The use of CrossMAb technology for the generation of bi- and multispecific antibodies

mAbs

Volumn 8, Issue 6, 2016, Pages 1010-1020

  Klein, Christian ^a   Schaefer, Wolfgang ^b   Regula, Jörg T ^b  

^a Roche Pharmaceutical Research Early Development   (Switzerland)

^b ROCHE INNOVATION CENTER MUNICH   (Germany)

Author keywords

2+1, 1+1; 2+2; Ang 2; asymmetric; CEA TCB; CrossMAb; DAF CrossMAb; DuoMAb; DVD CrossMAb; EGFR; HER1; HER3; heterodimeric; Immunoglobulin domain crossover; Kappa Lambda CrossMAb; knobs into holes (KiH); MoAb; MoAb Dimer; MonoMAb; P329G LALA; RG7221; RG7386; RG7716; RG7802; Triple A; vanucizumab; VEGF A

Indexed keywords

ANGIOPOIETIN 2; ATEZOLIZUMAB; BEVACIZUMAB; BISPECIFIC ANTIBODY; IMMUNOGLOBULIN G ANTIBODY; PERTUZUMAB; RANIBIZUMAB; TRASTUZUMAB; VANUCIZUMAB; VASCULOTROPIN A; ANTINEOPLASTIC AGENT; IMMUNOGLOBULIN F(AB) FRAGMENT; IMMUNOGLOBULIN G; MONOCLONAL ANTIBODY;

ANTIBODY DEPENDENT CELLULAR CYTOTOXICITY; CLINICAL TRIAL (TOPIC); COMPLEMENT DEPENDENT CYTOTOXICITY; DRUG EFFICACY; DRUG RESEARCH; DRUG SCREENING; HUMAN; LASER-INDUCED CHOROIDAL NEOVASCULARIZATION; OVARY CANCER; PHAGOCYTOSIS; PHARMACODYNAMIC PARAMETERS; PHARMACOKINETIC PARAMETERS; PHASE 1 CLINICAL TRIAL (TOPIC); PROTEIN STRUCTURE; REVIEW; STATIC ELECTRICITY; TREATMENT OUTCOME; TUMOR VOLUME; TUMOR XENOGRAFT; CHEMISTRY; IMMUNOLOGY; PROCEDURES; PROTEIN ENGINEERING; PROTEIN MULTIMERIZATION;

ANTIBODIES, BISPECIFIC; ANTIBODIES, MONOCLONAL; ANTINEOPLASTIC AGENTS; HUMANS; IMMUNOGLOBULIN FAB FRAGMENTS; IMMUNOGLOBULIN G; PROTEIN ENGINEERING; PROTEIN MULTIMERIZATION; STATIC ELECTRICITY;

EID: 84979224082     PISSN: 19420862     EISSN: 19420870     Source Type: Journal    
DOI: 10.1080/19420862.2016.1197457     Document Type: Review

References (76)

