title: Development of a two-part strategy to identify a therapeutic human bispecific antibody that inhibits IgE receptor signaling.
abstract: The development of bispecific antibodies as therapeutic agents for human diseases has great clinical potential, but broad application has been hindered by the difficulty of identifying bispecific antibody formats that exhibit favorable pharmacokinetic properties and ease of large-scale manufacturing. Previously, the development of an antibody technology utilizing heavy chain knobs-into-holes mutations and a single common light chain enabled the small-scale generation of human full-length bispecific antibodies. Here we have extended the technology by developing a two-part bispecific antibody discovery strategy that facilitates proof-of-concept studies and clinical candidate antibody generation. Our scheme consists of the efficient small-scale generation of bispecific antibodies lacking a common light chain and the hinge disulfides for proof-of-concept studies coupled with the identification of a common light chain bispecific antibody for large-scale production with high purity and yield. We have applied this technology to generate a bispecific antibody suitable for development as a human therapeutic. This antibody directly inhibits the activation of the high affinity IgE receptor FcepsilonRI on mast cells and basophils by cross-linking FcepsilonRI with the inhibitory receptor FcgammaRIIb, an approach that has strong therapeutic potential for asthma and other allergic diseases. Our approach for producing human bispecific full-length antibodies enables the clinical application of bispecific antibodies to a validated therapeutic pathway in asthma.
authors.affiliation: Department of Immunology, Genentech, Inc., South San Francisco, CA 94080, USA.
authors.forename: Janet
authors.initials: J
authors.lastname: Jackman
authors.forename: Yongmei
authors.initials: Y
authors.lastname: Chen
authors.forename: Arthur
authors.initials: A
authors.lastname: Huang
authors.forename: Barbara
authors.initials: B
authors.lastname: Moffat
authors.forename: Justin M
authors.initials: JM
authors.lastname: Scheer
authors.forename: Steven R
authors.initials: SR
authors.lastname: Leong
authors.forename: Wyne P
authors.initials: WP
authors.lastname: Lee
authors.forename: Juan
authors.initials: J
authors.lastname: Zhang
authors.forename: Navneet
authors.initials: N
authors.lastname: Sharma
authors.forename: Yanmei
authors.initials: Y
authors.lastname: Lu
authors.forename: Suhasini
authors.initials: S
authors.lastname: Iyer
authors.forename: Robert L
authors.initials: RL
authors.lastname: Shields
authors.forename: Nancy
authors.initials: N
authors.lastname: Chiang
authors.forename: Michele C
authors.initials: MC
authors.lastname: Bauer
authors.forename: Diana
authors.initials: D
authors.lastname: Wadley
authors.forename: Merone
authors.initials: M
authors.lastname: Roose-Girma
authors.forename: Richard
authors.initials: R
authors.lastname: Vandlen
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Yansura
authors.forename: Yan
authors.initials: Y
authors.lastname: Wu
authors.forename: Lawren C
authors.initials: LC
authors.lastname: Wu
mesh_terms: D019943:Amino Acid Substitution; D000818:Animals; D018033:Antibodies, Bispecific / Q000235:genetics / Q000627:therapeutic use*; D000918:Antibody Specificity; D001491:Basophils / Q000276:immunology; D045744:Cell Line, Tumor; D003062:Codon / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005982:Glutathione Transferase / Q000235:genetics; D006801:Humans; D007073:Immunoglobulin E / Q000235:genetics / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D051379:Mice; D016513:Mice, SCID; D010323:Passive Cutaneous Anaphylaxis / Q000276:immunology; D017455:Receptors, IgE / Q000037:antagonists & inhibitors / Q000187:drug effects / Q000276:immunology / Q000502:physiology*; D017452:Receptors, IgG / Q000276:immunology; D011994:Recombinant Proteins / Q000627:therapeutic use; D019572:Retinal Neoplasms / Q000276:immunology; D012175:Retinoblastoma / Q000276:immunology; D012680:Sensitivity and Specificity
