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV) VH CDR1]]>
          <![CDATA[<400>  105]]>
          Ser Tyr Asn Met Asn 
          1               5   
          <![CDATA[<210>  106]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV) VH CDR2]]>
          <![CDATA[<400>  106]]>
          Tyr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  107]]>
          <![CDATA[<211>  8]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV) VH CDR3]]>
          <![CDATA[<400>  107]]>
          Ala Tyr Tyr Tyr Gly Met Asp Val 
          1               5               
          <![CDATA[<210>  108]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV) VL CDR1]]>
          <![CDATA[<400>  108]]>
          Arg Ala Ser Gln Gly Ile Ser Gly Trp Leu Ala 
          1               5                   10      
          <![CDATA[<210>  109]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV) VL CDR2]]>
          <![CDATA[<400>  109]]>
          Ala Ala Ser Thr Leu Gln Ser 
          1               5           
          <![CDATA[<210>  110]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：PADCEV (恩諾單抗維多汀) (EV) VL CDR3]]>
          <![CDATA[<400>  110]]>
          Gln Gln Ala Asn Ser Phe Pro Pro Thr 
          1               5                   
          <![CDATA[<210>  111]]>
          <![CDATA[<211>  116]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2重鏈可變區]]>
          <![CDATA[<400>  111]]>
          Gln Phe Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr 
                      20                  25                  30          
          Asn Val Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile 
                  35                  40                  45              
          Gly Val Ile Asn Pro Lys Tyr Gly Thr Thr Arg Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Thr Arg Gly Leu Asn Ala Trp Asp Tyr Trp Gly Gln Gly Thr Leu Val 
                      100                 105                 110         
          Thr Val Ser Ser 
                  115     
          <![CDATA[<210>  112]]>
          <![CDATA[<211>  107]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2輕鏈可變區]]>
          <![CDATA[<400>  112]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Ala 
                      20                  25                  30          
          Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile 
                  35                  40                  45              
          Tyr Gly Ala Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Arg Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Asn Val Leu Thr Thr Pro Tyr 
                          85                  90                  95      
          Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
                      100                 105         
          <![CDATA[<210>  113]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2 VH CDR1]]>
          <![CDATA[<400>  113]]>
          Asp Tyr Asn Val Asn 
          1               5   
          <![CDATA[<210>  114]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2 VH CDR2]]>
          <![CDATA[<400>  114]]>
          Val Ile Asn Pro Lys Tyr Gly Thr Thr Arg Tyr Asn Gln Lys Phe Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  115]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2 VH CDR3]]>
          <![CDATA[<400>  115]]>
          Gly Leu Asn Ala Trp Asp Tyr 
          1               5           
          <![CDATA[<210>  116]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2 VL CDR1]]>
          <![CDATA[<400>  116]]>
          Gly Ala Ser Glu Asn Ile Tyr Gly Ala Leu Asn 
          1               5                   10      
          <![CDATA[<210>  117]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2 VL CDR2]]>
          <![CDATA[<400>  117]]>
          Gly Ala Thr Asn Leu Glu Asp 
          1               5           
          <![CDATA[<210>  118]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h2A2 VL CDR3]]>
          <![CDATA[<400>  118]]>
          Gln Asn Val Leu Thr Thr Pro Tyr Thr 
          1               5                   
          <![CDATA[<210>  119]]>
          <![CDATA[<211>  117]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>   合成：h15H3重鏈可變區]]>
          <![CDATA[<400>  119]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr 
                      20                  25                  30          
          Phe Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Leu Ile Asn Pro Tyr Asn Gly Asp Ser Phe Tyr Asn Gln Lys Phe 
              50                  55                  60                  
          Lys Gly Arg Val Thr Met Thr Arg Gln Thr Ser Thr Ser Thr Val Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Val Arg Gly Leu Arg Arg Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu 
                      100                 105                 110         
          Val Thr Val Ser Ser 
                  115         
          <![CDATA[<210>  120]]>
          <![CDATA[<211>  112]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3輕鏈可變區]]>
          <![CDATA[<400>  120]]>
          Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 
          1               5                   10                  15      
          Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser 
                      20                  25                  30          
          Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gln Arg Pro Gly Gln Ser 
                  35                  40                  45              
          Pro Arg Arg Leu Ile Tyr Leu Val Ser Glu Leu Asp Ser Gly Val Pro 
              50                  55                  60                  
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
          65                  70                  75                  80  
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Trp Gln Gly 
                          85                  90                  95      
          Thr His Phe Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys 
                      100                 105                 110         
          <![CDATA[<210>  121]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3 VH CDR1]]>
          <![CDATA[<400>  121]]>
          Gly Tyr Phe Met Asn 
          1               5   
          <![CDATA[<210>  122]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3 VH CDR2]]>
          <![CDATA[<400>  122]]>
          Leu Ile Asn Pro Tyr Asn Gly Asp Ser Phe Tyr Asn Gln Lys Phe Lys 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  123]]>
          <![CDATA[<211>  8]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3 VH CDR3]]>
          <![CDATA[<400>  123]]>
          Gly Leu Arg Arg Asp Phe Asp Tyr 
          1               5               
          <![CDATA[<210>  124]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3 VL CDR1]]>
          <![CDATA[<400>  124]]>
          Lys Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn 
          1               5                   10                  15      
          <![CDATA[<210>  125]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3 VL CDR2]]>
          <![CDATA[<400>  125]]>
          Leu Val Ser Glu Leu Asp Ser 
          1               5           
          <![CDATA[<210>  126]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：h15H3 VL CDR3]]>
          <![CDATA[<400>  126]]>
          Trp Gln Gly Thr His Phe Pro Arg Thr 
          1               5                   
          <![CDATA[<210>  127]]>
          <![CDATA[<211>  119]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A重鏈可變區]]>
          <![CDATA[<400>  127]]>
          Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu 
          1               5                   10                  15      
          Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Thr Ser Gly 
                      20                  25                  30          
          Tyr Trp Asn Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu Tyr Ile 
                  35                  40                  45              
          Gly Tyr Ile Ser Asp Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys 
              50                  55                  60                  
          Ser Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Ser Leu 
          65                  70                  75                  80  
          Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala 
                          85                  90                  95      
          Arg Arg Thr Leu Ala Thr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly 
                      100                 105                 110         
          Thr Leu Val Thr Val Ser Ser 
                  115                 
          <![CDATA[<210>  128]]>
          <![CDATA[<211>  112]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A輕鏈可變區]]>
          <![CDATA[<400>  128]]>
          Asp Phe Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 
          1               5                   10                  15      
          Gln Pro Ala Ser Ile Ser Cys Arg Ala Ser Gln Ser Leu Val His Ser 
                      20                  25                  30          
          Asp Gly Asn Thr Tyr Leu His Trp Tyr Gln Gln Arg Pro Gly Gln Ser 
                  35                  40                  45              
          Pro Arg Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro 
              50                  55                  60                  
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
          65                  70                  75                  80  
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser 
                          85                  90                  95      
          Thr His Val Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100                 105                 110         
          <![CDATA[<210>  129]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A VH CDR1]]>
          <![CDATA[<400>  129]]>
          Ser Gly Tyr Trp Asn 
          1               5   
          <![CDATA[<210>  130]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A VH CDR2]]>
          <![CDATA[<400>  130]]>
          Tyr Ile Ser Asp Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys Ser 
          1               5                   10                  15      
          <![CDATA[<210>  131]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A VH CDR3]]>
          <![CDATA[<400>  131]]>
          Arg Thr Leu Ala Thr Tyr Tyr Ala Met Asp Tyr 
          1               5                   10      
          <![CDATA[<210>  132]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A VL CDR1]]>
          <![CDATA[<400>  132]]>
          Arg Ala Ser Gln Ser Leu Val His Ser Asp Gly Asn Thr Tyr Leu His 
          1               5                   10                  15      
          <![CDATA[<210>  133]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A VL CDR2]]>
          <![CDATA[<400>  133]]>
          Arg Val Ser Asn Arg Phe Ser 
          1               5           
          <![CDATA[<210>  134]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-CD228A VL CDR3]]>
          <![CDATA[<400>  134]]>
          Ser Gln Ser Thr His Val Pro Pro Thr 
          1               5                   
          <![CDATA[<210>  135]]>
          <![CDATA[<211>  120]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV)重鏈可變區]]>
          <![CDATA[<400>  135]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 
          1               5                   10                  15      
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Leu Thr Ile Glu Asp Tyr 
                      20                  25                  30          
          Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met 
                  35                  40                  45              
          Gly Trp Ile Asp Pro Glu Asn Gly Asp Thr Glu Tyr Gly Pro Lys Phe 
              50                  55                  60                  
          Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr 
          65                  70                  75                  80  
          Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Val His Asn Ala His Tyr Gly Thr Trp Phe Ala Tyr Trp Gly Gln 
                      100                 105                 110         
          Gly Thr Leu Val Thr Val Ser Ser 
                  115                 120 
          <![CDATA[<210>  136]]>
          <![CDATA[<211>  112]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV)輕鏈可變區]]>
          <![CDATA[<400>  136]]>
          Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly 
          1               5                   10                  15      
          Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser 
                      20                  25                  30          
          Ser Gly Asn Thr Tyr Leu Glu Trp Tyr Gln Gln Arg Pro Gly Gln Ser 
                  35                  40                  45              
          Pro Arg Pro Leu Ile Tyr Lys Ile Ser Thr Arg Phe Ser Gly Val Pro 
              50                  55                  60                  
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile 
          65                  70                  75                  80  
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly 
                          85                  90                  95      
          Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys 
                      100                 105                 110         
          <![CDATA[<210>  137]]>
          <![CDATA[<211>  5]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV) VH CDR1]]>
          <![CDATA[<400>  137]]>
          Asp Tyr Tyr Met His 
          1               5   
          <![CDATA[<210>  138]]>
          <![CDATA[<211>  17]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV) VH CDR2]]>
          <![CDATA[<400>  138]]>
          Trp Ile Asp Pro Glu Asn Gly Asp Thr Glu Tyr Gly Pro Lys Phe Gln 
          1               5                   10                  15      
          Gly 
          <![CDATA[<210>  139]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV) VH CDR3]]>
          <![CDATA[<400>  139]]>
          His Asn Ala His Tyr Gly Thr Trp Phe Ala Tyr 
          1               5                   10      
          <![CDATA[<210>  140]]>
          <![CDATA[<211>  16]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV) VL CDR1]]>
          <![CDATA[<400>  140]]>
          Arg Ser Ser Gln Ser Leu Leu His Ser Ser Gly Asn Thr Tyr Leu Glu 
          1               5                   10                  15      
          <![CDATA[<210>  141]]>
          <![CDATA[<211>  7]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV) VL CDR2]]>
          <![CDATA[<400>  141]]>
          Lys Ile Ser Thr Arg Phe Ser 
          1               5           
          <![CDATA[<210>  142]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：SGN-LIV1A拉地妥珠單抗維多汀(LV) VL CDR3]]>
          <![CDATA[<400>  142]]>
          Phe Gln Gly Ser His Val Pro Tyr Thr 
          1               5                   
          <![CDATA[<210>  143]]>
          <![CDATA[<211>  118]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV)重鏈可變區]]>
          <![CDATA[<400>  143]]>
          Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly 
          1               5                   10                  15      
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr 
                      20                  25                  30          
          Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val 
                  35                  40                  45              
          Ser Ser Ile Ser Gly Ser Gly Asp Tyr Thr Tyr Tyr Thr Asp Ser Val 
              50                  55                  60                  
          Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr 
          65                  70                  75                  80  
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 
                          85                  90                  95      
          Ala Arg Ser Pro Trp Gly Tyr Tyr Leu Asp Ser Trp Gly Gln Gly Thr 
                      100                 105                 110         
          Leu Val Thr Val Ser Ser 
                  115             
          <![CDATA[<210>  144]]>
          <![CDATA[<211>  107]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV)輕鏈可變區]]>
          <![CDATA[<400>  144]]>
          Asp Ile Gln Met Thr Gln Ser Pro Pro Ser Leu Ser Ala Ser Ala Gly 
          1               5                   10                  15      
          Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Arg 
                      20                  25                  30          
          Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile 
                  35                  40                  45              
          Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly 
              50                  55                  60                  
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro 
          65                  70                  75                  80  
          Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr 
                          85                  90                  95      
          Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys 
                      100                 105         
          <![CDATA[<210>  145]]>
          <![CDATA[<211>  8]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV) VH CDR1]]>
          <![CDATA[<400>  145]]>
          Gly Phe Thr Phe Ser Asn Tyr Ala 
          1               5               
          <![CDATA[<210>  146]]>
          <![CDATA[<211>  8]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV) VH CDR2]]>
          <![CDATA[<400>  146]]>
          Ile Ser Gly Ser Gly Asp Tyr Thr 
          1               5               
          <![CDATA[<210>  147]]>
          <![CDATA[<211>  11]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV) VH CDR3]]>
          <![CDATA[<400>  147]]>
          Ala Arg Ser Pro Trp Gly Tyr Tyr Leu Asp Ser 
          1               5                   10      
          <![CDATA[<210>  148]]>
          <![CDATA[<211>  6]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV) VL CDR1]]>
          <![CDATA[<400>  148]]>
          Gln Gly Ile Ser Ser Arg 
          1               5       
          <![CDATA[<210>  149]]>
          <![CDATA[<211>  3]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV) VL CDR2]]>
          <![CDATA[<400>  149]]>
          Ala Ala Ser 
          1           
          <![CDATA[<210>  150]]>
          <![CDATA[<211>  9]]>
          <![CDATA[<212>  PRT]]>
          <![CDATA[<213>  人工序列]]>
          <![CDATA[<220>]]>
          <![CDATA[<223>  合成：替索圖單抗維多汀(TV) VL CDR3]]>
          <![CDATA[<400>  150]]>
          Gln Gln Tyr Asn Ser Tyr Pro Tyr Thr 
          1               5                   
          