1. W.Schaefer, H.R.Volger, S.Lorenz, S.Imhof-Jung, J.T.Regula, C.Klein, M.Molhoj. Heavy and light chain pairing of bivalent quadroma and knobs-into-holes antibodies analyzed by uhr-esi-qtof mass spectrometry. MAbs 2016; 8(1):49-55; PMID:26496506; doi 10.1080/19420862.2015.1111498
2. C.Klein, C.Sustmann, M.Thomas, K.Stubenrauch, R.Croasdale, J.Schanzer, U.Brinkmann, H.Kettenberger, J.T.Regula, W.Schaefer. Progress in overcoming the chain association issue in bispecific heterodimeric igg antibodies. MAbs 2012; 4(6):653-63; PMID:22925968; doi 10.4161/mabs.21379
3. A.M.Merchant, Z.Zhu, J.Q.Yuan, A.Goddard, C.W.Adams, L.G.Presta, P.Carter. An efficient route to human bispecific igg. Nat Biotechnol 1998; 16(7):677-81; PMID:9661204; doi 10.1038/nbt0798-677
4. W.Schaefer, J.T.Regula, M.Bahner, J.Schanzer, R.Croasdale, H.Durr, C.Gassner, G.Georges, H.Kettenberger, S.Imhof-Jung, et al. Immunoglobulin domain crossover as a generic approach for the production of bispecific igg antibodies. Proc Natl Acad Sci U S A 2011; 108(27):11187-92; PMID:21690412; doi 10.1073/pnas.1019002108
5. S.Dengl, M.Wehmer, F.Hesse, F.Lipsmeier, O.Popp, K.Lang. Aggregation and chemical modification of monoclonal antibodies under upstream processing conditions. Pharm Res 2013; 30(5):1380-99; PMID:23322133; doi 10.1007/s11095-013-0977-8
6. J.Gu, J.Yang, Q.Chang, Z.Liu, T.Ghayur. Identification of anti-egfr and anti-erbb3 dual variable domains immunoglobulin (dvd-ig) proteins with unique activities. PLoS One 2015; 10(5):e0124135; PMID:25997020; doi 10.1371/journal.pone.0124135
7. I.Correia, J.Sung, R.Burton, C.G.Jakob, B.Carragher, T.Ghayur, C.Radziejewski. The structure of dual-variable-domain immunoglobulin molecules alone and bound to antigen. MAbs 2013; 5(3):364-72; PMID:23572180; doi 10.4161/mabs.24258
8. C.G.Jakob, R.Edalji, R.A.Judge, E.DiGiammarino, Y.Li, J.Gu, T.Ghayur. Structure reveals function of the dual variable domain immunoglobulin (dvd-ig) molecule. MAbs 2013; 5(3):358-63; PMID:23549062; doi 10.4161/mabs.23977
9. C.Wu, H.Ying, C.Grinnell, S.Bryant, R.Miller, A.Clabbers, S.Bose, D.McCarthy, R.R.Zhu, L.Santora, R.Davis-Taber, et al. Simultaneous targeting of multiple disease mediators by a dual-variable-domain immunoglobulin. Nat Biotechnol 2007; 25(11):1290-7; PMID:17934452; doi 10.1038/nbt1345
10. A.Steinmetz, F.Vallee, C.Beil, C.Lange, N.Baurin, J.Beninga, C.Capdevila, C.Corvey, A.Dupuy, P.Ferrari, et al. Codv-ig, a universal bispecific tetravalent and multifunctional immunoglobulin format for medical applications. MAbs 2016 July; 8(5): 867—78; PMID:26984268; doi 10.1080/19420862.2016.1162932
11. E.J.Smith, K.Olson, L.J.Haber, B.Varghese, P.Duramad, A.D.Tustian, A.Oyejide, J.R.Kirshner, L.Canova, J.Menon, et al. A novel, native-format bispecific antibody triggering t-cell killing of b-cells is robustly active in mouse tumor models and cynomolgus monkeys. Sci Rep 2015; 5:17943; PMID:26659273; doi 10.1038/srep17943
12. N.Fischer, G.Elson, G.Magistrelli, E.Dheilly, N.Fouque, A.Laurendon, F.Gueneau, U.Ravn, J.F.Depoisier, V.Moine, et al. Exploiting light chains for the scalable generation and platform purification of native human bispecific igg. Nat Commun 2015; 6:6113; PMID:25672245; doi 10.1038/ncomms7113
