An efficient route to human bispecific IgG.
Production of bispecific IgG (BsIgG) by coexpressing two different antibodies is inefficient due to unwanted pairings of the component heavy and light chains. To overcome this problem, heavy chains were remodeled for heterodimerization using engineered disulfide bonds in combination with previously identified "knobs-into-holes" mutations. One of the variants, S354C:T366W/Y349'C:T366'S:L368'A:Y407++ +'V, gave near quantitative (approximately 95%) heterodimerization. Light chain mispairing was circumvented by using an identical light chain for each arm of the BsIgG. Antibodies with identical light chains that bind to different antigens were identified from an scFv phage library with a very restricted light chain repertoire for the majority (50/55) of antigen pairs tested. A BsIgG capable of simultaneously binding to the human receptors HER3 and cMpI was prepared by coexpressing the common light chain and corresponding remodeled heavy chains followed by protein A chromatography. The engineered heavy chains retain their ability to support antibody-dependent cell-mediated cytotoxicity as demonstrated with an anti-HER2 antibody.
Merchant
A M
AM
Department of Molecular Oncology, Genentech Inc., South San Francisco, CA 94080, USA.
Zhu
Z
Z
Yuan
J Q
JQ
Goddard
A
A
Adams
C W
CW
Presta
L G
LG
Carter
P
P
D000918:Antibody Specificity / Q000276:immunology*; D000936:Antigen-Antibody Complex; D003602:Cytotoxicity, Immunologic; D019281:Dimerization; D004220:Disulfides; D066246:ErbB Receptors / Q000378:metabolism; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D008958:Models, Molecular; D009154:Mutation / Q000235:genetics; D009363:Neoplasm Proteins; D015202:Protein Engineering; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D020893:Receptor, ErbB-3; D018121:Receptors, Cytokine / Q000378:metabolism; D011971:Receptors, Immunologic / Q000378:metabolism*; D053628:Receptors, Thrombopoietin
D000936:Antigen-Antibody Complex; D004220:Disulfides; D007074:Immunoglobulin G; D009363:Neoplasm Proteins; D011518:Proto-Oncogene Proteins; D018121:Receptors, Cytokine; D011971:Receptors, Immunologic; D053628:Receptors, Thrombopoietin; C082218:MPL protein, human; D066246:ErbB Receptors; D020893:Receptor, ErbB-3