           <![CDATA[ <110> SEAGEN INC.]]>
           <![CDATA[ <120> Combination therapy]]>
           <![CDATA[ <130> 01218-0027-00PCT]]>
           <![CDATA[ <150> US 63/111,045]]>
           <![CDATA[ <151> 2020-11-08]]>
           <![CDATA[ <150> US 63/172,411]]>
           <![CDATA[ <151> 2021-04-08]]>
           <![CDATA[ <150> US 63/208,179]]>
           <![CDATA[ <151> 2021-06-08]]>
           <![CDATA[ <160> 150 ]]>
           <![CDATA[ <170> PatentIn version 3.5]]>
           <![CDATA[ <210> 1]]>
           <![CDATA[ <211> 125]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Anti-TIGIT antibody clone 13 VH protein]]>
           <![CDATA[ <400> 1]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
                  115 120 125
           <![CDATA[ <210> 2]]>
           <![CDATA[ <211> 125]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Anti-TIGIT antibody clone 13A VH]]>
           <![CDATA[ <400> 2]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Leu Ile Pro Tyr Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
                  115 120 125
           <![CDATA[ <210> 3]]>
           <![CDATA[ <211> 125]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Anti-TIGIT antibody clone 13B VH]]>
           <![CDATA[ <400> 3]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ala Trp
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Ser Gly Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
                  115 120 125
           <![CDATA[ <210> 4]]>
           <![CDATA[ <211> 125]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Anti-TIGIT antibody clone 13C VH]]>
           <![CDATA[ <400> 4]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Leu Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
                  115 120 125
           <![CDATA[ <210> 5]]>
           <![CDATA[ <211> 125]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Anti-TIGIT antibody clone 13D VH]]>
           <![CDATA[ <400> 5]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
                  115 120 125
           <![CDATA[ <210> 6]]>
           <![CDATA[ <211> 112]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure lines 13, 13A, 13B, 13C and 13D VL proteins]]>
           <![CDATA[ <400> 6]]>
          Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
          1 5 10 15
          Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
                      20 25 30
          Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser
                  35 40 45
          Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro
              50 55 60
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
          65 70 75 80
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala
                          85 90 95
          Arg Arg Ile Pro Ile Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 7]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13 VH CDR1]]>
           <![CDATA[ <400> 7]]>
          Gly Thr Phe Ser Ser Tyr Ala Ile Ser
          1 5
           <![CDATA[ <210> 8]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure lines 13A, 13C and 13D VH CDR1]]>
           <![CDATA[ <400> 8]]>
          Gly Thr Phe Leu Ser Ser Ala Ile Ser
          1 5
           <![CDATA[ <210> 9]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13B VH CDR1]]>
           <![CDATA[ <400> 9]]>
          Gly Thr Phe Ser Ala Trp Ala Ile Ser
          1 5
           <![CDATA[ <210> 10]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13 VH CDR2]]>
           <![CDATA[ <400> 10]]>
          Ser Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 11]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13A VH CDR2]]>
           <![CDATA[ <400> 11]]>
          Ser Leu Ile Pro Tyr Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 12]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13B and 13D VH CDR2]]>
           <![CDATA[ <400> 12]]>
          Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 13]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13C VH CDR2]]>
           <![CDATA[ <400> 13]]>
          Ser Ile Ile Pro Leu Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 14]]>
           <![CDATA[ <211> 18]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13 and 13A VH CDR3]]>
           <![CDATA[ <400> 14]]>
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe
          1 5 10 15
          Asp Pro
           <![CDATA[ <210> 15]]>
           <![CDATA[ <211> 18]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13B VH CDR3]]>
           <![CDATA[ <400> 15]]>
          Ala Arg Gly Pro Ser Glu Val Ser Gly Ile Leu Gly Tyr Val Trp Phe
          1 5 10 15
          Asp Pro
           <![CDATA[ <210> 16]]>
           <![CDATA[ <211> 18]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13C and 13D VH CDR3]]>
           <![CDATA[ <400> 16]]>
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe
          1 5 10 15
          Asp Pro
           <![CDATA[ <210> 17]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure lines 13, 13A, 13B, 13C and 13D VL CDR1]]>
           <![CDATA[ <400> 17]]>
          Arg Ser Ser Gln Ser Leu Leu His Ser Asn Gly Tyr Asn Tyr Leu Asp
          1 5 10 15
           <![CDATA[ <210> 18]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure lines 13, 13A, 13B, 13C and 13D VL CDR2]]>
           <![CDATA[ <400> 18]]>
          Leu Gly Ser Asn Arg Ala Ser
          1 5
           <![CDATA[ <210> 19]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure lines 13, 13A, 13B, 13C and 13D VL CDR3]]>
           <![CDATA[ <400> 19]]>
          Met Gln Ala Arg Arg Ile Pro Ile Thr
          1 5
           <![CDATA[ <210> 20]]>
           <![CDATA[ <211> 455]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13 heavy chain hIgG1 (and afucosylated hIgG1) amino acid sequence]]>
           <![CDATA[ <400> 20]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ser Tyr
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Ile Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
                  115 120 125
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
              130 135 140
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
          145 150 155 160
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
                          165 170 175
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
                      180 185 190
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
                  195 200 205
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
              210 215 220
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
          225 230 235 240
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
                          245 250 255
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
                      260 265 270
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
                  275 280 285
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
              290 295 300
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
          305 310 315 320
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
                          325 330 335
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
                      340 345 350
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
                  355 360 365
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
              370 375 380
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
          385 390 395 400
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
                          405 410 415
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
                      420 425 430
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
                  435 440 445
          Leu Ser Leu Ser Pro Gly Lys
              450 455
           <![CDATA[ <210> 21]]>
           <![CDATA[ <211> 455]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13A heavy chain hIgG1 (and afucosylated hIgG1) amino acid sequence]]>
           <![CDATA[ <400> 21]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Leu Ile Pro Tyr Phe Gly Thr Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Gly Ala Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
                  115 120 125
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
              130 135 140
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
          145 150 155 160
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
                          165 170 175
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
                      180 185 190
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
                  195 200 205
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
              210 215 220
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
          225 230 235 240
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
                          245 250 255
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
                      260 265 270
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
                  275 280 285
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
              290 295 300
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
          305 310 315 320
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
                          325 330 335
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
                      340 345 350
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
                  355 360 365
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
              370 375 380
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
          385 390 395 400
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
                          405 410 415
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
                      420 425 430
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
                  435 440 445
          Leu Ser Leu Ser Pro Gly Lys
              450 455
           <![CDATA[ <210> 22]]>
           <![CDATA[ <211> 455]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13B heavy chain hIgG1 (and afucosylated hIgG1) amino acid sequence]]>
           <![CDATA[ <400> 22]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Ser Ala Trp
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Ser Gly Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
                  115 120 125
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
              130 135 140
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
          145 150 155 160
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
                          165 170 175
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
                      180 185 190
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
                  195 200 205
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
              210 215 220
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
          225 230 235 240
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
                          245 250 255
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
                      260 265 270
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
                  275 280 285
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
              290 295 300
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
          305 310 315 320
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
                          325 330 335
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
                      340 345 350
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
                  355 360 365
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
              370 375 380
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
          385 390 395 400
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
                          405 410 415
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
                      420 425 430
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
                  435 440 445
          Leu Ser Leu Ser Pro Gly Lys
              450 455
           <![CDATA[ <210> 23]]>
           <![CDATA[ <211> 455]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure line 13C heavy chain hIgG1 (and afucosylated hIgG1) amino acid sequence]]>
           <![CDATA[ <400> 23]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Leu Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
                  115 120 125
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
              130 135 140
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
          145 150 155 160
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
                          165 170 175
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
                      180 185 190
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
                  195 200 205
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
              210 215 220
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
          225 230 235 240
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
                          245 250 255
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
                      260 265 270
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
                  275 280 285
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
              290 295 300
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
          305 310 315 320
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
                          325 330 335
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
                      340 345 350
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
                  355 360 365
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
              370 375 380
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
          385 390 395 400
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
                          405 410 415
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
                      420 425 430
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
                  435 440 445
          Leu Ser Leu Ser Pro Gly Lys
              450 455
           <![CDATA[ <210> 24]]>
           <![CDATA[ <211> 455]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure 13D heavy chain hIgG1 (and afucosylated hIgG1) amino acid sequence]]>
           <![CDATA[ <400> 24]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Gly Thr Phe Leu Ser Ser
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Ser Ile Ile Pro Tyr Phe Gly Lys Ala Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Gly Pro Ser Glu Val Lys Gly Ile Leu Gly Tyr Val Trp Phe
                      100 105 110
          Asp Pro Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr
                  115 120 125
          Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser
              130 135 140
          Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu
          145 150 155 160
          Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His
                          165 170 175
          Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser
                      180 185 190
          Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys
                  195 200 205
          Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu
              210 215 220
          Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
          225 230 235 240
          Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
                          245 250 255
          Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
                      260 265 270
          Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
                  275 280 285
          Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
              290 295 300
          Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
          305 310 315 320
          Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu
                          325 330 335
          Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
                      340 345 350
          Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
                  355 360 365
          Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
              370 375 380
          Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
          385 390 395 400
          Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
                          405 410 415
          Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
                      420 425 430
          Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
                  435 440 445
          Leu Ser Leu Ser Pro Gly Lys
              450 455
           <![CDATA[ <210> 25]]>
           <![CDATA[ <211> 219]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: pure lines 13, 13A, 13B, 13C and 13D light chain hκ (and afucosylated)]]>
                 amino acid sequence
           <![CDATA[ <400> 25]]>
          Asp Ile Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Pro Gly
          1 5 10 15
          Glu Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
                      20 25 30
          Asn Gly Tyr Asn Tyr Leu Asp Trp Tyr Leu Gln Lys Pro Gly Gln Ser
                  35 40 45
          Pro Gln Leu Leu Ile Tyr Leu Gly Ser Asn Arg Ala Ser Gly Val Pro
              50 55 60
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
          65 70 75 80
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Met Gln Ala
                          85 90 95
          Arg Arg Ile Pro Ile Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100 105 110
          Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
                  115 120 125
          Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
              130 135 140
          Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
          145 150 155 160
          Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
                          165 170 175
          Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
                      180 185 190
          Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
                  195 200 205
          Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
              210 215
           <![CDATA[ <210> 26]]>
           <![CDATA[ <211> 444]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 (non-fucosylated hS2C6) heavy chain]]>
           <![CDATA[ <400> 26]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
          1 5 10 15
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
                      20 25 30
          Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
                  35 40 45
          Ala Arg Val Ile Pro Asn Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe
              50 55 60
          Lys Gly Arg Phe Thr Leu Ser Val Asp Asn Ser Lys Asn Thr Ala Tyr
          65 70 75 80
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Glu Gly Ile Tyr Trp Trp Gly Gln Gly Thr Leu Val Thr Val
                      100 105 110
          Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser
                  115 120 125
          Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys
              130 135 140
          Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu
          145 150 155 160
          Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly Leu
                          165 170 175
          Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr
                      180 185 190
          Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val
                  195 200 205
          Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro
              210 215 220
          Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe
          225 230 235 240
          Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val
                          245 250 255
          Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe
                      260 265 270
          Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro
                  275 280 285
          Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr
              290 295 300
          Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val
          305 310 315 320
          Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala
                          325 330 335
          Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg
                      340 345 350
          Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly
                  355 360 365
          Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro
              370 375 380
          Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser
          385 390 395 400
          Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln
                          405 410 415
          Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His
                      420 425 430
          Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
                  435 440
           <![CDATA[ <210> 27]]>
           <![CDATA[ <211> 219]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 (Afucosylated hS2C6) Light Chain]]>
           <![CDATA[ <400> 27]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Ser Leu Val His Ser
                      20 25 30
          Asn Gly Asn Thr Phe Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala
                  35 40 45
          Pro Lys Leu Leu Ile Tyr Thr Val Ser Asn Arg Phe Ser Gly Val Pro
              50 55 60
          Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
          65 70 75 80
          Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Ser Gln Thr
                          85 90 95
          Thr His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
                      100 105 110
          Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
                  115 120 125
          Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
              130 135 140
          Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
          145 150 155 160
          Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
                          165 170 175
          Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
                      180 185 190
          Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
                  195 200 205
          Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
              210 215
           <![CDATA[ <210> 28]]>
           <![CDATA[ <211> 114]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: VH of SEA-CD40]]>
           <![CDATA[ <400> 28]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
          1 5 10 15
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Ser Phe Thr Gly Tyr
                      20 25 30
          Tyr Ile His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
                  35 40 45
          Ala Arg Val Ile Pro Asn Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe
              50 55 60
          Lys Gly Arg Phe Thr Leu Ser Val Asp Asn Ser Lys Asn Thr Ala Tyr
          65 70 75 80
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Glu Gly Ile Tyr Trp Trp Gly Gln Gly Thr Leu Val Thr Val
                      100 105 110
          Ser Ser
           <![CDATA[ <210> 29]]>
           <![CDATA[ <211> 112]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: VL of SEA-CD40]]>
           <![CDATA[ <400> 29]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Gln Ser Leu Val His Ser
                      20 25 30
          Asn Gly Asn Thr Phe Leu His Trp Tyr Gln Gln Lys Pro Gly Lys Ala
                  35 40 45
          Pro Lys Leu Leu Ile Tyr Thr Val Ser Asn Arg Phe Ser Gly Val Pro
              50 55 60
          Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
          65 70 75 80
          Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Phe Cys Ser Gln Thr
                          85 90 95
          Thr His Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 30]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 VH CDR1]]>
           <![CDATA[ <400> 30]]>
          Gly Tyr Tyr Ile His
          1 5
           <![CDATA[ <210> 31]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 VH CDR2]]>
           <![CDATA[ <400> 31]]>
          Arg Val Ile Pro Asn Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 32]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 VH CDR3]]>
           <![CDATA[ <400> 32]]>
          Glu Gly Ile Tyr Trp
          1 5
           <![CDATA[ <210> 33]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 VL CDR1]]>
           <![CDATA[ <400> 33]]>
          Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Phe Leu His
          1 5 10 15
           <![CDATA[ <210> 34]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 VL CDR2]]>
           <![CDATA[ <400> 34]]>
          Thr Val Ser Asn Arg Phe Ser
          1 5
           <![CDATA[ <210> 35]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD40 VL CDR3]]>
           <![CDATA[ <400> 35]]>
          Ser Gln Thr Thr His Val Pro Trp Thr
          1 5
           <![CDATA[ <210> 36]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Alternative anti-CD40 antibody CDR2]]>
           <![CDATA[ <400> 36]]>
          Arg Val Ile Pro Gln Ala Gly Gly Thr Ser Tyr Asn Gln Lys Phe Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 37]]>
           <![CDATA[ <211> 116]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B6A heavy chain variable region]]>