13. Z.Sampei, T.Igawa, T.Soeda, Y.Okuyama-Nishida, C.Moriyama, T.Wakabayashi, E.Tanaka, A.Muto, T.Kojima, T.Kitazawa, et al. Identification and multidimensional optimization of an asymmetric bispecific igg antibody mimicking the function of factor viii cofactor activity. PLoS One 2013; 8(2):e57479; PMID:23468998; doi 10.1371/journal.pone.0057479
14. T.Kitazawa, T.Igawa, Z.Sampei, A.Muto, T.Kojima, T.Soeda, K.Yoshihashi, Y.Okuyama-Nishida, H.Saito, H.Tsunoda, et al. A bispecific antibody to factors ixa and x restores factor viii hemostatic activity in a hemophilia a model. Nat Med 2012; 18(10):1570-4; PMID:23023498; doi 10.1038/nm.2942
15. A.D.Tustian, C.Endicott, B.Adams, J.Mattila, H.Bak. Development of purification processes for fully human bispecific antibodies based upon modification of protein a binding avidity. MAbs 2016; 8(4):828-38; PMID:26963837; doi 10.1080/19420862.2016.1160192
16. C.Spiess, M.Merchant, A.Huang, Z.Zheng, N.Y.Yang, J.Peng, D.Ellerman, W.Shatz, D.Reilly, D.G.Yansura, et al. Bispecific antibodies with natural architecture produced by co-culture of bacteria expressing two distinct half-antibodies. Nat Biotechnol 2013; 31(8):753-8; PMID:23831709; doi 10.1038/nbt.2621
17. A.F.Labrijn, J.I.Meesters, B.E.de Goeij, E.T.van den Bremer, J.Neijssen, M.D.van Kampen, K.Strumane, S.Verploegen, A.Kundu, M.J.Gramer, et al. Efficient generation of stable bispecific igg1 by controlled fab-arm exchange. Proc Natl Acad Sci U S A 2013; 110(13):5145-50; PMID:23479652; doi 10.1073/pnas.1220145110
18. C.Spiess, J.Bevers, 3rd, J.Jackman, N.Chiang, G.Nakamura, M.Dillon, H.Liu, P.Molina, J.M.Elliott, W.Shatz, et al. Development of a human igg4 bispecific antibody for dual targeting of interleukin-4 (il-4) and interleukin-13 (il-13) cytokines. J Biol Chem 2013; 288(37):26583-93; PMID:23880771; doi 10.1074/jbc.M113.480483
19. P.Strop, W.H.Ho, L.M.Boustany, Y.N.Abdiche, K.C.Lindquist, S.E.Farias, M.Rickert, C.T.Appah, E.Pascua, T.Radcliffe, et al. Generating bispecific human igg1 and igg2 antibodies from any antibody pair. J Mol Biol 2012; 420(3):204-19; PMID:22543237; doi 10.1016/j.jmb.2012.04.020
20. G.Schaefer, L.Haber, L.M.Crocker, S.Shia, L.Shao, D.Dowbenko, K.Totpal, A.Wong, C.V.Lee, S.Stawicki, et al. A two-in-one antibody against her3 and egfr has superior inhibitory activity compared with monospecific antibodies. Cancer Cell 2011; 20(4):472-86; PMID:22014573; doi 10.1016/j.ccr.2011.09.003
21. R.Beckmann. Wo2012163520: Dual targeting. 2012
22. Y.Mazor, V.Oganesyan, C.Yang, A.Hansen, J.Wang, H.Liu, K.Sachsenmeier, M.Carlson, D.V.Gadre, M.J.Borrok, et al. Improving target cell specificity using a novel monovalent bispecific igg design. MAbs 2015; 7(2):377-89; PMID:25621507; doi 10.1080/19420862.2015.1007816
23. Z.Liu, E.C.Leng, K.Gunasekaran, M.Pentony, M.Shen, M.Howard, J.Stoops, K.Manchulenko, V.Razinkov, H.Liu, et al. A novel antibody engineering strategy for making monovalent bispecific heterodimeric igg antibodies by electrostatic steering mechanism. J Biol Chem 2015; 290(12):7535-62; PMID:25583986; doi 10.1074/jbc.M114.620260
24. X.Wu, A.J.Sereno, F.Huang, S.M.Lewis, R.L.Lieu, C.Weldon, C.Torres, C.Fine, M.A.Batt, J.R.Fitchett, et al. Fab-based bispecific antibody formats with robust biophysical properties and biological activity. MAbs 2015; 7(3):470-82; PMID:25774965; doi 10.1080/19420862.2015.1022694
25. S.M.Lewis, X.Wu, A.Pustilnik, A.Sereno, F.Huang, H.L.Rick, G.Guntas, A.Leaver-Fay, E.M.Smith, C.Ho, et al. Generation of bispecific igg antibodies by structure-based design of an orthogonal fab interface. Nat Biotechnol 2014; 32(2):191-8; PMID:24463572; doi 10.1038/nbt.2797