chemical_list: D018033:Antibodies, Bispecific; D003062:Codon; C499173:FCGR2B protein, human; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D017455:Receptors, IgE; D017452:Receptors, IgG; D011994:Recombinant Proteins; D007073:Immunoglobulin E; D005982:Glutathione Transferase
references.citation: J Immunol Methods. 2001 Feb 1;248(1-2):7-15
references.pmid: 11223065
references.citation: Nat Biotechnol. 2007 Nov;25(11):1290-7
references.pmid: 17934452
references.citation: J Mol Biol. 1997 Jul 4;270(1):26-35
references.pmid: 9231898
references.citation: Curr Opin Immunol. 2002 Dec;14(6):698-704
references.pmid: 12413518
references.citation: Transplantation. 1999 Dec 15;68(11):1632-7
references.pmid: 10609938
references.citation: Trends Biotechnol. 2004 May;22(5):238-44
references.pmid: 15109810
references.citation: Science. 2000 Oct 6;290(5489):84-9
references.pmid: 11021804
references.citation: Immunol Today. 2000 Aug;21(8):391-7
references.pmid: 10916142
references.citation: J Biol Chem. 2001 Mar 2;276(9):6591-604
references.pmid: 11096108
references.citation: Annu Rev Immunol. 1999;17:875-904
references.pmid: 10358776
references.citation: J Allergy Clin Immunol. 2000 Aug;106(2):337-48
references.pmid: 10932079
references.citation: J Allergy Clin Immunol. 2005 Mar;115(3):S376-85
references.pmid: 15753882
references.citation: Biochem J. 1989 Apr 15;259(2):347-53
references.pmid: 2524188
references.citation: J Immunol. 1997 Feb 1;158(3):1438-45
references.pmid: 9013989
references.citation: Science. 2009 Mar 20;323(5921):1610-4
references.pmid: 19299620
references.citation: J Immunol Methods. 2002 May 1;263(1-2):133-47
references.pmid: 12009210
references.citation: J Immunol. 1989 Oct 15;143(8):2595-601
references.pmid: 2507634
references.citation: J Allergy Clin Immunol. 2008 Feb;121(2):320-5
references.pmid: 18086492
references.citation: J Mol Biol. 2004 Jul 23;340(5):1073-93
references.pmid: 15236968
references.citation: Expert Opin Ther Targets. 2005 Feb;9(1):169-90
references.pmid: 15757489
references.citation: Nat Med. 2002 May;8(5):518-21
references.pmid: 11984598
references.citation: Nat Biotechnol. 2005 Sep;23(9):1126-36
references.pmid: 16151406
references.citation: J Biol Chem. 1991 Jun 15;266(17):11245-51
references.pmid: 1710221
references.citation: J Mol Biol. 2004 Apr 23;338(2):299-310
references.pmid: 15066433
references.citation: Science. 2008 Aug 15;321(5891):974-7
references.pmid: 18703743
references.citation: J Mol Biol. 2007 Feb 23;366(3):815-29
references.pmid: 17196977
references.citation: J Biol Chem. 2007 Sep 21;282(38):27659-65
references.pmid: 17644522
references.citation: Curr Opin Mol Ther. 2009 Feb;11(1):22-30
references.pmid: 19169956
references.citation: Proc Natl Acad Sci U S A. 1992 May 15;89(10):4285-9
references.pmid: 1350088
references.citation: J Clin Invest. 1995 Feb;95(2):577-85
references.pmid: 7860741
references.citation: Nat Biotechnol. 1998 Jul;16(7):677-81
references.pmid: 9661204
references.citation: Biomol Eng. 2001 Sep;18(2):31-40
references.pmid: 11535414
references.citation: Curr Opin Immunol. 1999 Dec;11(6):621-5
references.pmid: 10631545
references.citation: Adv Immunol. 2007;93:63-119
references.pmid: 17383539
references.citation: Allergy. 2004 Jul;59(7):772-80
references.pmid: 15180766
references.citation: Nat Med. 2005 Apr;11(4):446-9
references.pmid: 15793580
references.citation: J Allergy Clin Immunol. 2005 Mar;115(3):459-65
references.pmid: 15753888
references.citation: J Allergy Clin Immunol. 2004 Sep;114(3):527-30
references.pmid: 15356552
references.citation: Proc Natl Acad Sci U S A. 2010 Jan 12;107(2):604-9
references.pmid: 20080725
references.citation: Immunity. 1995 Nov;3(5):635-46
references.pmid: 7584153
references.citation: Acta Pharmacol Sin. 2005 Jun;26(6):649-58
references.pmid: 15916729