           <![CDATA[ <400> 37]]>
          Gln Phe Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr
                      20 25 30
          Asn Val Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
                  35 40 45
          Gly Val Ile Asn Pro Lys Tyr Gly Thr Thr Arg Tyr Asn Gln Lys Phe
              50 55 60
          Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Thr Arg Gly Leu Asn Ala Trp Asp Tyr Trp Gly Gln Gly Thr Leu Val
                      100 105 110
          Thr Val Ser Ser
                  115
           <![CDATA[ <210> 38]]>
           <![CDATA[ <211> 107]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B6A light chain variable region]]>
           <![CDATA[ <400> 38]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Ala
                      20 25 30
          Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
                  35 40 45
          Tyr Gly Ala Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Arg Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Asn Val Leu Thr Thr Pro Tyr
                          85 90 95
          Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100 105
           <![CDATA[ <210> 39]]>
           <![CDATA[ <211> 115]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-STNV heavy chain variable region]]>
           <![CDATA[ <400> 39]]>
          Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp His
                      20 25 30
          Ala Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Tyr Phe Ser Pro Gly Asn Asp Asp Ile Lys Tyr Asn Glu Lys Phe
              50 55 60
          Arg Gly Arg Val Thr Met Thr Ala Asp Lys Ser Ser Ser Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Arg Ser Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
                          85 90 95
          Lys Arg Ser Leu Ser Thr Pro Tyr Trp Gly Gln Gly Thr Leu Val Thr
                      100 105 110
          Val Ser Ser
                  115
           <![CDATA[ <210> 40]]>
           <![CDATA[ <211> 113]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-STNV heavy chain variable region]]>
           <![CDATA[ <400> 40]]>
          Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
          1 5 10 15
          Glu Arg Ala Thr Ile Asn Cys Lys Ser Ser Gln Ser Leu Leu Asn Arg
                      20 25 30
          Gly Asn His Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
                  35 40 45
          Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
              50 55 60
          Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
          65 70 75 80
          Ile Ser Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln Asn
                          85 90 95
          Asp Tyr Thr Tyr Pro Tyr Thr Phe Gly Gln Gly Thr Lys Val Glu Ile
                      100 105 110
          Lys
           <![CDATA[ <210> 41]]>
           <![CDATA[ <211> 118]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 heavy chain variable region]]>
           <![CDATA[ <400> 41]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
                      20 25 30
          Gly Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Lys Trp Met
                  35 40 45
          Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe
              50 55 60
          Lys Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Asp Tyr Gly Asp Tyr Gly Met Asp Tyr Trp Gly Gln Gly Thr
                      100 105 110
          Thr Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 42]]>
           <![CDATA[ <211> 111]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD70 light chain variable region]]>
           <![CDATA[ <400> 42]]>
          Asp Ile Val Met Thr Gln Ser Pro Asp Ser Leu Ala Val Ser Leu Gly
          1 5 10 15
          Glu Arg Ala Thr Ile Asn Cys Arg Ala Ser Lys Ser Val Ser Thr Ser
                      20 25 30
          Gly Tyr Ser Phe Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro
                  35 40 45
          Lys Leu Leu Ile Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Asp
              50 55 60
          Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser
          65 70 75 80
          Ser Leu Gln Ala Glu Asp Val Ala Val Tyr Tyr Cys Gln His Ser Arg
                          85 90 95
          Glu Val Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 43]]>
           <![CDATA[ <211> 119]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A heavy chain variable region]]>
           <![CDATA[ <400> 43]]>
          Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
          1 5 10 15
          Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Thr Ser Gly
                      20 25 30
          Tyr Trp Asn Trp Ile Arg Gln Pro Gly Lys Gly Leu Glu Tyr Ile
                  35 40 45
          Gly Tyr Ile Ser Asp Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys
              50 55 60
          Ser Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Ser Leu
          65 70 75 80
          Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala
                          85 90 95
          Arg Arg Thr Leu Ala Thr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly
                      100 105 110
          Thr Leu Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 44]]>
           <![CDATA[ <211> 112]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A light chain variable region]]>
           <![CDATA[ <400> 44]]>
          Asp Phe Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
          1 5 10 15
          Gln Pro Ala Ser Ile Ser Cys Arg Ala Ser Gln Ser Leu Val His Ser
                      20 25 30
          Asp Gly Asn Thr Tyr Leu His Trp Tyr Gln Gln Arg Pro Gly Gln Ser
                  35 40 45
          Pro Arg Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro
              50 55 60
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
          65 70 75 80
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser
                          85 90 95
          Thr His Val Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 45]]>
           <![CDATA[ <211> 121]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA heavy chain variable region]]>
           <![CDATA[ <400> 45]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr
                      20 25 30
          Tyr Ile His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
                  35 40 45
          Gly Tyr Ile Asn Pro Asn Ser Gly Tyr Thr Asn Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Ala Thr Met Thr Ala Asp Lys Ser Ile Asn Thr Ala Tyr
          65 70 75 80
          Val Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Phe Cys
                          85 90 95
          Thr Arg Tyr Met Trp Glu Arg Val Thr Gly Phe Phe Asp Phe Trp Gly
                      100 105 110
          Gln Gly Thr Met Val Thr Val Ser Ser
                  115 120
           <![CDATA[ <210> 46]]>
           <![CDATA[ <211> 107]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA light chain variable region]]>
           <![CDATA[ <400> 46]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Leu Ala Ser Glu Asp Ile Ser Asp Asp
                      20 25 30
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Val Leu Val
                  35 40 45
          Tyr Thr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Phe Ala Thr Tyr Phe Cys Gln Gln Thr Tyr Lys Phe Pro Pro
                          85 90 95
          Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100 105
           <![CDATA[ <210> 47]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA VH CDR1]]>
           <![CDATA[ <400> 47]]>
          Asp Tyr Tyr Ile His
          1 5
           <![CDATA[ <210> 48]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA VH CDR2]]>
           <![CDATA[ <400> 48]]>
          Tyr Ile Asn Pro Asn Ser Gly Tyr Thr Asn Tyr Ala Gln Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 49]]>
           <![CDATA[ <211> 12]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA VH CDR3]]>
           <![CDATA[ <400> 49]]>
          Tyr Met Trp Glu Arg Val Thr Gly Phe Phe Asp Phe
          1 5 10
           <![CDATA[ <210> 50]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA VL CDR1]]>
           <![CDATA[ <400> 50]]>
          Leu Ala Ser Glu Asp Ile Ser Asp Asp Leu Ala
          1 5 10
           <![CDATA[ <210> 51]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA VL CDR2]]>
           <![CDATA[ <400> 51]]>
          Thr Thr Ser Ser Leu Gln Ser
          1 5
           <![CDATA[ <210> 52]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-BCMA VL CDR3]]>
           <![CDATA[ <400> 52]]>
          Gln Gln Thr Tyr Lys Phe Pro Pro Thr
          1 5
           <![CDATA[ <210> 53]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 VH CDR1]]>
           <![CDATA[ <400> 53]]>
          Asn Tyr Gly Met Asn
          1 5
           <![CDATA[ <210> 54]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 VH CDR2]]>
           <![CDATA[ <400> 54]]>
          Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Asp Ala Phe Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 55]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 VH CDR3]]>
           <![CDATA[ <400> 55]]>
          Asp Tyr Gly Asp Tyr Gly Met Asp Tyr
          1 5
           <![CDATA[ <210> 56]]>
           <![CDATA[ <211> 15]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 VL CDR1]]>
           <![CDATA[ <400> 56]]>
          Arg Ala Ser Lys Ser Val Ser Thr Ser Gly Tyr Ser Phe Met His
          1 5 10 15
           <![CDATA[ <210> 57]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 VL CDR2]]>
           <![CDATA[ <400> 57]]>
          Leu Ala Ser Asn Leu Glu Ser
          1 5
           <![CDATA[ <210> 58]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SEA-CD70 VL CDR3]]>
           <![CDATA[ <400> 58]]>
          Gln His Ser Arg Glu Val Pro Trp Thr
          1 5
           <![CDATA[ <210> 59]]>
           <![CDATA[ <211> 118]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuximab (175D10) heavy chain variable region]]>
           <![CDATA[ <400> 59]]>
          Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala
          1 5 10 15
          Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Ser Tyr
                      20 25 30
          Trp Ile Asn Trp Val Lys Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile
                  35 40 45
          Gly Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe
              50 55 60
          Lys Asp Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Ser Thr Ala Tyr
          65 70 75 80
          Met Gln Leu Ser Ser Pro Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys
                          85 90 95
          Thr Arg Ser Trp Arg Gly Asn Ser Phe Asp Tyr Trp Gly Gln Gly Thr
                      100 105 110
          Thr Leu Thr Val Ser Ser
                  115
           <![CDATA[ <210> 60]]>
           <![CDATA[ <211> 109]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuximab (175D10) light chain variable region]]>
           <![CDATA[ <400> 60]]>
          Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly
          1 5 10 15
          Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
                      20 25 30
          Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
                  35 40 45
          Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
              50 55 60
          Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
          65 70 75 80
          Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
                          85 90 95
          Asp Tyr Ser Tyr Pro Phe Thr Phe Gly Ser Gly Thr Lys
                      100 105
           <![CDATA[ <210> 61]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuzumab (175D10) VH CDR1]]>
           <![CDATA[ <400> 61]]>
          Ser Tyr Trp Ile Asn
          1 5
           <![CDATA[ <210> 62]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuzumab (175D10) VH CDR2]]>
           <![CDATA[ <400> 62]]>
          Asn Ile Tyr Pro Ser Asp Ser Tyr Thr Asn Tyr Asn Gln Lys Phe Lys
          1 5 10 15
          Asp
           <![CDATA[ <210> 63]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuzumab (175D10) VH CDR3]]>
           <![CDATA[ <400> 63]]>
          Ser Trp Arg Gly Asn Ser Phe Asp Tyr
          1 5
           <![CDATA[ <210> 64]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuzumab (175D10) VL CDR1]]>
           <![CDATA[ <400> 64]]>
          Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu
          1 5 10 15
          Thr
           <![CDATA[ <210> 65]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuzumab (175D10) VL CDR2]]>
           <![CDATA[ <400> 65]]>
          Trp Ala Ser Thr Arg Glu Ser
          1 5
           <![CDATA[ <210> 66]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Zobetuzumab (175D10) VL CDR3]]>
           <![CDATA[ <400> 66]]>
          Gln Asn Asp Tyr Ser Tyr Pro Phe Thr
          1 5
           <![CDATA[ <210> 67]]>
           <![CDATA[ <211> 118]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 heavy chain variable region]]>
           <![CDATA[ <400> 67]]>
          Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu
          1 5 10 15
          Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asn Tyr
                      20 25 30
          Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu Lys Trp Met
                  35 40 45
          Gly Trp Ile Asn Thr Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe
              50 55 60
          Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Ser Thr Ala Tyr
          65 70 75 80
          Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp Thr Ala Thr Tyr Phe Cys
                          85 90 95
          Ala Arg Leu Gly Phe Gly Asn Ala Met Asp Tyr Trp Gly Gln Gly Thr
                      100 105 110
          Ser Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 68]]>
           <![CDATA[ <211> 113]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 light chain variable region]]>
           <![CDATA[ <400> 68]]>
          Asp Ile Val Met Thr Gln Ser Pro Ser Ser Leu Thr Val Thr Ala Gly
          1 5 10 15
          Glu Lys Val Thr Met Ser Cys Lys Ser Ser Gln Ser Leu Leu Asn Ser
                      20 25 30
          Gly Asn Gln Lys Asn Tyr Leu Thr Trp Tyr Gln Gln Lys Pro Gly Gln
                  35 40 45
          Pro Pro Lys Leu Leu Ile Tyr Trp Ala Ser Thr Arg Glu Ser Gly Val
              50 55 60
          Pro Asp Arg Phe Thr Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr
          65 70 75 80
          Ile Ser Ser Val Gln Ala Glu Asp Leu Ala Val Tyr Tyr Cys Gln Asn
                          85 90 95
          Asp Tyr Ser Tyr Pro Leu Thr Phe Gly Ala Gly Thr Lys Leu Glu Leu
                      100 105 110
          Lys
           <![CDATA[ <210> 69]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 VH CDR1]]>
           <![CDATA[ <400> 69]]>
          Asn Tyr Gly Met Asn
          1 5
           <![CDATA[ <210> 70]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 VH CDR2]]>
           <![CDATA[ <400> 70]]>
          Trp Ile Asn Thr Asn Thr Gly Glu Pro Thr Tyr Ala Glu Glu Phe Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 71]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 VH CDR3]]>
           <![CDATA[ <400> 71]]>
          Leu Gly Phe Gly Asn Ala Met Asp Tyr
          1 5
           <![CDATA[ <210> 72]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 VL CDR1]]>
           <![CDATA[ <400> 72]]>
          Lys Ser Ser Gln Ser Leu Leu Asn Ser Gly Asn Gln Lys Asn Tyr Leu
          1 5 10 15
          Thr
           <![CDATA[ <210> 73]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 VL CDR2]]>
           <![CDATA[ <400> 73]]>
          Trp Ala Ser Thr Arg Glu Ser
          1 5
           <![CDATA[ <210> 74]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: 163E12 VL CDR3]]>
           <![CDATA[ <400> 74]]>
          Gln Asn Asp Tyr Ser Tyr Pro Leu Thr
          1 5
           <![CDATA[ <210> 75]]>
           <![CDATA[ <211> 123]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V heavy chain variable region]]>
           <![CDATA[ <400> 75]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ser
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Thr Ser Gly Asp Thr Phe Ser Thr Ala
                      20 25 30
          Ala Ile Ser Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Gly Ile Ile Pro Ile Phe Gly Lys Ala His Tyr Ala Gln Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Ile Thr Ala Asp Glu Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
                          85 90 95
          Ala Arg Lys Phe His Phe Val Ser Gly Ser Pro Phe Gly Met Asp Val
                      100 105 110
          Trp Gly Gln Gly Thr Thr Val Thr Val Ser Ser
                  115 120
           <![CDATA[ <210> 76]]>
           <![CDATA[ <211> 106]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V light chain variable region]]>
           <![CDATA[ <400> 76]]>
          Glu Ile Val Leu Thr Gln Ser Pro Ala Thr Leu Ser Leu Ser Pro Gly
          1 5 10 15
          Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Tyr
                      20 25 30
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
                  35 40 45
          Tyr Asp Ala Ser Asn Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Glu Pro
          65 70 75 80
          Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Arg Ser Asn Trp Pro Thr
                          85 90 95
          Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
                      100 105
           <![CDATA[ <210> 77]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V VH CDR1]]>
           <![CDATA[ <400> 77]]>
          Thr Ala Ala Ile Ser
          1 5
           <![CDATA[ <210> 78]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V VH CDR2]]>
           <![CDATA[ <400> 78]]>
          Gly Ile Ile Pro Ile Phe Gly Lys Ala His Tyr Ala Gln Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 79]]>
           <![CDATA[ <211> 14]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V VH CDR3]]>
           <![CDATA[ <400> 79]]>
          Lys Phe His Phe Val Ser Gly Ser Pro Phe Gly Met Asp Val
          1 5 10
           <![CDATA[ <210> 80]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V VL CDR1]]>
           <![CDATA[ <400> 80]]>
          Arg Ala Ser Gln Ser Val Ser Ser Tyr Leu Ala
          1 5 10
           <![CDATA[ <210> 81]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V VL CDR2]]>
           <![CDATA[ <400> 81]]>
          Asp Ala Ser Asn Arg Ala Thr
          1 5
           <![CDATA[ <210> 82]]>
           <![CDATA[ <211> 8]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-PDL1V VL CDR3]]>
           <![CDATA[ <400> 82]]>
          Gln Gln Arg Ser Asn Trp Pro Thr
          1 5
           <![CDATA[ <210> 83]]>
           <![CDATA[ <211> 123]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV heavy chain variable region]]>
           <![CDATA[ <400> 83]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Arg
          1 5 10 15
          Ser Leu Arg Leu Ser Cys Thr Ala Ser Gly Phe Thr Phe Thr Asp Tyr
                      20 25 30
          Tyr Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Leu
                  35 40 45
          Ala Leu Ile Arg Asn Lys Ala Thr Gly Tyr Thr Thr Glu Tyr Thr Ala
              50 55 60
          Ser Val Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Ser Ile
          65 70 75 80
          Leu Tyr Leu Gln Met Asn Ser Leu Lys Thr Glu Asp Thr Ala Val Tyr
                          85 90 95
          Tyr Cys Ala Arg Ala Ser Phe Tyr Tyr Asp Gly Lys Val Leu Ala Tyr
                      100 105 110
          Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
                  115 120
           <![CDATA[ <210> 84]]>
           <![CDATA[ <211> 106]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV light chain variable region]]>
           <![CDATA[ <400> 84]]>
          Asp Thr Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Gln Ala Ser Gln Asp Ile Asn Lys Tyr
                      20 25 30
          Leu Ala Trp Tyr Gln Tyr Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
                  35 40 45
          His Tyr Thr Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Arg Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Ile Ala Thr Tyr Tyr Cys Leu Gln Tyr Asp Asn Leu Tyr Thr
                          85 90 95
          Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100 105
           <![CDATA[ <210> 85]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV VH CDR1]]>
           <![CDATA[ <400> 85]]>
          Asp Tyr Tyr Met Ser
          1 5
           <![CDATA[ <210> 86]]>
           <![CDATA[ <211> 19]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV VH CDR2]]>
           <![CDATA[ <400> 86]]>
          Leu Ile Arg Asn Lys Ala Thr Gly Tyr Thr Thr Glu Tyr Thr Ala Ser
          1 5 10 15
          Val Lys Gly
           <![CDATA[ <210> 87]]>
           <![CDATA[ <211> 12]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV VH CDR3]]>
           <![CDATA[ <400> 87]]>
          Ala Ser Phe Tyr Tyr Asp Gly Lys Val Leu Ala Tyr
          1 5 10
           <![CDATA[ <210> 88]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV VL CDR1]]>
           <![CDATA[ <400> 88]]>
          Gln Ala Ser Gln Asp Ile Asn Lys Tyr Leu Ala
          1 5 10
           <![CDATA[ <210> 89]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV VL CDR2]]>
           <![CDATA[ <400> 89]]>
          Tyr Thr Ser Ser Leu Gln Ser
          1 5
           <![CDATA[ <210> 90]]>
           <![CDATA[ <211> 8]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-ALPV VL CDR3]]>
           <![CDATA[ <400> 90]]>
          Leu Gln Tyr Asp Asn Leu Tyr Thr
          1 5
           <![CDATA[ <210> 91]]>
           <![CDATA[ <211> 120]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V heavy chain variable region]]>
           <![CDATA[ <400> 91]]>
          Gln Leu Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
          1 5 10 15
          Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Gly Ser Ile Lys Ser Gly
                      20 25 30
          Ser Tyr Tyr Trp Gly Trp Ile Arg Gln Pro Pro Gly Lys Gly Leu Glu
                  35 40 45
          Trp Ile Gly Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser
              50 55 60
          Leu Arg Ser Arg Val Thr Ile Ser Val Asp Thr Ser Lys Asn Gln Phe
          65 70 75 80
          Ser Leu Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr
                          85 90 95
          Cys Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro Trp Gly Gln
                      100 105 110
          Gly Thr Leu Val Thr Val Ser Ser
                  115 120
           <![CDATA[ <210> 92]]>
           <![CDATA[ <211> 107]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V light chain variable region]]>
           <![CDATA[ <400> 92]]>
          Glu Ile Val Met Thr Gln Ser Pro Ala Thr Leu Ser Val Ser Pro Gly
          1 5 10 15
          Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Asn
                      20 25 30
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile
                  35 40 45
          Tyr Gly Ala Ser Thr Arg Ala Thr Gly Ile Pro Ala Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Thr Glu Phe Thr Leu Thr Ile Ser Ser Leu Gln Ser
          65 70 75 80
          Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr His Ser Phe Pro Phe
                          85 90 95
          Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100 105
           <![CDATA[ <210> 93]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V VH CDR1]]>
           <![CDATA[ <400> 93]]>
          Gly Ser Ile Lys Ser Gly Ser Tyr Tyr Trp Gly
          1 5 10
           <![CDATA[ <210> 94]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V VH CDR2]]>
           <![CDATA[ <400> 94]]>
          Asn Ile Tyr Tyr Ser Gly Ser Thr Tyr Tyr Asn Pro Ser Leu Arg Ser
          1 5 10 15
           <![CDATA[ <210> 95]]>
           <![CDATA[ <211> 12]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V VH CDR3]]>
           <![CDATA[ <400> 95]]>
          Ala Arg Glu Gly Ser Tyr Pro Asn Gln Phe Asp Pro
          1 5 10
           <![CDATA[ <210> 96]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V VL CDR1]]>
           <![CDATA[ <400> 96]]>
          Arg Ala Ser Gln Ser Val Ser Ser Asn Leu Ala
          1 5 10
           <![CDATA[ <210> 97]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V VL CDR2]]>
           <![CDATA[ <400> 97]]>
          Gly Ala Ser Thr Arg Ala Thr
          1 5
           <![CDATA[ <210> 98]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-B7H4V VL CDR3]]>
           <![CDATA[ <400> 98]]>
          Gln Gln Tyr His Ser Phe Pro Phe Thr
          1 5
           <![CDATA[ <210> 99]]>
           <![CDATA[ <211> 445]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: dicituzumab vedotin heavy chain]]>
           <![CDATA[ <400> 99]]>
          Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Thr Val Lys Ile Ser Cys Lys Val Ser Gly Tyr Thr Phe Thr Asp Tyr
                      20 25 30
          Tyr Ile His Trp Val Gln Gln Ala Pro Gly Lys Gly Leu Glu Trp Met
                  35 40 45
          Gly Arg Val Asn Pro Asp His Gly Asp Ser Tyr Tyr Asn Gln Lys Phe
              50 55 60
          Lys Asp Lys Ala Thr Ile Thr Ala Asp Lys Ser Thr Asp Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Phe Cys
                          85 90 95
          Ala Arg Asn Tyr Leu Phe Asp His Trp Gly Gln Gly Thr Leu Val Thr