26. R.E.Kontermann, U.Brinkmann. Bispecific antibodies. Drug Discov Today 2015; 20(7):838-47; PMID:25728220; doi 10.1016/j.drudis.2015.02.008
27. C.Spiess, Q.Zhai, P.J.Carter. Alternative molecular formats and therapeutic applications for bispecific antibodies. Mol Immunol 2015; 67(2 Pt A):95-106; PMID:25637431; doi 10.1016/j.molimm.2015.01.003
28. M.Grote, A.K.Haas, C.Klein, W.Schaefer, U.Brinkmann. Bispecific antibody derivatives based on full-length igg formats. Methods Mol Biol 2012; 901 247-63; PMID:22723106; doi 10.1007/978-1-61779-931-0_16
29. K.Gunasekaran, M.Pentony, M.Shen, L.Garrett, C.Forte, A.Woodward, S.B.Ng, T.Born, M.Retter, K.Manchulenko, et al. Enhancing antibody fc heterodimer formation through electrostatic steering effects: Applications to bispecific molecules and monovalent igg. J Biol Chem 2010; 285(25):19637-46; PMID:20400508; doi 10.1074/jbc.M110.117382
30. G.L.Moore, C.Bautista, E.Pong, D.H.Nguyen, J.Jacinto, A.Eivazi, U.S.Muchhal, S.Karki, S.Y.Chu, G.A.Lazar. A novel bispecific antibody format enables simultaneous bivalent and monovalent co-engagement of distinct target antigens. MAbs 2011; 3(6):546-57; PMID:22123055; doi 10.4161/mabs.3.6.18123
31. J.H.Davis, C.Aperlo, Y.Li, E.Kurosawa, Y.Lan, K.M.Lo, J.S.Huston. Seedbodies: Fusion proteins based on strand-exchange engineered domain (seed) ch3 heterodimers in an fc analogue platform for asymmetric binders or immunofusions and bispecific antibodies. Protein Eng Des Sel 2010; 23(4):195-202; PMID:20299542; doi 10.1093/protein/gzp094
32. M.Thomas, Y.Kienast, W.Scheuer, M.Bahner, K.Kaluza, C.Gassner, F.Herting, U.Brinkmann, S.Seeber, A.Kavlie, et al. A novel angiopoietin-2 selective fully human antibody with potent anti-tumoral and anti-angiogenic efficacy and superior side effect profile compared to pan-angiopoietin-1/-2 inhibitors. PLoS One 2013; 8(2):e54923; PMID:23405099; doi 10.1371/journal.pone.0054923
33. Y.Kienast, C.Klein, W.Scheuer, R.Raemsch, E.Lorenzon, D.Bernicke, F.Herting, S.Yu, H.H.The, L.Martarello, et al. Ang-2-vegf-a crossmab, a novel bispecific human igg1 antibody blocking vegf-a and ang-2 functions simultaneously, mediates potent antitumor, antiangiogenic, and antimetastatic efficacy. Clin Cancer Res 2013; 19(24):6730-40; PMID:24097868; doi 10.1158/1078-0432.CCR-13-0081
34. C.Gassner, F.Lipsmeier, P.Metzger, H.Beck, A.Schnueriger, J.T.Regula, J.Moelleken. Development and validation of a novel spr-based assay principle for bispecific molecules. J Pharm Biomed Anal 2015; 102:144-9; PMID:25277666; doi 10.1016/j.jpba.2014.09.007
35. K.Srivastava, J.Hu, C.Korn, S.Savant, M.Teichert, S.S.Kapel, M.Jugold, E.Besemfelder, M.Thomas, M.Pasparakis, et al. Postsurgical adjuvant tumor therapy by combining anti-angiopoietin-2 and metronomic chemotherapy limits metastatic growth. Cancer Cell 2014; 26(6):880-95; PMID:25490450; doi 10.1016/j.ccell.2014.11.005
36. H.Bessho, B.Wong, D.Huang, J.Tan, C.K.Ong, M.Iwamura, S.Hart, M.Dangl, M.Thomas, B.T.Teh. Effect of ang-2-vegf-a bispecific antibody in renal cell carcinoma. Cancer Invest 2015; 33(8):378-86; PMID:26115098; doi 10.3109/07357907.2015.1047505