                      100 105 110
          Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro
                  115 120 125
          Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val
              130 135 140
          Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala
          145 150 155 160
          Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu Gln Ser Ser Gly
                          165 170 175
          Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly
                      180 185 190
          Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys
                  195 200 205
          Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys
              210 215 220
          Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu
          225 230 235 240
          Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro Glu
                          245 250 255
          Val Thr Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys
                      260 265 270
          Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys
                  275 280 285
          Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu
              290 295 300
          Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys
          305 310 315 320
          Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu Lys Thr Ile Ser Lys
                          325 330 335
          Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser
                      340 345 350
          Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val Lys
                  355 360 365
          Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly Gln
              370 375 380
          Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly
          385 390 395 400
          Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gln
                          405 410 415
          Gln Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn
                      420 425 430
          His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro Gly Lys
                  435 440 445
           <![CDATA[ <210> 100]]>
           <![CDATA[ <211> 212]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: dicituzumab vedotin light chain]]>
           <![CDATA[ <400> 100]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Lys Ala Ser Gln Asp Val Gly Thr Ala
                      20 25 30
          Val Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
                  35 40 45
          Tyr Trp Ala Ser Ile Arg His Thr Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Phe Ala Thr Tyr Tyr Cys His Gln Phe Ala Thr Tyr Thr Phe
                          85 90 95
          Gly Gly Gly Thr Lys Val Glu Ile Lys Arg Thr Val Ala Ala Pro Ser
                      100 105 110
          Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly Thr Ala
                  115 120 125
          Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val
              130 135 140
          Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln Glu Ser
          145 150 155 160
          Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr
                          165 170 175
          Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys
                      180 185 190
          Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn
                  195 200 205
          Arg Gly Glu Cys
              210
           <![CDATA[ <210> 101]]>
           <![CDATA[ <211> 450]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Rivatuzumab vedotin heavy chain]]>
           <![CDATA[ <400> 101]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
          1 5 10 15
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Ser Phe Ser Asp Phe
                      20 25 30
          Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
                  35 40 45
          Ala Thr Ile Gly Arg Val Ala Phe His Thr Tyr Tyr Pro Asp Ser Met
              50 55 60
          Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
          65 70 75 80
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg His Arg Gly Phe Asp Val Gly His Phe Asp Phe Trp Gly Gln
                      100 105 110
          Gly Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val
                  115 120 125
          Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser Gly Gly Thr Ala Ala
              130 135 140
          Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser
          145 150 155 160
          Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val
                          165 170 175
          Leu Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro
                      180 185 190
          Ser Ser Ser Leu Gly Thr Gln Thr Tyr Ile Cys Asn Val Asn His Lys
                  195 200 205
          Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu Pro Lys Ser Cys Asp
              210 215 220
          Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro Glu Leu Leu Gly Gly
          225 230 235 240
          Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile
                          245 250 255
          Ser Arg Thr Pro Glu Val Thr Cys Val Val Val Asp Val Ser His Glu
                      260 265 270
          Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp Gly Val Glu Val His
                  275 280 285
          Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr Asn Ser Thr Tyr Arg
              290 295 300
          Val Val Ser Val Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys
          305 310 315 320
          Glu Tyr Lys Cys Lys Val Ser Asn Lys Ala Leu Pro Ala Pro Ile Glu
                          325 330 335
          Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr
                      340 345 350
          Thr Leu Pro Pro Ser Arg Glu Glu Met Thr Lys Asn Gln Val Ser Leu
                  355 360 365
          Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp
              370 375 380
          Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val
          385 390 395 400
          Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser Lys Leu Thr Val Asp
                          405 410 415
          Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser Cys Ser Val Met His
                      420 425 430
          Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Pro
                  435 440 445
          Gly Lys
              450
           <![CDATA[ <210> 102]]>
           <![CDATA[ <211> 219]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: Rivatuzumab vedotin light chain]]>
           <![CDATA[ <400> 102]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Arg Ser Ser Glu Thr Leu Val His Ser
                      20 25 30
          Ser Gly Asn Thr Tyr Leu Glu Trp Tyr Gln Gln Lys Pro Gly Lys Ala
                  35 40 45
          Pro Lys Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro
              50 55 60
          Ser Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile
          65 70 75 80
          Ser Ser Leu Gln Pro Glu Asp Phe Ala Thr Tyr Tyr Cys Phe Gln Gly
                          85 90 95
          Ser Phe Asn Pro Leu Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
                      100 105 110
          Arg Thr Val Ala Ala Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu
                  115 120 125
          Gln Leu Lys Ser Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe
              130 135 140
          Tyr Pro Arg Glu Ala Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln
          145 150 155 160
          Ser Gly Asn Ser Gln Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser
                          165 170 175
          Thr Tyr Ser Leu Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu
                      180 185 190
          Lys His Lys Val Tyr Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser
                  195 200 205
          Pro Val Thr Lys Ser Phe Asn Arg Gly Glu Cys
              210 215
           <![CDATA[ <210> 103]]>
           <![CDATA[ <211> 117]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) heavy chain variable region]]>
           <![CDATA[ <400> 103]]>
          Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
          1 5 10 15
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Ser Tyr
                      20 25 30
          Asn Met Asn Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
                  35 40 45
          Ser Tyr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val
              50 55 60
          Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Ser
          65 70 75 80
          Leu Gln Met Asn Ser Leu Arg Asp Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Ala Tyr Tyr Tyr Gly Met Asp Val Trp Gly Gln Gly Thr Thr
                      100 105 110
          Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 104]]>
           <![CDATA[ <211> 107]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) light chain variable region]]>
           <![CDATA[ <400> 104]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Gly Trp
                      20 25 30
          Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Phe Leu Ile
                  35 40 45
          Tyr Ala Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ser Phe Pro Pro
                          85 90 95
          Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100 105
           <![CDATA[ <210> 105]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) VH CDR1]]>
           <![CDATA[ <400> 105]]>
          Ser Tyr Asn Met Asn
          1 5
           <![CDATA[ <210> 106]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) VH CDR2]]>
           <![CDATA[ <400> 106]]>
          Tyr Ile Ser Ser Ser Ser Ser Thr Ile Tyr Tyr Ala Asp Ser Val Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 107]]>
           <![CDATA[ <211> 8]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) VH CDR3]]>
           <![CDATA[ <400> 107]]>
          Ala Tyr Tyr Tyr Gly Met Asp Val
          1 5
           <![CDATA[ <210> 108]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) VL CDR1]]>
           <![CDATA[ <400> 108]]>
          Arg Ala Ser Gln Gly Ile Ser Gly Trp Leu Ala
          1 5 10
           <![CDATA[ <210> 109]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) VL CDR2]]>
           <![CDATA[ <400> 109]]>
          Ala Ala Ser Thr Leu Gln Ser
          1 5
           <![CDATA[ <210> 110]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: PADCEV (ennozumab vedotin) (EV) VL CDR3]]>
           <![CDATA[ <400> 110]]>
          Gln Gln Ala Asn Ser Phe Pro Pro Thr
          1 5
           <![CDATA[ <210> 111]]>
           <![CDATA[ <211> 116]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 heavy chain variable region]]>
           <![CDATA[ <400> 111]]>
          Gln Phe Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Asp Tyr
                      20 25 30
          Asn Val Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile
                  35 40 45
          Gly Val Ile Asn Pro Lys Tyr Gly Thr Thr Arg Tyr Asn Gln Lys Phe
              50 55 60
          Lys Gly Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Thr Arg Gly Leu Asn Ala Trp Asp Tyr Trp Gly Gln Gly Thr Leu Val
                      100 105 110
          Thr Val Ser Ser
                  115
           <![CDATA[ <210> 112]]>
           <![CDATA[ <211> 107]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 light chain variable region]]>
           <![CDATA[ <400> 112]]>
          Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Gly Ala Ser Glu Asn Ile Tyr Gly Ala
                      20 25 30
          Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Lys Leu Leu Ile
                  35 40 45
          Tyr Gly Ala Thr Asn Leu Glu Asp Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Arg Asp Tyr Thr Phe Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Ile Ala Thr Tyr Tyr Cys Gln Asn Val Leu Thr Thr Pro Tyr
                          85 90 95
          Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100 105
           <![CDATA[ <210> 113]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 VH CDR1]]>
           <![CDATA[ <400> 113]]>
          Asp Tyr Asn Val Asn
          1 5
           <![CDATA[ <210> 114]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 VH CDR2]]>
           <![CDATA[ <400> 114]]>
          Val Ile Asn Pro Lys Tyr Gly Thr Thr Arg Tyr Asn Gln Lys Phe Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 115]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 VH CDR3]]>
           <![CDATA[ <400> 115]]>
          Gly Leu Asn Ala Trp Asp Tyr
          1 5
           <![CDATA[ <210> 116]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 VL CDR1]]>
           <![CDATA[ <400> 116]]>
          Gly Ala Ser Glu Asn Ile Tyr Gly Ala Leu Asn
          1 5 10
           <![CDATA[ <210> 117]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 VL CDR2]]>
           <![CDATA[ <400> 117]]>
          Gly Ala Thr Asn Leu Glu Asp
          1 5
           <![CDATA[ <210> 118]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h2A2 VL CDR3]]>
           <![CDATA[ <400> 118]]>
          Gln Asn Val Leu Thr Thr Pro Tyr Thr
          1 5
           <![CDATA[ <210> 119]]>
           <![CDATA[ <211> 117]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 heavy chain variable region]]>
           <![CDATA[ <400> 119]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Ser Gly Tyr
                      20 25 30
          Phe Met Asn Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Leu Ile Asn Pro Tyr Asn Gly Asp Ser Phe Tyr Asn Gln Lys Phe
              50 55 60
          Lys Gly Arg Val Thr Met Thr Arg Gln Thr Ser Thr Ser Thr Val Tyr
          65 70 75 80
          Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Val Arg Gly Leu Arg Arg Asp Phe Asp Tyr Trp Gly Gln Gly Thr Leu
                      100 105 110
          Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 120]]>
           <![CDATA[ <211> 112]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 light chain variable region]]>
           <![CDATA[ <400> 120]]>
          Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
          1 5 10 15
          Gln Pro Ala Ser Ile Ser Cys Lys Ser Ser Gln Ser Leu Leu Asp Ser
                      20 25 30
          Asp Gly Lys Thr Tyr Leu Asn Trp Leu Phe Gln Arg Pro Gly Gln Ser
                  35 40 45
          Pro Arg Arg Leu Ile Tyr Leu Val Ser Glu Leu Asp Ser Gly Val Pro
              50 55 60
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
          65 70 75 80
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Trp Gln Gly
                          85 90 95
          Thr His Phe Pro Arg Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 121]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 VH CDR1]]>
           <![CDATA[ <400> 121]]>
          Gly Tyr Phe Met Asn
          1 5
           <![CDATA[ <210> 122]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 VH CDR2]]>
           <![CDATA[ <400> 122]]>
          Leu Ile Asn Pro Tyr Asn Gly Asp Ser Phe Tyr Asn Gln Lys Phe Lys
          1 5 10 15
          Gly
           <![CDATA[ <210> 123]]>
           <![CDATA[ <211> 8]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 VH CDR3]]>
           <![CDATA[ <400> 123]]>
          Gly Leu Arg Arg Asp Phe Asp Tyr
          1 5
           <![CDATA[ <210> 124]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 VL CDR1]]>
           <![CDATA[ <400> 124]]>
          Lys Ser Ser Gln Ser Leu Leu Asp Ser Asp Gly Lys Thr Tyr Leu Asn
          1 5 10 15
           <![CDATA[ <210> 125]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 VL CDR2]]>
           <![CDATA[ <400> 125]]>
          Leu Val Ser Glu Leu Asp Ser
          1 5
           <![CDATA[ <210> 126]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: h15H3 VL CDR3]]>
           <![CDATA[ <400> 126]]>
          Trp Gln Gly Thr His Phe Pro Arg Thr
          1 5
           <![CDATA[ <210> 127]]>
           <![CDATA[ <211> 119]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A heavy chain variable region]]>
           <![CDATA[ <400> 127]]>
          Gln Val Gln Leu Gln Glu Ser Gly Pro Gly Leu Val Lys Pro Ser Glu
          1 5 10 15
          Thr Leu Ser Leu Thr Cys Thr Val Ser Gly Asp Ser Ile Thr Ser Gly
                      20 25 30
          Tyr Trp Asn Trp Ile Arg Gln Pro Gly Lys Gly Leu Glu Tyr Ile
                  35 40 45
          Gly Tyr Ile Ser Asp Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys
              50 55 60
          Ser Arg Val Thr Ile Ser Arg Asp Thr Ser Lys Asn Gln Tyr Ser Leu
          65 70 75 80
          Lys Leu Ser Ser Val Thr Ala Ala Asp Thr Ala Val Tyr Tyr Cys Ala
                          85 90 95
          Arg Arg Thr Leu Ala Thr Tyr Tyr Ala Met Asp Tyr Trp Gly Gln Gly
                      100 105 110
          Thr Leu Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 128]]>
           <![CDATA[ <211> 112]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A light chain variable region]]>
           <![CDATA[ <400> 128]]>
          Asp Phe Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
          1 5 10 15
          Gln Pro Ala Ser Ile Ser Cys Arg Ala Ser Gln Ser Leu Val His Ser
                      20 25 30
          Asp Gly Asn Thr Tyr Leu His Trp Tyr Gln Gln Arg Pro Gly Gln Ser
                  35 40 45
          Pro Arg Leu Leu Ile Tyr Arg Val Ser Asn Arg Phe Ser Gly Val Pro
              50 55 60
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
          65 70 75 80
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Ser Gln Ser
                          85 90 95
          Thr His Val Pro Pro Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 129]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A VH CDR1]]>
           <![CDATA[ <400> 129]]>
          Ser Gly Tyr Trp Asn
          1 5
           <![CDATA[ <210> 130]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A VH CDR2]]>
           <![CDATA[ <400> 130]]>
          Tyr Ile Ser Asp Ser Gly Ile Thr Tyr Tyr Asn Pro Ser Leu Lys Ser
          1 5 10 15
           <![CDATA[ <210> 131]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A VH CDR3]]>
           <![CDATA[ <400> 131]]>
          Arg Thr Leu Ala Thr Tyr Tyr Ala Met Asp Tyr
          1 5 10
           <![CDATA[ <210> 132]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A VL CDR1]]>
           <![CDATA[ <400> 132]]>
          Arg Ala Ser Gln Ser Leu Val His Ser Asp Gly Asn Thr Tyr Leu His
          1 5 10 15
           <![CDATA[ <210> 133]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A VL CDR2]]>
           <![CDATA[ <400> 133]]>
          Arg Val Ser Asn Arg Phe Ser
          1 5
           <![CDATA[ <210> 134]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-CD228A VL CDR3]]>
           <![CDATA[ <400> 134]]>
          Ser Gln Ser Thr His Val Pro Pro Thr
          1 5
           <![CDATA[ <210> 135]]>
           <![CDATA[ <211> 120]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A latituzumab vedotin (LV) heavy chain variable region]]>
           <![CDATA[ <400> 135]]>
          Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala
          1 5 10 15
          Ser Val Lys Val Ser Cys Lys Ala Ser Gly Leu Thr Ile Glu Asp Tyr
                      20 25 30
          Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met
                  35 40 45
          Gly Trp Ile Asp Pro Glu Asn Gly Asp Thr Glu Tyr Gly Pro Lys Phe
              50 55 60
          Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Asn Thr Ala Tyr
          65 70 75 80
          Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Val His Asn Ala His Tyr Gly Thr Trp Phe Ala Tyr Trp Gly Gln
                      100 105 110
          Gly Thr Leu Val Thr Val Ser Ser
                  115 120
           <![CDATA[ <210> 136]]>
           <![CDATA[ <211> 112]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A latituzumab vedotin (LV) light chain variable region]]>
           <![CDATA[ <400> 136]]>
          Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly
          1 5 10 15
          Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Leu His Ser
                      20 25 30
          Ser Gly Asn Thr Tyr Leu Glu Trp Tyr Gln Gln Arg Pro Gly Gln Ser
                  35 40 45
          Pro Arg Pro Leu Ile Tyr Lys Ile Ser Thr Arg Phe Ser Gly Val Pro
              50 55 60
          Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile
          65 70 75 80
          Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Tyr Cys Phe Gln Gly
                          85 90 95
          Ser His Val Pro Tyr Thr Phe Gly Gly Gly Thr Lys Val Glu Ile Lys
                      100 105 110
           <![CDATA[ <210> 137]]>
           <![CDATA[ <211> 5]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A latituzumab vedotin (LV) VH CDR1]]>
           <![CDATA[ <400> 137]]>
          Asp Tyr Tyr Met His
          1 5
           <![CDATA[ <210> 138]]>
           <![CDATA[ <211> 17]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A latituzumab vedotin (LV) VH CDR2]]>
           <![CDATA[ <400> 138]]>
          Trp Ile Asp Pro Glu Asn Gly Asp Thr Glu Tyr Gly Pro Lys Phe Gln
          1 5 10 15
          Gly
           <![CDATA[ <210> 139]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A latituzumab vedotin (LV) VH CDR3]]>
           <![CDATA[ <400> 139]]>
          His Asn Ala His Tyr Gly Thr Trp Phe Ala Tyr
          1 5 10
           <![CDATA[ <210> 140]]>
           <![CDATA[ <211> 16]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A Ladetuzumab Vidotine (LV) VL CDR1]]>
           <![CDATA[ <400> 140]]>
          Arg Ser Ser Gln Ser Leu Leu His Ser Ser Gly Asn Thr Tyr Leu Glu
          1 5 10 15
           <![CDATA[ <210> 141]]>
           <![CDATA[ <211> 7]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A Ladetuzumab Vidotine (LV) VL CDR2]]>
           <![CDATA[ <400> 141]]>
          Lys Ile Ser Thr Arg Phe Ser
          1 5
           <![CDATA[ <210> 142]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: SGN-LIV1A Ladetuzumab Vidotine (LV) VL CDR3]]>
           <![CDATA[ <400> 142]]>
          Phe Gln Gly Ser His Val Pro Tyr Thr
          1 5
           <![CDATA[ <210> 143]]>
           <![CDATA[ <211> 118]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) heavy chain variable region]]>
           <![CDATA[ <400> 143]]>
          Glu Val Gln Leu Leu Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
          1 5 10 15
          Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Asn Tyr
                      20 25 30
          Ala Met Ser Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
                  35 40 45
          Ser Ser Ile Ser Gly Ser Gly Asp Tyr Thr Tyr Tyr Thr Asp Ser Val
              50 55 60
          Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ser Lys Asn Thr Leu Tyr
          65 70 75 80
          Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys
                          85 90 95
          Ala Arg Ser Pro Trp Gly Tyr Tyr Leu Asp Ser Trp Gly Gln Gly Thr
                      100 105 110
          Leu Val Thr Val Ser Ser
                  115
           <![CDATA[ <210> 144]]>
           <![CDATA[ <211> 107]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) light chain variable region]]>
           <![CDATA[ <400> 144]]>
          Asp Ile Gln Met Thr Gln Ser Pro Pro Ser Leu Ser Ala Ser Ala Gly
          1 5 10 15
          Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Ser Ser Arg
                      20 25 30
          Leu Ala Trp Tyr Gln Gln Lys Pro Glu Lys Ala Pro Lys Ser Leu Ile
                  35 40 45
          Tyr Ala Ala Ser Ser Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly
              50 55 60
          Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro
          65 70 75 80
          Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Tyr Asn Ser Tyr Pro Tyr
                          85 90 95
          Thr Phe Gly Gln Gly Thr Lys Leu Glu Ile Lys
                      100 105
           <![CDATA[ <210> 145]]>
           <![CDATA[ <211> 8]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) VH CDR1]]>
           <![CDATA[ <400> 145]]>
          Gly Phe Thr Phe Ser Asn Tyr Ala
          1 5
           <![CDATA[ <210> 146]]>
           <![CDATA[ <211> 8]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) VH CDR2]]>
           <![CDATA[ <400> 146]]>
          Ile Ser Gly Ser Gly Asp Tyr Thr
          1 5
           <![CDATA[ <210> 147]]>
           <![CDATA[ <211> 11]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotumumab vedotin (TV) VH CDR3]]>
           <![CDATA[ <400> 147]]>
          Ala Arg Ser Pro Trp Gly Tyr Tyr Leu Asp Ser
          1 5 10
           <![CDATA[ <210> 148]]>
           <![CDATA[ <211> 6]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) VL CDR1]]>
           <![CDATA[ <400> 148]]>
          Gln Gly Ile Ser Ser Arg
          1 5
           <![CDATA[ <210> 149]]>
           <![CDATA[ <211> 3]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) VL CDR2]]>
           <![CDATA[ <400> 149]]>
          Ala Ala Ser
          1           
           <![CDATA[ <210> 150]]>
           <![CDATA[ <211> 9]]>
           <![CDATA[ <212> PRT]]>
           <![CDATA[ <213> Artificial sequences]]>
           <![CDATA[ <220>]]>
           <![CDATA[ <223> Synthesis: tesotuzumab vedotin (TV) VL CDR3]]>
           <![CDATA[ <400> 150]]>
          Gln Gln Tyr Asn Ser Tyr Pro Tyr Thr
          1 5

Figure 12_A0101_SEQ_0001
Figure 12_A0101_SEQ_0001

Figure 12_A0101_SEQ_0002
Figure 12_A0101_SEQ_0002

Figure 12_A0101_SEQ_0003
Figure 12_A0101_SEQ_0003

Figure 12_A0101_SEQ_0004
Figure 12_A0101_SEQ_0004

Figure 12_A0101_SEQ_0005
Figure 12_A0101_SEQ_0005

Figure 12_A0101_SEQ_0006
Figure 12_A0101_SEQ_0006

Figure 12_A0101_SEQ_0007
Figure 12_A0101_SEQ_0007

Figure 12_A0101_SEQ_0008
Figure 12_A0101_SEQ_0008

Figure 12_A0101_SEQ_0009
Figure 12_A0101_SEQ_0009

Figure 12_A0101_SEQ_0010
Figure 12_A0101_SEQ_0010

Figure 12_A0101_SEQ_0011
Figure 12_A0101_SEQ_0011

Figure 12_A0101_SEQ_0012
Figure 12_A0101_SEQ_0012

Figure 12_A0101_SEQ_0013
Figure 12_A0101_SEQ_0013

Figure 12_A0101_SEQ_0014
Figure 12_A0101_SEQ_0014

Figure 12_A0101_SEQ_0015
Figure 12_A0101_SEQ_0015

Figure 12_A0101_SEQ_0016
Figure 12_A0101_SEQ_0016

Figure 12_A0101_SEQ_0017
Figure 12_A0101_SEQ_0017

Figure 12_A0101_SEQ_0018
Figure 12_A0101_SEQ_0018

Figure 12_A0101_SEQ_0019
Figure 12_A0101_SEQ_0019

Figure 12_A0101_SEQ_0020
Figure 12_A0101_SEQ_0020

Figure 12_A0101_SEQ_0021
Figure 12_A0101_SEQ_0021

Figure 12_A0101_SEQ_0022
Figure 12_A0101_SEQ_0022

Figure 12_A0101_SEQ_0023
Figure 12_A0101_SEQ_0023

Figure 12_A0101_SEQ_0024
Figure 12_A0101_SEQ_0024

Figure 12_A0101_SEQ_0025
Figure 12_A0101_SEQ_0025

Figure 12_A0101_SEQ_0026
Figure 12_A0101_SEQ_0026

Figure 12_A0101_SEQ_0027
Figure 12_A0101_SEQ_0027

Figure 12_A0101_SEQ_0028
Figure 12_A0101_SEQ_0028

Figure 12_A0101_SEQ_0029
Figure 12_A0101_SEQ_0029

Figure 12_A0101_SEQ_0030
Figure 12_A0101_SEQ_0030

Figure 12_A0101_SEQ_0031
Figure 12_A0101_SEQ_0031

Figure 12_A0101_SEQ_0032
Figure 12_A0101_SEQ_0032

Figure 12_A0101_SEQ_0033
Figure 12_A0101_SEQ_0033

Figure 12_A0101_SEQ_0034
Figure 12_A0101_SEQ_0034

Figure 12_A0101_SEQ_0035
Figure 12_A0101_SEQ_0035

Figure 12_A0101_SEQ_0036
Figure 12_A0101_SEQ_0036

Figure 12_A0101_SEQ_0037
Figure 12_A0101_SEQ_0037

Figure 12_A0101_SEQ_0038
Figure 12_A0101_SEQ_0038

Figure 12_A0101_SEQ_0039
Figure 12_A0101_SEQ_0039

Figure 12_A0101_SEQ_0040
Figure 12_A0101_SEQ_0040

Figure 12_A0101_SEQ_0041
Figure 12_A0101_SEQ_0041

Figure 12_A0101_SEQ_0042
Figure 12_A0101_SEQ_0042

Figure 12_A0101_SEQ_0043
Figure 12_A0101_SEQ_0043

Figure 12_A0101_SEQ_0044
Figure 12_A0101_SEQ_0044

Figure 12_A0101_SEQ_0045
Figure 12_A0101_SEQ_0045

Figure 12_A0101_SEQ_0046
Figure 12_A0101_SEQ_0046

Figure 12_A0101_SEQ_0047
Figure 12_A0101_SEQ_0047

Figure 12_A0101_SEQ_0048
Figure 12_A0101_SEQ_0048

Figure 12_A0101_SEQ_0049
Figure 12_A0101_SEQ_0049

Figure 12_A0101_SEQ_0050
Figure 12_A0101_SEQ_0050

Figure 12_A0101_SEQ_0051
Figure 12_A0101_SEQ_0051

Figure 12_A0101_SEQ_0052
Figure 12_A0101_SEQ_0052

Figure 12_A0101_SEQ_0053
Figure 12_A0101_SEQ_0053

Figure 12_A0101_SEQ_0054
Figure 12_A0101_SEQ_0054

Figure 12_A0101_SEQ_0055
Figure 12_A0101_SEQ_0055

Figure 12_A0101_SEQ_0056
Figure 12_A0101_SEQ_0056

Figure 12_A0101_SEQ_0057
Figure 12_A0101_SEQ_0057

Figure 12_A0101_SEQ_0058
Figure 12_A0101_SEQ_0058

Figure 12_A0101_SEQ_0059
Figure 12_A0101_SEQ_0059

Figure 12_A0101_SEQ_0060
Figure 12_A0101_SEQ_0060

Figure 12_A0101_SEQ_0061
Figure 12_A0101_SEQ_0061

Figure 12_A0101_SEQ_0062
Figure 12_A0101_SEQ_0062

Figure 12_A0101_SEQ_0063
Figure 12_A0101_SEQ_0063

Figure 12_A0101_SEQ_0064
Figure 12_A0101_SEQ_0064

Figure 12_A0101_SEQ_0065
Figure 12_A0101_SEQ_0065

Figure 12_A0101_SEQ_0066
Figure 12_A0101_SEQ_0066

Figure 12_A0101_SEQ_0067
Figure 12_A0101_SEQ_0067

Figure 12_A0101_SEQ_0068
Figure 12_A0101_SEQ_0068

Figure 12_A0101_SEQ_0069
Figure 12_A0101_SEQ_0069

Figure 12_A0101_SEQ_0070
Figure 12_A0101_SEQ_0070

Figure 12_A0101_SEQ_0071
Figure 12_A0101_SEQ_0071

Figure 12_A0101_SEQ_0072
Figure 12_A0101_SEQ_0072

Figure 12_A0101_SEQ_0073
Figure 12_A0101_SEQ_0073

Figure 12_A0101_SEQ_0074
Figure 12_A0101_SEQ_0074

Figure 12_A0101_SEQ_0075
Figure 12_A0101_SEQ_0075

Figure 12_A0101_SEQ_0076
Figure 12_A0101_SEQ_0076

Figure 12_A0101_SEQ_0077
Figure 12_A0101_SEQ_0077

Figure 12_A0101_SEQ_0078
Figure 12_A0101_SEQ_0078

Figure 12_A0101_SEQ_0079
Figure 12_A0101_SEQ_0079

Figure 12_A0101_SEQ_0080
Figure 12_A0101_SEQ_0080

Figure 12_A0101_SEQ_0081
Figure 12_A0101_SEQ_0081

Figure 12_A0101_SEQ_0082
Figure 12_A0101_SEQ_0082

Figure 12_A0101_SEQ_0083
Figure 12_A0101_SEQ_0083

Figure 12_A0101_SEQ_0084
Figure 12_A0101_SEQ_0084

Figure 12_A0101_SEQ_0085
Figure 12_A0101_SEQ_0085

Figure 12_A0101_SEQ_0086
Figure 12_A0101_SEQ_0086

Figure 12_A0101_SEQ_0087
Figure 12_A0101_SEQ_0087

Figure 12_A0101_SEQ_0088
Figure 12_A0101_SEQ_0088

Claims (115)