37. S.Fenn, C.B.Schiller, J.J.Griese, H.Duerr, S.Imhof-Jung, C.Gassner, J.Moelleken, J.T.Regula, W.Schaefer, M.Thomas, et al. Crystal structure of an anti-ang2 crossfab demonstrates complete structural and functional integrity of the variable domain. PLoS One 2013; 8(4):e61953; PMID:23613981; doi 10.1371/journal.pone.0061953
38. J.Kloepper, L.Riedemann, Z.Amoozgar, G.Seano, K.Susek, V.Yu, N.Dalvie, R.L.Amelung, M.Datta, J.W.Song, et al. Ang-2/vegf bispecific antibody reprograms macrophages and resident microglia to anti-tumor phenotype and prolongs glioblastoma survival. Proc Natl Acad Sci U S A 2016; 113:4476-81; PMID:27044098
39. M.Hidalgo, C.Le Tourneau, C.Massard, V.Boni, E.Calvo, J.Albanell, A.Taus, M.P.Sablin, A.Varga, R.Bahleda, et al. Results from the first-in-human (fih) phase i study of ro5520985 (rg7221), a novel bispecific human anti-ang-2/anti-vegf-a antibody, administered as an intravenous infusion to patients with advanced solid tumors. J Clin Oncol 2014; 32(15)
40. A.Oaknin, A.Floquet, C.Le Tourneau, I.L.Ray-Coquard, F.Joly, M.Hidalgo, A.Leary, O.Krieter, A.Lahr, S.Rossomanno, et al. Single agent vanucizumab (ro5520985) for platinum (pt)-resistant recurrent ovarian cancer (oc): Results from a single arm extension phase of the phase i fih study. J Clin Oncol 2015; 33(15)
41. A.Otani, H.Takagi, H.Oh, S.Koyama, M.Matsumura, Y.Honda. Expressions of angiopoietins and tie2 in human choroidal neovascular membranes. Invest Ophthalmol Vis Sci 1999; 40(9):1912-20; PMID:10440243
42. H.Takagi, S.Koyama, H.Seike, H.Oh, A.Otani, M.Matsumura, Y.Honda. Potential role of the angiopoietin/tie2 system in ischemia-induced retinal neovascularization. Invest Ophthalmol Vis Sci 2003; 44(1):393-402; PMID:12506101; doi 10.1167/iovs.02-0276
43. E.S.Rennel, J.T.Regula, S.J.Harper, M.Thomas, C.Klein, D.O.Bates. A human neutralizing antibody specific to ang-2 inhibits ocular angiogenesis. Microcirculation 2011; 18(7):598-607; PMID:21851472; doi 10.1111/j.1549-8719.2011.00120.x
44. F.Zhang, J.Zhang, M.Liu, L.Zhao, R.LingHu, F.Feng, X.Gao, S.Jiao, Y.Hu, J.Yang. Combating her2-overexpressing breast cancer through induction of calreticulin exposure by tras-permut crossmab. Oncoimmunology 2015; 4(3):e994391; PMID:25949918; doi 10.4161/2162402X.2014.994391
45. L.Zhao, F.Xie, X.Tong, H.Li, Y.Chen, W.Qian, S.Duan, J.Zheng, Z.Zhao, B.Li, et al. Combating non-hodgkin lymphoma by targeting both cd20 and hla-dr through cd20-243 crossmab. MAbs 2014; 6(3):740-8; PMID:24670986; doi 10.4161/mabs.28613
46. L.Zhao, Q.Tong, W.Qian, B.Li, D.Zhang, T.Fu, S.Duan, X.Zhang, J.Zhao, J.Dai, et al. Eradication of non-hodgkin lymphoma through the induction of tumor-specific t-cell immunity by cd20-flex bifp. Blood 2013; 122(26):4230-6; PMID:24178967; doi 10.1182/blood-2013-04-496554
47. M.Asokan, R.S.Rudicell, M.Louder, K.McKee, S.O'Dell, G.Stewart-Jones, K.Wang, L.Xu, X.Chen, M.Choe, et al. Bispecific antibodies targeting different epitopes on the hiv-1 envelope exhibit broad and potent neutralization. J Virol 2015; 89(24):12501-12; PMID:26446600; doi 10.1128/JVI.02097-15
48. Y.Huang, J.Yu, A.Lanzi, X.Yao, C.D.Andrews, L.Tsai, M.Gajjar, M.Sun, M.Seaman, N.N.Padte, et al. Engineered bispecific antibodies with exquisite hiv-1-neutralizing activity. Cell 2016 June 16; 165(7):1621-1631; PMID:27315479; doi 10.1016/j.cell.2016.05.024
49. Bispecific Anti-HIV-1 Antibodies with Enhanced Breadth and Potency. Bournazos S, Gazumyan A, Seaman MS, Nussenzweig MC, Ravetch JV. Cell. 2016 Jun 16;165(7):1609-1620. doi:10.1016/j.cell.2016.04.050. PMID: 27315478.