一種治療癌症之方法，其包含向患有癌症之個體投與：(1)抗體-藥物結合物(ADC)，其包含結合腫瘤相關抗原之第一抗體及細胞毒性劑，其中該細胞毒性劑為微管蛋白干擾劑；及(2)第二抗體，其結合於免疫細胞接合子，其中該第二抗體包含增強與一或多種活化FcγR之結合的Fc，其中該等活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。A method of treating cancer, comprising administering to an individual suffering from cancer: (1) an antibody-drug conjugate (ADC) comprising a first antibody that binds a tumor-associated antigen and a cytotoxic agent, wherein the cytotoxic agent is A tubulin interfering agent; and (2) a second antibody that binds to an immune cell adaptor, wherein the second antibody comprises an Fc that enhances binding to one or more activated FcγRs, wherein the activated FcγRs include FcγRIIIa, FcγRIIa, and /or one or more of FcyRI. 如請求項1之方法，其中該第二抗體包含增強與至少FcγRIIIa之結合之Fc。The method of claim 1, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa. 如請求項1之方法，其中第二抗體包含增強與至少FcγRIIIa及FcγRIIa之結合的Fc。The method of claim 1, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRIIa. 如請求項1之方法，其中該第二抗體包含增強與至少FcγRIIIa及FcγRI之結合的Fc。The method of claim 1, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRI. 如請求項1之方法，其中該第二抗體包含增強與FcγRIIIa、FcγRIIa及FcγRI之結合的Fc。The method of claim 1, wherein the second antibody comprises an Fc that enhances binding to FcyRIIIa, FcyRIIa, and FcyRI. 如請求項1至5中任一項之方法，其中該第二抗體之該Fc減弱與一或多種抑制性FcγR之結合。The method of any one of claims 1 to 5, wherein the Fc of the second antibody attenuates binding to one or more inhibitory FcγRs. 如請求項6之方法，其中該第二抗體之該Fc減弱與FcγRIIb之結合。The method of claim 6, wherein the Fc of the second antibody attenuates binding to FcyRIIb. 如請求項1至7中任一項之方法，其中該第二抗體之該Fc具有減少之岩藻糖含量及/或已經工程改造以包含一或多個突變，使得該Fc增強與該一或多種活化FcγR之結合。The method of any one of claims 1 to 7, wherein the Fc of the second antibody has reduced fucose content and/or has been engineered to contain one or more mutations such that the Fc enhancement is associated with the one or Binding of various activated FcγRs. 如請求項8之方法，其中該第二抗體未經岩藻糖基化。The method of claim 8, wherein the second antibody is not fucosylated. 如請求項8之方法，其中該第二抗體包含重鏈恆定區中之取代S293D、A330L及I332E。The method of claim 8, wherein the second antibody comprises the substitutions S293D, A330L and I332E in the heavy chain constant region. 一種治療癌症之方法，其包含向患有癌症之個體投與抗體-藥物結合物，其中該抗體-藥物結合物包含：第一抗體，其與細胞毒性劑結合，其中該細胞毒性劑為微管蛋白干擾劑；及第二抗體，其結合免疫細胞接合子，其中該第二抗體未經岩藻糖基化。A method of treating cancer, comprising administering to an individual suffering from cancer an antibody-drug conjugate, wherein the antibody-drug conjugate comprises: a first antibody bound to a cytotoxic agent, wherein the cytotoxic agent is a microtubule a protein interfering agent; and a second antibody that binds to an immune cell engager, wherein the second antibody is not fucosylated. 如請求項1至11中任一項之方法，其中該第一抗體結合腫瘤相關抗原。The method of any one of claims 1 to 11, wherein the first antibody binds to a tumor-associated antigen. 一種治療癌症之方法，其包含向患有癌症之個體投與：(1)抗體-藥物結合物(ADC)，其中該ADC包含結合腫瘤相關抗原之第一抗體及細胞毒性劑，其中該細胞毒性劑為微管蛋白干擾劑；及(2)第二抗體，其結合免疫細胞接合子，其中該第二抗體包含相對於相同同種型之對應野生型Fc具有增強ADCC活性之Fc。A method of treating cancer, comprising administering to an individual suffering from cancer: (1) an antibody-drug conjugate (ADC), wherein the ADC comprises a primary antibody that binds a tumor-associated antigen and a cytotoxic agent, wherein the cytotoxic The agent is a tubulin interfering agent; and (2) a second antibody that binds an immune cell engager, wherein the second antibody comprises an Fc having enhanced ADCC activity relative to a corresponding wild-type Fc of the same isotype. 如請求項13之方法，其中該第二抗體包含相對於相同同種型之對應野生型Fc具有增強ADCC及ADCP活性之Fc。The method of claim 13, wherein the second antibody comprises an Fc having enhanced ADCC and ADCP activity relative to a corresponding wild-type Fc of the same isotype. 如請求項13或14之方法，其中該第二抗體未經岩藻糖基化。The method of claim 13 or 14, wherein the second antibody is not fucosylated. 如請求項13至15中任一項之方法，其中該第二抗體包含增強與一或多種活化FcγR之結合的Fc，其中該等活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。The method of any one of claims 13 to 15, wherein the second antibody comprises an Fc that enhances binding to one or more activated FcyRs, wherein the activated FcyRs comprise one or more of FcyRIIIa, FcyRIIa, and/or FcyRI . 如請求項16之方法，其中該第二抗體包含增強與至少FcγRIIIa之結合的Fc。The method of claim 16, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa. 如請求項16之方法，其中第二抗體包含增強與至少FcγRIIIa及FcγRIIa之結合的Fc。The method of claim 16, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRIIa. 如請求項16之方法，其中該第二抗體包含增強與至少FcγRIIIa及FcγRI之結合的Fc。The method of claim 16, wherein the second antibody comprises an Fc that enhances binding to at least FcyRIIIa and FcyRI. 如請求項16之方法，其中該第二抗體包含增強與FcγRIIIa、FcγRIIa及FcγRI之結合的Fc。The method of claim 16, wherein the second antibody comprises an Fc that enhances binding to FcyRIIIa, FcyRIIa, and FcyRI. 如請求項13至20中任一項之方法，其中該第二抗體之該Fc減弱與一或多種抑制性FcγR之結合。The method of any one of claims 13 to 20, wherein the Fc of the second antibody attenuates binding to one or more inhibitory FcγRs. 如請求項21之方法，其中該第二抗體之該Fc減弱與FcγRIIb之結合。The method of claim 21, wherein the Fc of the second antibody attenuates binding to FcyRIIb. 如請求項1至22中任一項之方法，其中該第一抗體結合選自以下之抗原：5T4 (TPBG)、ADAM-9、AG-7、ALK、ALP、AMHRII、APLP2、ASCT2、AVB6、AXL (UFO)、B7-H3 (CD276)、B7-H4、BCMA、C3a、C3b、C4.4a (LYPD3)、C5、C5a、CA6、CA9、CanAg、碳酸酐酶IX (CAIX)、組織蛋白酶D、CCR7、CD1、CD10、CD100、CD101、CD102、CD103、CD104、CD105、CD106、CD107a、CD107b、CD108、CD109、CD111、CD112、CD113、CD116、CD117、CD118、CD119、CD11A、CD11b、CD11c、CD120a、CD121a、CD121b、CD122、CD123、CD124、CD125、CD126、CD127、CD13、CD130、CD131、CD132、CD133、CD135、CD136、CD137、CD138、CD14、CD140a、CD140b、CD141、CD142、CD143、CD144、CD146、CD147、CD148、CD15、CD150、CD151、CD154、CD155、CD156a、CD156b、CD156c、CD157、CD158b2、CD158e、CD158f1、CD158h、CD158i、CD159a、CD16、CD160、CD161、CD162、CD163、CD164、CD166、CD167b、CD169、CD16a、CD16b、CD170、CD171、CD172a、CD172b、CD172g、CD18、CD180、CD181、CD183、CD184、CD185、CD19、CD194、CD197、CD1a、CD1b、CD1c、CD1d、CD2、CD20、CD200、CD201、CD202b、CD203c、CD204、CD205、CD206、CD208、CD21、CD213a1、CD213a2、CD217、CD218a、CD22、CD220、CD221、CD222、CD224、CD226、CD228、CD229、CD23、CD230、CD232、CD239、CD243、CD244、CD248、CD249、CD25、CD26、CD265、CD267、CD269、CD27、CD272、CD273、CD274、CD275、CD279、CD28、CD280、CD281、CD282、CD283、CD284、CD289、CD29、CD294、CD295、CD298、CD3、CD3 ε、CD30、CD300f、CD302、CD304、CD305、CD307、CD31、CD312、CD315、CD316、CD317、CD318、CD319、CD32、CD321、CD322、CD324、CD325、CD326、CD327、CD328、CD32b、CD33、CD331、CD332、CD333、CD334、CD337、CD339、CD34、CD340、CD344、CD35、CD352、CD36、CD37、CD38、CD39、CD3d、CD3g、CD4、CD41、CD42d、CD44、CD44v6、CD45、CD46、CD47、CD48、CD49a、CD49b、CD49c、CD49d、CD49e、CD49f、CD5、CD50、CD51、CD51 (整合素α-V)、CD52、CD53、CD54、CD55、CD56、CD58、CD59、CD6、CD61、CD62L、CD62P、CD63、CD64、CD66a-e、CD67、CD68、CD69、CD7、CD70、CD70L、CD71、CD71 (TfR)、CD72、CD73、CD74、CD79a、CD79b、CD8、CD80、CD82、CD83、CD84、CD85f、CD85i、CD85j、CD86、CD87、CD89、CD90、CD91、CD92、CD95、CD96、CD97、CD98、CDH6、CDH6 (鈣黏素6)、CDw210a、CDw210b、CEA、CEACAM5、CEACAM6、CFC1B、cKIT、CLDN18.2 (密連蛋白18.2)、CLDN6、CLDN9、CLL-1、c-MET、補體因子C3、Cripto、CSP-1、CXCR5、DCLK1、DLK-1、DLL3、DPEP3、DR5 (死亡受體5)、抗黏附素(Dysadherin)、EFNA4、EGFR、EGFR野生型、EGFRviii、EGP-1 (TROP-2)、EGP-2、EMP2、ENPP3、EpCAM、EphA2、EphA3、Ephrin-A4 (EFNA4)、ETBR、FAP、FcRH5、FGFR2、FGFR3、FLT3、FOLR、FOLR1、FOLR-α、FSH、GCC、GD2、GD3、globo H、GPC1、GPC-1、GPC3、GPNMB、GPR20、HER2、HER-2、HER3、HER-3、HGFR (c-Met)、HLA-DR、HM1.24、HSP90、Ia、IGF-1R、IL-13R、IL-15、IL1RAP、IL-2、IL-3、IL-4、IL7R、整合素αVβ3(integrin alphaVbeta3/integrin αVβ3)、整合素β-6、介白素-4受體(IL4R)、KAAG-1、KLK2、LAMP-1、Le(y)、路易斯Y抗原(Lewis Y antigen)、LGALS3BP、LGR5、LH/hCG、LHRH、脂筏、LIV-1 (SLC39A6或ZIP6)、LRP-1、LRRC15、LY6E、巨噬細胞甘露糖受體1、MAGE、間皮素(MSLN)、MET、I類MHC鏈相關蛋白A及B (MICA及MICB)、MN/CA IX、MRC2、MT1-MMP、MTX3、MTX5、MUC1、MUC16、MUC2、MUC3、MUC4、MUC5、MUC5ac、NaPi2b、NCA-90、NCA-95、連接素-4、Notch3、核仁素、OAcGD2、OT-MUC1 (腫瘤栓繫MUC1)、OX001L、P1GF、PAM4抗原、p-鈣黏素(鈣黏素3)、PD-L1、磷脂醯基絲胺酸(PS)、PRLR、促乳素受體(PRLR)、假單胞菌(Pseudomonas)、PSMA、PTK4、PTK7、受體酪胺酸激酶(RTK)、RNF43、ROR1、ROR2、SAIL、SEZ6、SLAMF7、SLC44A4、SLITRK6、SLMAMF7 (CS1)、SLTRK6、分選蛋白(Sortilin) (SORT1)、SSEA-4、SSTR2、金黃色葡萄球菌(Staphylococcus aureus) (抗生素劑)、STEAP-1、STING、STn、T101、TAA、TAC、TDGF1、肌腱蛋白、TENB2、TGF-B、Thomson-Friedenreich抗原、Thy1.1、TIM-1、組織因子(TF；CD142)、TM4SF1、Tn抗原、TNF-alpha (TNFα)、TRA-1-60、TRAIL受體(R1及R2)、TROP-2、腫瘤相關糖蛋白72 (TAG-72)、uPAR、VEGFR、VEGFR-2及xCT。The method of any one of claims 1 to 22, wherein the first antibody binds an antigen selected from the group consisting of: 5T4 (TPBG), ADAM-9, AG-7, ALK, ALP, AMHRII, APLP2, ASCT2, AVB6, AXL (UFO), B7-H3 (CD276), B7-H4, BCMA, C3a, C3b, C4.4a (LYPD3), C5, C5a, CA6, CA9, CanAg, Carbonic Anhydrase IX (CAIX), Cathepsin D , CCR7, CD1, CD10, CD100, CD101, CD102, CD103, CD104, CD105, CD106, CD107a, CD107b, CD108, CD109, CD111, CD112, CD113, CD116, CD117, CD118, CD119, CD11A, CD11b, CD11c, CD120a , CD121a, CD121b, CD122, CD123, CD124, CD125, CD126, CD127, CD13, CD130, CD131, CD132, CD133, CD135, CD136, CD137, CD138, CD14, CD140a, CD140b, CD141, CD142, CD143, CD144, CD146 , CD147, CD148, CD15, CD150, CD151, CD154, CD155, CD156a, CD156b, CD156c, CD157, CD158b2, CD158e, CD158f1, CD158h, CD158i, CD159a, CD16, CD160, CD161, CD162, CD163, CD6664, CD163 , CD169, CD16a, CD16b, CD170, CD171, CD172a, CD172b, CD172g, CD18, CD180, CD181, CD183, CD184, CD185, CD19, CD194, CD197, CD1a, CD1b, CD1c, CD1d, CD2, CD20, CD200, CD201 , CD202b, CD203c, CD204, CD205, CD206, CD208, CD21, CD213a1, CD213a2, CD217, CD218a, CD22, CD220, CD221, CD222, CD224, CD226, CD228, CD229, CD23, CD230, CD232, CD239, CD243, CD239 , CD248, CD249, CD25, CD26, CD265, CD267, CD269, CD27, CD272, CD2 73, CD274, CD275, CD279, CD28, CD280, CD281, CD282, CD283, CD284, CD289, CD29, CD294, CD295, CD298, CD3, CD3ε, CD30, CD300f, CD302, CD304, CD305, CD307, CD31, CD312 , CD315, CD316, CD317, CD318, CD319, CD32, CD321, CD322, CD324, CD325, CD326, CD327, CD328, CD32b, CD33, CD331, CD332, CD333, CD334, CD337, CD339, CD34, CD340, CD344, CD35 , CD352, CD36, CD37, CD38, CD39, CD3d, CD3g, CD4, CD41, CD42d, CD44, CD44v6, CD45, CD46, CD47, CD48, CD49a, CD49b, CD49c, CD49d, CD49e, CD49f, CD5, CD50, CD51 , CD51 (integrin alpha-V), CD52, CD53, CD54, CD55, CD56, CD58, CD59, CD6, CD61, CD62L, CD62P, CD63, CD64, CD66a-e, CD67, CD68, CD69, CD7, CD70, CD70L, CD71, CD71 (TfR), CD72, CD73, CD74, CD79a, CD79b, CD8, CD80, CD82, CD83, CD84, CD85f, CD85i, CD85j, CD86, CD87, CD89, CD90, CD91, CD92, CD95, CD96 , CD97, CD98, CDH6, CDH6 (Cadherin 6), CDw210a, CDw210b, CEA, CEACAM5, CEACAM6, CFC1B, cKIT, CLDN18.2 (claudin 18.2), CLDN6, CLDN9, CLL-1, c-MET , complement factor C3, Cripto, CSP-1, CXCR5, DCLK1, DLK-1, DLL3, DPEP3, DR5 (death receptor 5), anti-adhesin (Dysadherin), EFNA4, EGFR, EGFR wild-type, EGFRviii, EGP- 1 (TROP-2), EGP-2, EMP2, ENPP3, EpCAM, EphA2, EphA3, Ephrin-A4 (EFNA4), ETBR, FAP, FcRH5, FGFR2, FGFR3, FLT3, FOLR, FOLR1, F OLR-α, FSH, GCC, GD2, GD3, globo H, GPC1, GPC-1, GPC3, GPNMB, GPR20, HER2, HER-2, HER3, HER-3, HGFR (c-Met), HLA-DR, HM1.24, HSP90, Ia, IGF-1R, IL-13R, IL-15, IL1RAP, IL-2, IL-3, IL-4, IL7R, integrin alphaVbeta3 (integrin alphaVbeta3/integrin alphaVbeta3), integrin beta -6. Interleukin-4 receptor (IL4R), KAAG-1, KLK2, LAMP-1, Le(y), Lewis Y antigen, LGALS3BP, LGR5, LH/hCG, LHRH, lipid rafts , LIV-1 (SLC39A6 or ZIP6), LRP-1, LRRC15, LY6E, macrophage mannose receptor 1, MAGE, mesothelin (MSLN), MET, MHC class I chain-associated proteins A and B (MICA and MICB), MN/CA IX, MRC2, MT1-MMP, MTX3, MTX5, MUC1, MUC16, MUC2, MUC3, MUC4, MUC5, MUC5ac, NaPi2b, NCA-90, NCA-95, connexin-4, Notch3, nuclear Renin, OAcGD2, OT-MUC1 (tumor tethered MUC1), OX001L, P1GF, PAM4 antigen, p-cadherin (cadherin 3), PD-L1, phosphatidylserine (PS), PRLR, Prolactin receptor (PRLR), Pseudomonas, PSMA, PTK4, PTK7, receptor tyrosine kinase (RTK), RNF43, ROR1, ROR2, SAIL, SEZ6, SLAMF7, SLC44A4, SLITRK6, SLMAMF7 (CS1), SLTRK6, Sortilin (SORT1), SSEA-4, SSTR2, Staphylococcus aureus (antibiotic agent), STEAP-1, STING, STn, T101, TAA, TAC, TDGF1 , Tenascin, TENB2, TGF-B, Thomson-Friedenreich antigen, Thy1.1, TIM-1, tissue factor (TF; CD142), TM4SF1, Tn antigen, TNF-alpha (TNFα), TRA-1-60, TRAIL Receptors (R1 and R2), TROP-2, Tumor-Associated Glycoprotein 72 (TAG-72), uPAR, VEGFR, VEGFR-2 and xCT. 如請求項1至23中任一項之方法，其中該第一抗體不結合連接素-4。The method of any one of claims 1 to 23, wherein the first antibody does not bind connexin-4. 如請求項1至24中任一項之方法，其中該方法不包含投與抗體-藥物結合物，該抗體-藥物結合物包含結合連接素-4之抗體。The method of any one of claims 1 to 24, wherein the method does not comprise administering an antibody-drug conjugate comprising an antibody that binds connexin-4. 如請求項1至25中任一項之方法，其中該第一抗體結合選自以下之抗原：CD71、Axl、AMHRII及LGR5、Axl、CA9、CD142、CD20、CD22、CD228、CD248、CD30、CD33、CD37、CD48、CD7、CD71、CD79b、CLDN18.2、CLDN6、c-MET、EGFR、EphA2、ETBR、FCRH5、GCC、Globo H、gpNMB、HER-2、IL7R、整合素β-6、KAAG-1、LGR5、LIV-1、LRRC15、Ly6E、間皮素(MSLN)、MET、MRC2、MUC16、NaPi2b、連接素-4、OT-MUC1 (腫瘤栓繫-MUC1)、PSMA、ROR1、SLAMF7、SLC44A4、SLITRK6、STEAP-1、STn、TIM-1、TRA-1-60及腫瘤相關糖蛋白72 (TAG-72)。The method of any one of claims 1 to 25, wherein the first antibody binds an antigen selected from the group consisting of CD71, Axl, AMHRII and LGR5, Axl, CA9, CD142, CD20, CD22, CD228, CD248, CD30, CD33 , CD37, CD48, CD7, CD71, CD79b, CLDN18.2, CLDN6, c-MET, EGFR, EphA2, ETBR, FCRH5, GCC, Globo H, gpNMB, HER-2, IL7R, integrin beta-6, KAAG- 1. LGR5, LIV-1, LRRC15, Ly6E, mesothelin (MSLN), MET, MRC2, MUC16, NaPi2b, connexin-4, OT-MUC1 (tumor tethering-MUC1), PSMA, ROR1, SLAMF7, SLC44A4 , SLITRK6, STEAP-1, STn, TIM-1, TRA-1-60 and tumor-associated glycoprotein 72 (TAG-72). 如請求項1至25中任一項之方法，其中該第一抗體結合選自以下之抗原：BCMA、GPC1、CD30、cMET、SAIL、HER3、CD70、CD46、CD48、HER2、5T4、ENPP3、CD19、EGFR及EphA2。The method of any one of claims 1 to 25, wherein the first antibody binds an antigen selected from the group consisting of BCMA, GPC1, CD30, cMET, SAIL, HER3, CD70, CD46, CD48, HER2, 5T4, ENPP3, CD19 , EGFR and EphA2. 如請求項1至25中任一項之方法，其中該第一抗體結合選自以下之抗原：Her2、TROP2、BCMA、cMet、整合素αVβ6 (integrin alphVbeta6/integrin αVβ6)、CD22、CD79b、CD30、CD19、CD70、CD228、CD47及CD48。The method of any one of claims 1 to 25, wherein the first antibody binds an antigen selected from the group consisting of Her2, TROP2, BCMA, cMet, integrin alphVbeta6/integrin αVβ6, CD22, CD79b, CD30, CD19, CD70, CD228, CD47 and CD48. 如請求項1至25中任一項之方法，其中該第一抗體結合選自以下之抗原：CD142、整合素β-6、整合素αVβ6、ENPP3、CD19、Ly6E、cMET、C4.4a、CD37、MUC16、STEAP-1、LRRC15、SLITRK6、ETBR、FCRH5、Axl、EGFR、CD79b、BCMA、CD70、PSMA、CD79b、CD228、CD48、LIV-1、EphA2、SLC44A4、CD30及sTn。The method of any one of claims 1 to 25, wherein the first antibody binds an antigen selected from the group consisting of CD142, integrin beta-6, integrin alphaVbeta6, ENPP3, CD19, Ly6E, cMET, C4.4a, CD37 , MUC16, STEAP-1, LRRC15, SLITRK6, ETBR, FCRH5, Axl, EGFR, CD79b, BCMA, CD70, PSMA, CD79b, CD228, CD48, LIV-1, EphA2, SLC44A4, CD30 and sTn. 如請求項1至29中任一項之方法，其中該微管蛋白干擾劑為奧瑞他汀(auristatins)、微管蛋白裂解素(tubulysin)、秋水仙鹼(colchicine)、長春花生物鹼(vinca alkaloid)、紫杉烷(taxane)、克瑞托欣(cryptophycin)、類美登素(maytansinoid)或哈米特林(hemiasterlin)。The method of any one of claims 1 to 29, wherein the tubulin interfering agent is auristatins, tubulysin, colchicine, vinca alkaloids alkaloid), taxane, cryptophycin, maytansinoid or hemiasterlin. 如請求項30之方法，其中該微管蛋白干擾劑為奧瑞他汀。The method of claim 30, wherein the tubulin interfering agent is auristatin. 如請求項1至31中任一項之方法，其中該微管蛋白干擾劑為尾海兔素(dolostatin)-10、MMAE (N-甲基纈胺酸-纈胺酸-多拉索因(dolaisoleuine)-多拉普因(dolaproine)-降麻黃鹼)、MMAF (N-甲基纈胺酸-纈胺酸-多拉索因-多拉普因-苯丙胺酸)、奧瑞他汀F、AEB、AEVB或AFP (奧瑞他汀苯丙胺酸苯二胺)。The method of any one of claims 1 to 31, wherein the tubulin interfering agent is dolostatin-10, MMAE (N-methylvaline-valine-dorasoine ( dolaisoleuine)-dolaproine-noephedrine), MMAF (N-methylvaline-valine-dorasoine-dorapine-phenylalanine), auristatin F, AEB, AEVB or AFP (auristatin phenylalanine phenylenediamine). 如請求項1至32中任一項之方法，其中該微管蛋白干擾劑為MMAE。The method of any one of claims 1 to 32, wherein the tubulin interfering agent is MMAE. 如請求項33之方法，其中該MMAE經由包含纈胺酸及瓜胺酸之連接子與該第一抗體結合。The method of claim 33, wherein the MMAE is bound to the first antibody via a linker comprising valine and citrulline. 如請求項34之方法，其中該連接子-MMAE為vcMMAE。The method of claim 34, wherein the linker-MMAE is vcMMAE. 如請求項33之方法，其中該MMAE經由包含白胺酸、丙胺酸及麩胺酸之連接子與該第一抗體結合。The method of claim 33, wherein the MMAE binds to the first antibody via a linker comprising leucine, alanine, and glutamic acid. 如請求項36之方法，其中該連接子-MMAE為dLAE-MMAE。The method of claim 36, wherein the linker-MMAE is dLAE-MMAE. 如請求項1至32中任一項之方法，其中該微管蛋白干擾劑為MMAF。The method of any one of claims 1 to 32, wherein the tubulin interfering agent is MMAF. 如請求項1至32中任一項之方法，其中該微管蛋白干擾劑為微管蛋白裂解素(tubulysin)。The method of any one of claims 1 to 32, wherein the tubulin interfering agent is tubulysin. 如請求項39之方法，其中該微管蛋白裂解素係選自微管蛋白裂解素D、微管蛋白裂解素M、微管苯丙胺酸及微管酪胺酸。The method of claim 39, wherein the tubulin lysin is selected from the group consisting of tubulin D, tubulin M, tubulin phenylalanine and tubulin tyrosine. 如請求項1至32中任一項之方法，其中該抗體-藥物結合物係選自AbGn-107 (Ab1-18Hr1)、AGS62P1 (ASP1235)、ALT-P7 (HM2-MMAE)、BA3011 (CAB-AXL-ADC)、貝蘭單抗莫福汀(belantamab mafodotin)、本妥昔單抗維多汀(brentuximab vedotin)、瑟吐珠單抗維多汀(cirmtuzumab vedotin) (VLS-101、UC-961ADC3)、考非妥珠單抗比利多汀(cofetuzumab pelidotin) (PF-06647020、PTK7-ADC、PF-7020、ABBV-647)、CX-2029 (ABBV-2029)、迪西妥單抗維多汀(disitamab vedotin) (RC48)、伊那波它單抗維多汀(enapotamab vedotin) (HuMax-AXL-ADC、AXL-107-MMAE)、恩諾單抗維多汀(enfortumab vedotin) (EV)、FS-1502 (LCB14-0110)、吉妥單抗奧佐米星(gemtuzumab ozogamicin)、HTI-1066 (SHR-A1403)、英妥珠單抗奧佐米星(inotuzumab ozogamicin)、PF-06804103 (NG-HER2 ADC)、波妥珠單抗維多汀(polatuzumab vedotin)、薩西土珠單抗戈維特坎(sacituzumab govitecan)、SGN-B6A、SGN-CD228A、SGN-STNV、STI-6129 (CD38 ADC、LNDS1001、CD38-077 ADC)、肽利索單抗維多汀(telisotuzumab vedotin) (ABBV-399)、替索圖單抗維多汀(tisotumab vedotin) (Humax-TF-ADC、tf-011-mmae、TV)、曲妥珠單抗德魯特坎(trastuzumab deruxtecan)、曲妥珠單抗恩他新(trastuzumab emtansine)及伏司妥珠單抗莫福汀(vorsetuzumab mafodotin)。