50. H.Y.Tung, Y.C.Su, B.M.Chen, P.A.Burnouf, W.C.Huang, K.H.Chuang, Y.T.Yan, T.L.Cheng, S.R.Roffler. Selective delivery of pegylated compounds to tumor cells by anti-peg hybrid antibodies. Mol Cancer Ther 2015; 14(6):1317-26; PMID:25852063; doi 10.1158/1535-7163.MCT-15-0151
51. S.Lehmann, R.Perera, H.P.Grimm, J.Sam, S.Colombetti, T.Fauti, L.Fahrni, T.Schaller, A.Freimoser-Grundschober, J.Zielonka, et al. In vivo imaging of the activity of cea tcb, a novel t-cell bispecific antibody, reveals specific tumor targeting and fast induction of t-cell mediated tumor killing. Clin Cancer Res 2016 Apr 26; PMID:27117182
52. M.Bacac, T.Fauti, J.Sam, S.Colombetti, T.Weinzierl, D.Ouaret, W.Bodmer, S.Lehmann, T.Hofer, R.J.Hosse, et al. A novel carcinoembryonic antigen t-cell bispecific antibody (cea tcb) for the treatment of solid tumors. Clin Cancer Res 2016 Feb 9; PMID:26861458
53. M.D.Vu, S.Moser, C.Delon, M.Latzko, R.Gianotti, R.Luoend, C.Friang, R.Murr, L.J.Duerner, T.Weinzierl, et al. A new class of t-cell bispecific antibodies for the treatment of multiple myeloma, binding to b cell maturation antigen and cd3 and showing potent, specific antitumor activity in myeloma cells and long duration of action in cynomolgus monkeys. Blood 2015; 126(23)
54. A.Seckinger, J.A.Delgado, L.Moreno, B.Neuber, A.Grab, S.Lipp, J.Merino, M.D.Vu, K.Strein, F.Prosper, et al. Target expression, preclinical activity and mechanism of action of em801: A novel first-in-class bcma t-cell bispecific antibody for the treatment of multiple myeloma. Blood 2015; 126(23); PMID:NOT_FOUND
55. S.Imhof-Jung, C.Klein, J.Regula, W.Schaefer, J.Schanzer. Wo2010145792: Bispecific antigen binding proteins. 2010
56. S.Imhof-Jung, C.Klein, J.Regula, W.Schaefer, J.Schanzer. Wo2010145793: Bispecific, tetravalent antigen binding proteins. 2010
57. P.Brunker, K.Wartha, T.Friess, S.Grau-Richards, I.Waldhauer, C.Ferrara Koller, B.Weiser, M.Majety, V.Runza, H.Niu, et al. Rg7386, a novel tetravalent fap-dr5 antibody, effectively triggers fap-dependent, avidity-driven dr5 hyperclustering and tumor cell apoptosis. Mol Cancer Ther 2016; 15:946-57; PMID:27037412
58. J.M.Kim, A.Ashkenazi. Fcgamma receptors enable anticancer action of proapoptotic and immune-modulatory antibodies. J Exp Med 2013; 210(9):1647-51; PMID:23980122; doi 10.1084/jem.20131625
59. J.A.Fischer, A.J.Hueber, S.Wilson, M.Galm, W.Baum, C.Kitson, J.Auer, S.H.Lorenz, J.Moelleken, M.Bader, et al. Combined inhibition of tumor necrosis factor alpha and interleukin-17 as a therapeutic opportunity in rheumatoid arthritis: Development and characterization of a novel bispecific antibody. Arthritis Rheumatol 2015; 67(1):51-62; PMID:25303306; doi 10.1002/art.38896
60. B.Bossenmaier, H.Kettenberger, C.Klein, K.P.Kuenkele, J.Regula, W.Schaefer, M.Schwaiger, C.Sustmann. Wo2012116927: Monovalent antigen binding protein. 2012
61. B.Bossenmaier, H.Kettenberger, C.Klein, K.P.Kuenkele, J.Regula, W.Schaefer, M.Schwaiger, C.Sustmann. Wo2012116926a1: Antigen binding proteins. 2012
62. J.Bostrom, S.F.Yu, D.Kan, B.A.Appleton, C.V.Lee, K.Billeci, W.Man, F.Peale, S.Ross, C.Wiesmann, et al. Variants of the antibody herceptin that interact with her2 and vegf at the antigen binding site. Science 2009; 323(5921):1610-4; PMID:19299620; doi 10.1126/science.1165480