The method of any one of claims 1 to 32, wherein the antibody-drug conjugate is selected from the group consisting of AbGn-107 (Ab1-18Hr1), AGS62P1 (ASP1235), ALT-P7 (HM2-MMAE), BA3011 (CAB- AXL-ADC), belantamab mafodotin, brentuximab vedotin, cirmtuzumab vedotin (VLS-101, UC-961ADC3 ), cofetuzumab pelidotin (PF-06647020, PTK7-ADC, PF-7020, ABBV-647), CX-2029 (ABBV-2029), dicituzumab vedotin (disitamab vedotin) (RC48), enapotamab vedotin (HuMax-AXL-ADC, AXL-107-MMAE), enfortumab vedotin (EV), FS -1502 (LCB14-0110), gemtuzumab ozogamicin, HTI-1066 (SHR-A1403), inotuzumab ozogamicin, PF-06804103 (NG- HER2 ADC), polatuzumab vedotin, sacituzumab govitecan, SGN-B6A, SGN-CD228A, SGN-STNV, STI-6129 (CD38 ADC, LNDS1001 , CD38-077 ADC), telisotuzumab vedotin (ABBV-399), tisotumab vedotin (Humax-TF-ADC, tf-011-mmae, TV ), trastuzumab deruxtecan, trastuzumab emtansine, and vorsetuzumab mafodotin. 如請求項1至41中任一項之方法，其中該第一抗體為抗密連蛋白-18.2抗體，其包含分別包含SEQ ID NO:61至66之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-claudin-18.2 antibody comprising heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3. 如請求項42之方法，其中該抗密連蛋白-18.2抗體包含：包含SEQ ID NO: 59之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO: 60之胺基酸序列的輕鏈可變區(VL)。The method of claim 42, wherein the anti-Claudin-18.2 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 59; and an amino acid comprising SEQ ID NO: 60 Sequence of the light chain variable region (VL). 如請求項43之方法，其中該抗密連蛋白-18.2抗體為唑貝妥西單抗(zolbetuximab) (175D10)。The method of claim 43, wherein the anti-claudin-18.2 antibody is zolbetuximab (175D10). 如請求項1至41中任一項之方法，其中該第一抗體為抗密連蛋白-18.2抗體，其包含分別包含SEQ ID NO:69至74之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-claudin-18.2 antibody comprising heavy chain CDR1, CDR2 and CDR2 comprising the amino acid sequences of SEQ ID NOs: 69 to 74, respectively CDR3 and light chain CDR1, CDR2 and CDR3. 如請求項45之方法，其中該抗密連蛋白-18.2抗體包含：包含SEQ ID NO: 67之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO: 68之胺基酸序列的輕鏈可變區(VL)。The method of claim 45, wherein the anti-Claudin-18.2 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 67; and an amino acid comprising SEQ ID NO: 68 Sequence of the light chain variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗PD-L1抗體，其包含分別包含SEQ ID NO:77至82之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-PD-L1 antibody comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 77 to 82, respectively, and Light chain CDR1, CDR2 and CDR3. 如請求項47之方法，其中該抗PD-L1抗體包含：包含SEQ ID NO:75之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:76之胺基酸序列的輕鏈可變區(VL)。The method of claim 47, wherein the anti-PD-L1 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:75; and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:76 Light chain variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗ALP抗體，其包含分別包含SEQ ID NO:85至90之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-ALP antibody comprising a heavy chain CDR1, CDR2 and CDR3 and a light chain comprising the amino acid sequences of SEQ ID NOs: 85 to 90, respectively CDR1, CDR2 and CDR3. 如請求項49之方法，其中該ALP抗體包含：包含SEQ ID NO:83之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:84之胺基酸序列的輕鏈可變區(VL)。The method of claim 49, wherein the ALP antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:83; and a light chain variable region comprising the amino acid sequence of SEQ ID NO:84 variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體包含抗B7H4抗體，其包含分別包含SEQ ID NO:93至98之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody comprises an anti-B7H4 antibody comprising heavy chain CDR1, CDR2 and CDR3 and a light chain comprising the amino acid sequences of SEQ ID NOs: 93 to 98, respectively CDR1, CDR2 and CDR3. 如請求項51之方法，其中該抗B7H4抗體包含：包含SEQ ID NO:91之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:92之胺基酸序列的輕鏈可變區(VL)。The method of claim 51, wherein the anti-B7H4 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:91; and a light chain comprising the amino acid sequence of SEQ ID NO:92 variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗HER2抗體，其包含：包含SEQ ID NO:99之胺基酸序列的重鏈及包含SEQ ID NO:100之胺基酸序列的輕鏈。The method of any one of claims 1 to 41, wherein the first antibody is an anti-HER2 antibody comprising: a heavy chain comprising the amino acid sequence of SEQ ID NO:99 and an amine group comprising SEQ ID NO:100 acid sequence of the light chain. 如請求項53之方法，其中該抗體-藥物結合物為迪西妥單抗維多汀。The method of claim 53, wherein the antibody-drug conjugate is dicituzumab vedotin. 如請求項1至41中任一項之方法，其中該第一抗體為抗NaPi2B抗體，其包含：包含SEQ ID NO:101之胺基酸序列的重鏈及包含SEQ ID NO:102之胺基酸序列的輕鏈。The method of any one of claims 1 to 41, wherein the first antibody is an anti-NaPi2B antibody comprising: a heavy chain comprising the amino acid sequence of SEQ ID NO: 101 and an amino group comprising SEQ ID NO: 102 acid sequence of the light chain. 如請求項55之方法，其中該抗體-藥物結合物為立伐土珠單抗維多汀(lifastuzumab vedotin)。The method of claim 55, wherein the antibody-drug conjugate is lifastuzumab vedotin. 如請求項1至41中任一項之方法，其中該第一抗體為抗連接素-4抗體，其包含分別包含SEQ ID NO:105至110之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-connexin-4 antibody comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 105 to 110, respectively and light chain CDR1, CDR2 and CDR3. 如請求項57之方法，其中該抗連接素-4抗體為包含以下之抗體：包含SEQ ID NO:103之胺基酸序列的重鏈可變區(VH)及包含SEQ ID NO:104之胺基酸序列的輕鏈可變區(VL)。The method of claim 57, wherein the anti-connexin-4 antibody is an antibody comprising a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:103 and an amine comprising SEQ ID NO:104 The light chain variable region (VL) of the amino acid sequence. 如請求項58之方法，其中該抗體-藥物結合物為恩諾單抗維多汀。The method of claim 58, wherein the antibody-drug conjugate is ennozumab vedotin. 如請求項1至41中任一項之方法，其中該第一抗體為抗AVB6抗體，其包含分別包含SEQ ID NO:113至118之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-AVB6 antibody comprising heavy chain CDR1, CDR2 and CDR3 and a light chain comprising the amino acid sequences of SEQ ID NOs: 113 to 118, respectively CDR1, CDR2 and CDR3. 如請求項60之方法，其中該抗AVB6抗體包含：包含SEQ ID NO:37之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:38之胺基酸序列的輕鏈可變區(VL)。The method of claim 60, wherein the anti-AVB6 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:37; and a light chain comprising the amino acid sequence of SEQ ID NO:38 variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗AVB6抗體，其包含分別包含SEQ ID NO:121至126之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-AVB6 antibody comprising a heavy chain CDR1, CDR2 and CDR3 and a light chain comprising the amino acid sequences of SEQ ID NOs: 121 to 126, respectively CDR1, CDR2 and CDR3. 如請求項62之方法，其中該抗AVB6抗體包含：包含SEQ ID NO:119之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:120之胺基酸序列的輕鏈可變區(VL)。The method of claim 62, wherein the anti-AVB6 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO: 119; and a light chain comprising the amino acid sequence of SEQ ID NO: 120 variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗CD228抗體，其包含分別包含SEQ ID NO:129至134之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-CD228 antibody comprising a heavy chain CDR1, CDR2 and CDR3 and a light chain comprising the amino acid sequences of SEQ ID NOs: 129 to 134, respectively CDR1, CDR2 and CDR3. 如請求項64之方法，其中該抗CD228抗體包含：包含SEQ ID NO:127之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:128之胺基酸序列的輕鏈可變區(VL)。The method of claim 64, wherein the anti-CD228 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:127; and a light chain comprising the amino acid sequence of SEQ ID NO:128 variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗LIV-1抗體，其包含分別包含SEQ ID NO:137至142之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-LIV-1 antibody comprising heavy chain CDR1, CDR2 and CDR3 comprising the amino acid sequences of SEQ ID NOs: 137 to 142, respectively, and Light chain CDR1, CDR2 and CDR3. 如請求項66之方法，其中該抗LIV-1抗體包含：包含SEQ ID NO:135之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:136之胺基酸序列的輕鏈可變區(VL)。The method of claim 66, wherein the anti-LIV-1 antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:135; and a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:136 Light chain variable region (VL). 如請求項1至41中任一項之方法，其中該第一抗體為抗組織因子抗體，其包含分別包含SEQ ID NO:145至150之胺基酸序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of any one of claims 1 to 41, wherein the first antibody is an anti-tissue factor antibody comprising heavy chain CDR1, CDR2 and CDR3 and a light chain comprising the amino acid sequences of SEQ ID NOs: 145 to 150, respectively Chains CDR1, CDR2 and CDR3. 如請求項68之方法，其中該抗組織因子抗體包含：包含SEQ ID NO:143之胺基酸序列的重鏈可變區(VH)；及包含SEQ ID NO:144之胺基酸序列的輕鏈可變區(VL)。The method of claim 68, wherein the anti-tissue factor antibody comprises: a heavy chain variable region (VH) comprising the amino acid sequence of SEQ ID NO:143; and a light chain comprising the amino acid sequence of SEQ ID NO:144 chain variable region (VL). 如請求項69之方法，其中該抗體-藥物結合物為替索圖單抗維多汀。The method of claim 69, wherein the antibody-drug conjugate is tesotuzumab vedotin. 如請求項1至70中任一項之方法，其中該第二抗體結合選自以下之免疫細胞接合子：抗苗勒氏激素受體(anti-Mullerian Hormone Receptor) II (AMHR2)、B7、B7H1、B7H2、B7H3、B7H4、BAFF-R、BCMA (B細胞成熟抗原)、Bst1/CD157、C5補體、CC趨化因子受體4 (CCR4)、CD123、CD137、CD19、CD20、CD25 (IL2RA)、CD276、CD278、CD3、CD32、CD33、CD37、CD38、CD4及HIV-1 gp120結合位點、CD40、CD70、CD70 (TNF受體配位體家族中之成員)、CD80、CD86、密連蛋白18.2、c-MET、CSF1R、CTLA-4、EGFR、EGFR MET原癌基因、EPHA3、ERBB2、ERBB3、FGFR2b、FLT3、GITR、糖皮質激素誘導之TNF受體(GITR)、HER2、HER3、HLA、ICOS、IDO1、IFNAR1、IFNAR2、IGF-1R、IL-3Rα (CD123)、IL-5R、IL-5Rα、LAG-3、MET原癌基因、OX40 (CD134)、PD-1、PD-L1、PD-L2、PVRIG、EBOV糖蛋白(GP)之呼吸道融合病毒(RSV)重糖基化黏蛋白樣域、恆河猴(Rh) D、唾液酸免疫球蛋白樣凝集素8 (Siglec-8)、傳信淋巴球活化分子(SLAMF7/CS1)、T細胞受體細胞毒性T淋巴球相關抗原4 (CTLA4)、TIGIT、TIM3 (HAVCR2)、Muc1之腫瘤特異性糖抗原決定基(glycoepitope) (TA-Muc1)、VSIR (VISTA)及VTCN1。The method of any one of claims 1 to 70, wherein the second antibody binds to an immune cell engager selected from the group consisting of: anti-Mullerian Hormone Receptor II (AMHR2), B7, B7H1 , B7H2, B7H3, B7H4, BAFF-R, BCMA (B cell maturation antigen), Bst1/CD157, C5 complement, CC chemokine receptor 4 (CCR4), CD123, CD137, CD19, CD20, CD25 (IL2RA), CD276, CD278, CD3, CD32, CD33, CD37, CD38, CD4 and HIV-1 gp120 binding site, CD40, CD70, CD70 (members of the TNF receptor ligand family), CD80, CD86, claudin 18.2 , c-MET, CSF1R, CTLA-4, EGFR, EGFR MET proto-oncogene, EPHA3, ERBB2, ERBB3, FGFR2b, FLT3, GITR, glucocorticoid-induced TNF receptor (GITR), HER2, HER3, HLA, ICOS , IDO1, IFNAR1, IFNAR2, IGF-1R, IL-3Rα (CD123), IL-5R, IL-5Rα, LAG-3, MET proto-oncogene, OX40 (CD134), PD-1, PD-L1, PD- L2, PVRIG, Respiratory fusion virus (RSV) heavily glycosylated mucin-like domain of EBOV glycoprotein (GP), Rhesus monkey (Rh) D, Sialyl immunoglobulin-like lectin 8 (Siglec-8), Signaling lymphocyte activation molecule (SLAMF7/CS1), T cell receptor cytotoxic T lymphocyte-associated antigen 4 (CTLA4), TIGIT, TIM3 (HAVCR2), tumor-specific glycoepitope of Muc1 (TA-Muc1) ), VSIR (VISTA), and VTCN1. 如請求項1至71中任一項之方法，其中該第二抗體結合TIGIT。The method of any one of claims 1 to 71, wherein the second antibody binds TIGIT. 如請求項72之方法，其中該第二抗體包含： (a)    重鏈CDR1，其包含選自SEQ ID NO:7至9之胺基酸序列； (b)    重鏈CDR2，其包含選自SEQ ID NO:10至13之胺基酸序列； (c)    重鏈CDR3，其包含選自SEQ ID NO:14至16之胺基酸序列； (d)    輕鏈CDR1，其包含SEQ ID NO: 17之胺基酸序列； (e)    輕鏈CDR2，其包含SEQ ID NO: 18之胺基酸序列；以及 (f)    輕鏈CDR3，其包含SEQ ID NO: 19之胺基酸序列。 The method of claim 72, wherein the second antibody comprises: (a) a heavy chain CDR1 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 7 to 9; (b) a heavy chain CDR2 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 10 to 13; (c) a heavy chain CDR3 comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 14 to 16; (d) a light chain CDR1 comprising the amino acid sequence of SEQ ID NO: 17; (e) a light chain CDR2 comprising the amino acid sequence of SEQ ID NO: 18; and (f) a light chain CDR3 comprising the amino acid sequence of SEQ ID NO: 19. 如請求項72之方法，其中該第二抗體包含：包含以下之序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3： (a)    分別SEQ ID NO: 7、10、14、17、18及19；或 (b)    分別SEQ ID NO: 8、11、14、17、18及19；或 (c)    分別SEQ ID NO: 9、12、15、17、18及19；或 (d)    分別SEQ ID NO: 8、13、16、17、18及19；或 (e)    分別SEQ ID NO: 8、12、16、17、18及19。 The method of claim 72, wherein the second antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the following sequences: (a) SEQ ID NOs: 7, 10, 14, 17, 18 and 19, respectively; or (b) SEQ ID NOs: 8, 11, 14, 17, 18 and 19, respectively; or (c) SEQ ID NOs: 9, 12, 15, 17, 18 and 19, respectively; or (d) SEQ ID NOs: 8, 13, 16, 17, 18 and 19, respectively; or (e) SEQ ID NOs: 8, 12, 16, 17, 18 and 19, respectively. 如請求項72之方法，其中該第二抗體包含：包含選自SEQ ID NO: 1至5之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 6之胺基酸序列的輕鏈可變區。The method of claim 72, wherein the second antibody comprises: a heavy chain variable region comprising the amino acid sequence selected from SEQ ID NO: 1 to 5; and a light comprising the amino acid sequence of SEQ ID NO: 6 chain variable region. 如請求項72之方法，其中該第二抗體包含：包含選自SEQ ID NO: 20至24之胺基酸序列的重鏈；及包含SEQ ID NO: 25之胺基酸序列的輕鏈。The method of claim 72, wherein the second antibody comprises: a heavy chain comprising the amino acid sequence selected from the group consisting of SEQ ID NO: 20 to 24; and a light chain comprising the amino acid sequence of SEQ ID NO: 25. 如請求項1至71中任一項之方法，其中該第二抗體結合CD40。The method of any one of claims 1 to 71, wherein the second antibody binds CD40. 如請求項77之方法，其中該第二抗體包含：包含以下之序列的重鏈CDR1、CDR2及CDR3以及輕鏈CDR1、CDR2及CDR3：(a)分別SEQ ID NO: 30、31、32、33、34及35；或(b)分別SEQ ID NO: 30、36、32、33、34及35。The method of claim 77, wherein the second antibody comprises: heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the sequences of: (a) SEQ ID NOs: 30, 31, 32, 33, respectively , 34 and 35; or (b) SEQ ID NOs: 30, 36, 32, 33, 34 and 35, respectively. 如請求項77之方法，其中該第二抗體包含：包含SEQ ID NO: 28之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 29之胺基酸序列的輕鏈可變區。The method of claim 77, wherein the second antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 28; and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 29 . 如請求項77之方法，其中該第二抗體包含：包含SEQ ID NO: 26之胺基酸序列的重鏈；及包含SEQ ID NO: 27之胺基酸序列的輕鏈。The method of claim 77, wherein the second antibody comprises: a heavy chain comprising the amino acid sequence of SEQ ID NO:26; and a light chain comprising the amino acid sequence of SEQ ID NO:27. 如請求項1至71中任一項之方法，其中該第二抗體結合CD70。The method of any one of claims 1 to 71, wherein the second antibody binds CD70. 如請求項81之方法，其中該第二抗體包含分別包含SEQ ID NO:53至58之序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of claim 81, wherein the second antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the sequences of SEQ ID NOs: 53 to 58, respectively. 如請求項81之方法，其中該第二抗體包含：包含SEQ ID NO: 41之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 42之胺基酸序列的輕鏈可變區。The method of claim 81, wherein the second antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 41; and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 42 . 如請求項1至71中任一項之方法，其中該第二抗體結合BCMA。The method of any one of claims 1 to 71, wherein the second antibody binds BCMA. 如請求項84之方法，其中該第二抗體包含分別包含SEQ ID NO: 47至52之序列的重鏈CDR1、CDR2及CDR3及輕鏈CDR1、CDR2及CDR3。The method of claim 84, wherein the second antibody comprises heavy chain CDR1, CDR2 and CDR3 and light chain CDR1, CDR2 and CDR3 comprising the sequences of SEQ ID NOs: 47 to 52, respectively. 如請求項84之方法，其中該第二抗體包含：包含SEQ ID NO: 45之胺基酸序列的重鏈可變區；及包含SEQ ID NO: 46之胺基酸序列的輕鏈可變區。The method of claim 84, wherein the second antibody comprises: a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 45; and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 46 . 如請求項1至86中任一項之方法，其中該第二抗體為IgG1或IgG3抗體。The method of any one of claims 1 to 86, wherein the second antibody is an IgG1 or IgG3 antibody. 如請求項1至87中任一項之方法，其中該第二抗體包含於抗體組合物中，其中該組合物中之至少90%、至少91%、至少92%、至少93%、至少94%、至少95%、至少96%、至少97%、至少98%或至少99%之該等抗體未經岩藻糖基化。The method of any one of claims 1 to 87, wherein the second antibody is included in an antibody composition, wherein at least 90%, at least 91%, at least 92%, at least 93%, at least 94% in the composition , at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of these antibodies are not fucosylated. 如請求項88之方法，其中該組合物中之每一抗體包含與該第二抗體相同的重鏈及輕鏈胺基酸序列。The method of claim 88, wherein each antibody in the composition comprises the same heavy and light chain amino acid sequences as the second antibody. 如請求項1至89中任一項之方法，其中與相同同種型之對應野生型Fc相比，該第二抗體之該Fc增強與一或多種活化FcγR之結合，其中該等活化FcγR包括FcγRIIIa、FcγRIIa及/或FcγRI中之一或多者。The method of any one of claims 1 to 89, wherein the Fc of the second antibody enhances binding to one or more activated FcγRs compared to a corresponding wild-type Fc of the same isotype, wherein the activated FcγRs comprise FcγRIIIa , one or more of FcyRIIa and/or FcyRI. 如請求項90之方法，其中該第二抗體之該Fc增強與FcγRIIIa之結合。The method of claim 90, wherein the Fc of the second antibody enhances binding to FcyRIIIa. 如請求項1至91中任一項之方法，其中與相同同種型之對應野生型Fc相比，該第二抗體之該Fc減弱與一或多種抑制性FcγR之結合。