63. F.Castoldi, A.Hass, C.Klein, W.Schaefer, C.Sustmann. Wo2013174873a1: Multispecific antibodies. 2013
64. U.Brinkmann, R.Croasdale, E.Hoffmann, C.Klein, E.Moessner, J.Schanzer, P.Umana. Wo2010115589: Trivalent bispecific antibodies. 2010
65. S.Metz, C.Panke, A.K.Haas, J.Schanzer, W.Lau, R.Croasdale, E.Hoffmann, B.Schneider, J.Auer, C.Gassner, et al. Bispecific antibody derivatives with restricted binding functionalities that are activated by proteolytic processing. Protein Eng Des Sel 2012; 25(10):571-80; PMID:22976197; doi 10.1093/protein/gzs064
66. R.Castoldi, V.Ecker, L.Wiehle, M.Majety, R.Busl-Schuller, M.Asmussen, A.Nopora, U.Jucknischke, F.Osl, S.Kobold, et al. A novel bispecific egfr/met antibody blocks tumor-promoting phenotypic effects induced by resistance to egfr inhibition and has potent antitumor activity. Oncogene 2013; 32(50):5593-5601; PMID:23812422; doi 10.1038/onc.2013.245
67. R.Castoldi, U.Jucknischke, L.P.Pradel, E.Arnold, C.Klein, S.Scheiblich, G.Niederfellner, C.Sustmann. Molecular characterization of novel trispecific erbb-cmet-igf1r antibodies and their antigen-binding properties. Protein Eng Des Sel 2012; 25(10):551-9; PMID:22936109; doi 10.1093/protein/gzs048
68. J.M.Schanzer, K.Wartha, R.Croasdale, S.Moser, K.P.Kunkele, C.Ries, W.Scheuer, H.Duerr, S.Pompiati, J.Pollman, et al. A novel glycoengineered bispecific antibody format for targeted inhibition of epidermal growth factor receptor (egfr) and insulin-like growth factor receptor type i (igf-1r) demonstrating unique molecular properties. J Biol Chem 2014; 289(27):18693-706; PMID:24841203; doi 10.1074/jbc.M113.528109
69. R.Croasdale, C.Klein, W.Schaefer, J.Schanzer. Wo2010136172: Tri- or tetraspecific antibodies. 2010
70. S.Hu, W.Fu, W.Xu, Y.Yang, M.Cruz, S.D.Berezov, D.Jorissen, H.Takeda, W.Zhu. Four-in-one antibodies have superior cancer inhibitory activity against egfr, her2, her3, and vegf through disruption of her/met crosstalk. Cancer Res 2015; 75(1):159-70; PMID:25371409; doi 10.1158/0008-5472.CAN-14-1670
71. C.Mirschberger, C.B.Schiller, M.Schraml, N.Dimoudis, T.Friess, C.A.Gerdes, U.Reiff, V.Lifke, G.Hoelzlwimmer, I.Kolm, et al. Rg7116, a therapeutic antibody that binds the inactive her3 receptor and is optimized for immune effector activation. Cancer Res 2013; 73(16):5183-94; PMID:23780344; doi 10.1158/0008-5472.CAN-13-0099
72. G.Meneses-Lorente, T.Friess, I.Kolm, G.Holzlwimmer, S.Bader, C.Meille, M.Thomas, B.Bossenmaier. Preclinical pharmacokinetics, pharmacodynamics, and efficacy of rg7116: A novel humanized, glycoengineered anti-her3 antibody. Cancer Chemother Pharmacol 2015; 75(4):837-50; PMID:25702049; doi 10.1007/s00280-015-2697-8
73. P.Bruenker, C.Jaeger, C.Klein, E.Moessner, W.Schaefer. Wo2015052230a1: Multispecific domain exchanged common variable light chain antibodies. 2015
74. P.Bruenker, T.Fauti, C.Jaeger, C.Klein, P.Umana. Wo2014056783a1: Fc-free antibodies comprising two fab fragments and methods of use. 2014
75. H.S.Cho, K.Mason, K.X.Ramyar, A.M.Stanley, S.B.Gabelli, D.W.Denney, Jr., D.J.Leahy. Structure of the extracellular region of her2 alone and in complex with the herceptin fab. Nature 2003; 421(6924):756-60; PMID:12610629; doi 10.1038/nature01392
76. F.A.Saul, R.J.Poljak. Crystal structure of human immunoglobulin fragment fab new refined at 2.0 a resolution. Proteins 1992; 14(3):363-71; PMID:1438175; doi 10.1002/prot.340140305