The method of any one of claims 1 to 91, wherein the Fc of the second antibody attenuates binding to one or more inhibitory FcyRs compared to a corresponding wild-type Fc of the same isotype. 如請求項92之方法，其中該第二抗體之該Fc減弱與FcγRIIb之結合。The method of claim 92, wherein the Fc of the second antibody attenuates binding to FcyRIIb. 如請求項1至93中任一項之方法，其中該第二抗體之該Fc增強與FcγRIIIa之結合且減弱與FcγRIIb之結合。The method of any one of claims 1 to 93, wherein the Fc of the second antibody enhances binding to FcyRIIIa and reduces binding to FcyRIIb. 如請求項1至94中任一項之方法，其中該第二抗體為單株抗體。The method of any one of claims 1 to 94, wherein the second antibody is a monoclonal antibody. 如請求項1至95中任一項之方法，其中該第二抗體為人類化抗體或人類抗體。The method of any one of claims 1 to 95, wherein the second antibody is a humanized antibody or a human antibody. 如請求項1至96中任一項之方法，其中該癌症為膀胱癌、乳癌、子宮癌、子宮頸癌、卵巢癌、前列腺癌、睪丸癌、食道癌、胃腸癌、胃癌(gastric cancer)、胰臟癌、大腸直腸癌、大腸癌、腎癌、透明細胞腎癌瘤、頭頸癌、肺癌、肺腺癌、胃癌(stomach cancer)、生殖細胞癌、骨癌、肝癌、甲狀腺癌、皮膚癌、黑素瘤、中樞神經系統贅瘤、間皮瘤、淋巴瘤、白血病、慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤、霍奇金氏淋巴瘤(Hodgkin lymphoma)、骨髓瘤或肉瘤。The method of any one of claims 1 to 96, wherein the cancer is bladder cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, testicular cancer, esophageal cancer, gastrointestinal cancer, gastric cancer, Pancreatic cancer, colorectal cancer, colorectal cancer, kidney cancer, clear cell renal carcinoma, head and neck cancer, lung cancer, lung adenocarcinoma, stomach cancer, germ cell cancer, bone cancer, liver cancer, thyroid cancer, skin cancer, Melanoma, CNS neoplasms, mesothelioma, lymphoma, leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, Hodgkin lymphoma, bone marrow tumor or sarcoma. 如請求項1至97中任一項之方法，其中該癌症為淋巴瘤、白血病、慢性淋巴細胞白血病、彌漫性大B細胞淋巴瘤、濾泡性淋巴瘤或霍奇金氏淋巴瘤。The method of any one of claims 1 to 97, wherein the cancer is lymphoma, leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, follicular lymphoma, or Hodgkin's lymphoma. 如請求項1至98中任一項之方法，其中該抗體-藥物結合物及該第二抗體經同時投與。The method of any one of claims 1 to 98, wherein the antibody-drug conjugate and the second antibody are administered simultaneously. 如請求項99之方法，其中該抗體-藥物結合物及該第二抗體以單一醫藥組合物形式投與。The method of claim 99, wherein the antibody-drug conjugate and the second antibody are administered as a single pharmaceutical composition. 如請求項1至98中任一項之方法，其中該抗體-藥物結合物及該第二抗體經依序投與。The method of any one of claims 1 to 98, wherein the antibody-drug conjugate and the second antibody are administered sequentially. 如請求項101之方法，其中至少該抗體-藥物結合物之第一劑量係在該第二抗體之第一劑量之前投與；或其中至少該第二抗體之第一劑量係在該抗體-藥物結合物之第一劑量之前投與。The method of claim 101, wherein at least a first dose of the antibody-drug conjugate is administered before a first dose of the second antibody; or wherein at least a first dose of the second antibody is administered before the antibody-drug The first dose of the conjugate is administered before. 如請求項1至102中任一項之方法，其中該第二抗體耗盡T調節性細胞(Treg)。The method of any one of claims 1 to 102, wherein the second antibody depletes T regulatory cells (Treg). 如請求項1至103中任一項之方法，其中該抗體-藥物結合物誘導針對表現由該抗體-藥物結合物結合之抗原的細胞的免疫記憶。The method of any one of claims 1 to 103, wherein the antibody-drug conjugate induces immune memory against cells expressing the antigen bound by the antibody-drug conjugate. 如請求項104之方法，其中該免疫記憶誘導包含誘導記憶T細胞。The method of claim 104, wherein the induction of immune memory comprises induction of memory T cells. 如請求項1至105中任一項之方法，其中該第二抗體活化抗原呈遞細胞(APC)。The method of any one of claims 1 to 105, wherein the second antibody activates antigen presenting cells (APCs). 如請求項1至106中任一項之方法，其中該第二抗體增強CD8 T細胞反應。The method of any one of claims 1 to 106, wherein the second antibody enhances CD8 T cell responses. 如請求項1至107中任一項之方法，其中該第二抗體上調共刺激受體。The method of any one of claims 1 to 107, wherein the second antibody upregulates a costimulatory receptor. 如請求項1至108中任一項之方法，其中投與該ADC及該第二抗體有助於釋放免疫活化細胞介素。The method of any one of claims 1 to 108, wherein administering the ADC and the second antibody facilitates the release of an immune-activating interleukin. 如請求項109之方法，其中該免疫活化細胞介素為CXCL10或IFNγ。The method of claim 109, wherein the immune activating interleukin is CXCL10 or IFNγ. 如請求項1至110中任一項之方法，其中該ADC及該第二抗體協同地起作用。The method of any one of claims 1 to 110, wherein the ADC and the second antibody act synergistically. 如請求項1至111中任一項之方法，其中組合投與該ADC及該第二抗體具有與以單藥療法形式投與該ADC或該第二抗體時相當的毒性概況。The method of any one of claims 1 to 111, wherein the ADC and the second antibody are administered in combination with a toxicity profile comparable to when the ADC or the second antibody is administered as monotherapy. 如請求項1至112中任一項之方法，其中該ADC及/或該第二抗體在組合給藥時之有效劑量小於在以單藥療法形式投與時之有效劑量。The method of any one of claims 1 to 112, wherein the effective dose of the ADC and/or the second antibody when administered in combination is less than the effective dose when administered as a monotherapy. 如請求項1至113中任一項之方法，其中該癌症具有高腫瘤突變負荷。The method of any one of claims 1 to 113, wherein the cancer has a high tumor mutational burden. 如請求項1至114中任一項之方法，其中該癌症具有微衛星不穩定性。The method of any one of claims 1 to 114, wherein the cancer has microsatellite instability.
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Family Cites Families (95)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4307016A (en) 1978-03-24 1981-12-22 Takeda Chemical Industries, Ltd. Demethyl maytansinoids
US4256746A (en) 1978-11-14 1981-03-17 Takeda Chemical Industries Dechloromaytansinoids, their pharmaceutical compositions and method of use
JPS55102583A (en) 1979-01-31 1980-08-05 Takeda Chem Ind Ltd 20-acyloxy-20-demethylmaytansinoid compound
JPS55162791A (en) 1979-06-05 1980-12-18 Takeda Chem Ind Ltd Antibiotic c-15003pnd and its preparation
JPS5645483A (en) 1979-09-19 1981-04-25 Takeda Chem Ind Ltd C-15003phm and its preparation
JPS5645485A (en) 1979-09-21 1981-04-25 Takeda Chem Ind Ltd Production of c-15003pnd
EP0028683A1 (en) 1979-09-21 1981-05-20 Takeda Chemical Industries, Ltd. Antibiotic C-15003 PHO and production thereof
WO1982001188A1 (en) 1980-10-08 1982-04-15 Takeda Chemical Industries Ltd 4,5-deoxymaytansinoide compounds and process for preparing same
US4450254A (en) 1980-11-03 1984-05-22 Standard Oil Company Impact improvement of high nitrile resins
US4315929A (en) 1981-01-27 1982-02-16 The United States Of America As Represented By The Secretary Of Agriculture Method of controlling the European corn borer with trewiasine
US4313946A (en) 1981-01-27 1982-02-02 The United States Of America As Represented By The Secretary Of Agriculture Chemotherapeutically active maytansinoids from Trewia nudiflora
JPS57192389A (en) 1981-05-20 1982-11-26 Takeda Chem Ind Ltd Novel maytansinoid
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
US4676980A (en) 1985-09-23 1987-06-30 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Target specific cross-linked heteroantibodies
US4946778A (en) 1987-09-21 1990-08-07 Genex Corporation Single polypeptide chain binding molecules
JP3101690B2 (en) 1987-03-18 2000-10-23 エス・ビィ・２・インコーポレイテッド Modifications of or for denatured antibodies
US5047335A (en) 1988-12-21 1991-09-10 The Regents Of The University Of Calif. Process for controlling intracellular glycosylation of proteins
CA2062795A1 (en) 1989-06-29 1990-12-30 Michael W. Fanger Bispecific reagents for aids therapy
EP0547065B1 (en) 1990-06-29 2001-08-29 Large Scale Biology Corporation Melanin production by transformed microorganisms
US5571894A (en) 1991-02-05 1996-11-05 Ciba-Geigy Corporation Recombinant antibodies specific for a growth factor receptor
US5278299A (en) 1991-03-18 1994-01-11 Scripps Clinic And Research Foundation Method and composition for synthesizing sialylated glycosyl compounds
US5587458A (en) 1991-10-07 1996-12-24 Aronex Pharmaceuticals, Inc. Anti-erbB-2 antibodies, combinations thereof, and therapeutic and diagnostic uses thereof
WO1993008829A1 (en) 1991-11-04 1993-05-13 The Regents Of The University Of California Compositions that mediate killing of hiv-infected cells
WO1993010260A1 (en) 1991-11-21 1993-05-27 The Board Of Trustees Of The Leland Stanford Junior University Controlling degradation of glycoprotein oligosaccharides by extracellular glycosisases
EP0625200B1 (en) 1992-02-06 2005-05-11 Chiron Corporation Biosynthetic binding protein for cancer marker
US5641870A (en) 1995-04-20 1997-06-24 Genentech, Inc. Low pH hydrophobic interaction chromatography for antibody purification
US5811097A (en) 1995-07-25 1998-09-22 The Regents Of The University Of California Blockade of T lymphocyte down-regulation associated with CTLA-4 signaling
US5855887A (en) 1995-07-25 1999-01-05 The Regents Of The University Of California Blockade of lymphocyte down-regulation associated with CTLA-4 signaling
US6051227A (en) 1995-07-25 2000-04-18 The Regents Of The University Of California, Office Of Technology Transfer Blockade of T lymphocyte down-regulation associated with CTLA-4 signaling
US6245347B1 (en) 1995-07-28 2001-06-12 Zars, Inc. Methods and apparatus for improved administration of pharmaceutically active compounds
DE19541260A1 (en) 1995-11-06 1997-05-07 Lohmann Therapie Syst Lts Therapeutic preparation for transdermal application of active ingredients through the skin
US6207157B1 (en) 1996-04-23 2001-03-27 The United States Of America As Represented By The Department Of Health And Human Services Conjugate vaccine for nontypeable Haemophilus influenzae
US5834597A (en) 1996-05-20 1998-11-10 Protein Design Labs, Inc. Mutated nonactivating IgG2 domains and anti CD3 antibodies incorporating the same
US6512010B1 (en) 1996-07-15 2003-01-28 Alza Corporation Formulations for the administration of fluoxetine
US6248864B1 (en) 1997-12-31 2001-06-19 Adherex Technologies, Inc. Compounds and methods and modulating tissue permeability
DK2180007T4 (en) 1998-04-20 2017-11-27 Roche Glycart Ag Glycosylation technique for antibodies to enhance antibody-dependent cell cytotoxicity
US6312717B1 (en) 1998-07-07 2001-11-06 Bristol-Myers Squibb Company Method for treatment of anxiety and depression
SE9802864D0 (en) 1998-08-27 1998-08-27 Pharmacia & Upjohn Ab Transdermally administered tolterodine as an antimuscarinic agent for the treatment of overactive bladder
EE05627B1 (en) 1998-12-23 2013-02-15 Pfizer Inc. Human monoclonal antibodies to CTLA-4
US6946129B1 (en) 1999-06-08 2005-09-20 Seattle Genetics, Inc. Recombinant anti-CD40 antibody and uses thereof
US7605238B2 (en) 1999-08-24 2009-10-20 Medarex, Inc. Human CTLA-4 antibodies and their uses
EP1792991A1 (en) 1999-08-24 2007-06-06 Medarex, Inc. Human CTLA-4 antibodies and their uses
US6544548B1 (en) 1999-09-13 2003-04-08 Keraplast Technologies, Ltd. Keratin-based powders and hydrogel for pharmaceutical applications
US6589549B2 (en) 2000-04-27 2003-07-08 Macromed, Incorporated Bioactive agent delivering system comprised of microparticles within a biodegradable to improve release profiles
CA2466279A1 (en) 2001-11-13 2003-05-22 Dana-Farber Cancer Institute, Inc. Agents that modulate immune cell activation and methods of use thereof
ES2556641T3 (en) 2002-07-31 2016-01-19 Seattle Genetics, Inc. Drug conjugates and their use to treat cancer, an autoimmune disease or an infectious disease
CA2508660C (en) 2002-12-23 2013-08-20 Wyeth Antibodies against pd-1 and uses therefor
ES2457538T3 (en) 2003-02-20 2014-04-28 Seattle Genetics, Inc. Anti-CD70-drug antibody conjugates and their use for the treatment of cannula and immune disorders
SG195524A1 (en) 2003-11-06 2013-12-30 Seattle Genetics Inc Monomethylvaline compounds capable of conjugation to ligands
BRPI0513959A (en) 2004-07-30 2008-05-20 Rinat Neuroscience Corp antibodies directed against beta-amyloid peptide, their pharmaceutical compositions, kit and methods of manufacture thereof
CA2605507C (en) 2005-04-19 2016-06-28 Seattle Genetics, Inc. Humanized anti-cd70 binding agents and uses thereof
SI2439273T1 (en) 2005-05-09 2019-05-31 Ono Pharmaceutical Co., Ltd. Human monoclonal antibodies to programmed death 1(PD-1) and methods for treating cancer using anti-PD-1 antibodies alone or in combination with other immunotherapeutics
EP1885399B1 (en) 2005-05-26 2010-10-20 Seattle Genetics, Inc. Humanized anti-cd40 antibodies and their methods of use
CN101248089A (en) 2005-07-01 2008-08-20 米德列斯公司 Human monoclonal antibodies to programmed death ligand 1(PD-L1)
US8871720B2 (en) 2005-07-07 2014-10-28 Seattle Genetics, Inc. Monomethylvaline compounds having phenylalanine carboxy modifications at the C-terminus
ES2708763T3 (en) 2005-07-07 2019-04-11 Seattle Genetics Inc Compounds of monomethylvaline that have modifications of the side chain of phenylalanine at the C-terminus
EP1790664A1 (en) 2005-11-24 2007-05-30 Ganymed Pharmaceuticals AG Monoclonal antibodies against claudin-18 for treatment of cancer
US8455622B2 (en) 2006-12-01 2013-06-04 Seattle Genetics, Inc. Variant target binding agents and uses thereof
WO2008091954A2 (en) 2007-01-23 2008-07-31 Xencor, Inc. Optimized cd40 antibodies and methods of using the same
KR101586617B1 (en) 2007-06-18 2016-01-20 머크 샤프 앤 도메 비.브이. Antibodies to human programmed death receptor PD-1
US8877688B2 (en) 2007-09-14 2014-11-04 Adimab, Llc Rationally designed, synthetic antibody libraries and uses therefor
EP3753947A1 (en) 2007-09-14 2020-12-23 Adimab, LLC Rationally designed, synthetic antibody libraries and uses therefor
US7968687B2 (en) 2007-10-19 2011-06-28 Seattle Genetics, Inc. CD19 binding agents and uses thereof
EP2262837A4 (en) 2008-03-12 2011-04-06 Merck Sharp & Dohme Pd-1 binding proteins
KR101855381B1 (en) 2008-04-09 2018-05-09 제넨테크, 인크. Novel compositions and methods for the treatment of immune related diseases
CN102076865B (en) 2008-05-02 2016-03-16 西雅图基因公司 The antibody reduced for the manufacture of core fucosylation and the method and composition of antibody derivatives
SI2342226T1 (en) 2008-09-26 2016-11-30 Dana-Farber Cancer Institute Inc. Human anti-pd-1, pd-l1, and pd-l2 antibodies and uses thereof
UA109633C2 (en) 2008-12-09 2015-09-25 HUMAN ANTIBODY AGAINST TISSUE FACTOR
UA109108C2 (en) 2008-12-09 2015-07-27 Дженентек, Інк. Anti-pd-ll antibody and its use to enhance t-cell function
ES2629337T3 (en) 2009-02-09 2017-08-08 Inserm - Institut National De La Santé Et De La Recherche Médicale Antibodies against PD-1 and antibodies against PD-L1 and uses thereof
US20130017199A1 (en) 2009-11-24 2013-01-17 AMPLIMMUNE ,Inc. a corporation Simultaneous inhibition of pd-l1/pd-l2
US20130022629A1 (en) 2010-01-04 2013-01-24 Sharpe Arlene H Modulators of Immunoinhibitory Receptor PD-1, and Methods of Use Thereof
AU2011267106B2 (en) 2010-06-15 2015-05-14 Genmab A/S Human antibody drug conjugates against tissue factor
JP2013532153A (en) 2010-06-18 2013-08-15 ザ ブリガム アンド ウィメンズ ホスピタル インコーポレイテッド Bispecific antibodies against TIM-3 and PD-1 for immunotherapy against chronic immune disease
US8907053B2 (en) 2010-06-25 2014-12-09 Aurigene Discovery Technologies Limited Immunosuppression modulating compounds
MX360336B (en) 2010-07-16 2018-10-30 Adimab Llc Star Antibody libraries.
AU2011312417B2 (en) 2010-09-29 2015-08-20 Agensys, Inc. Antibody drug conjugates (ADC) that bind to 191P4D12 proteins
UA112434C2 (en) 2011-05-27 2016-09-12 Ґлаксо Ґруп Лімітед ANTIGENCY BINDING SPECIFICALLY Binds to ALL
KR20240034882A (en) 2013-10-15 2024-03-14 씨젠 인크. Pegylated drug-linkers for improved ligand-drug conjugate pharmacokinetics
MA40475A (en) 2014-08-19 2017-06-28 Fayadat Dilman Laurence Anti-tigit antibodies
ES2886884T3 (en) 2014-09-11 2021-12-21 Seagen Inc Targeted Delivery of Pharmaceutical Substances Containing Tertiary Amines
EP3212230B1 (en) 2014-10-29 2021-01-20 Seagen Inc. Dosage and administration of non-fucosylated anti-cd40 antibodies
AU2015369683B2 (en) 2014-12-23 2020-12-10 Bristol-Myers Squibb Company Antibodies to TIGIT
TWI715587B (en) 2015-05-28 2021-01-11 美商安可美德藥物股份有限公司 Tigit-binding agents and uses thereof
AU2016325610B2 (en) 2015-09-25 2019-10-10 Genentech, Inc. Anti-TIGIT antibodies and methods of use
SG11201803213XA (en) 2015-11-12 2018-05-30 Siamab Therapeutics Inc Glycan-interacting compounds and methods of use
EP4074328A1 (en) 2015-12-04 2022-10-19 Seagen Inc. Intermediates of conjugates of quaternized tubulysin compounds
MX2018009700A (en) 2016-02-17 2019-01-24 Seattle Genetics Inc Bcma antibodies and use of same to treat cancer and immunological disorders.
CN113817066A (en) * 2018-04-13 2021-12-21 艾菲默德有限责任公司 Natural killer cell-conjugated antibody fusion constructs
JP7273858B2 (en) * 2018-06-15 2023-05-15 シャンハイ ミラコーケン インコーポレイティド Methods and materials for treating cancer
AU2019415848A1 (en) 2018-12-28 2021-08-19 Nanjing GenScript Biotech Co., Ltd. Claudin18.2 binding moieties and uses thereof
EP3920968A1 (en) 2019-02-05 2021-12-15 Seagen Inc. Anti-cd228 antibodies and antibody-drug conjugates
CN114630840A (en) 2019-08-20 2022-06-14 苏州创胜医药集团有限公司 Novel anti-CLDN 18.2 antibodies
WO2021055865A1 (en) 2019-09-19 2021-03-25 Seagen Inc. Selective drug release from internalized conjugates of biologically active compounds
CN110818795B (en) * 2020-01-10 2020-04-24 上海复宏汉霖生物技术股份有限公司 anti-TIGIT antibodies and methods of use

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