Isotype choice for chimeric antibodies affects binding properties.

M M Morelock; R Rothlein; S M Bright; M K Robinson; E T Graham; J P Sabo; R Owens; D J King; S H Norris; D S Scher

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PubMed · 7909805

Isotype choice for chimeric antibodies affects binding properties.

M M Morelock·R Rothlein·S M Bright·M K Robinson·E T Graham·J P Sabo·R Owens·D J King·S H Norris·D S Scher

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Abstract

Construction of a series of chimeric antibodies (murine variable region and human constant region) derived from the murine antibody BIRR1, which recognizes intercellular adhesion molecule 1 (ICAM-1), has revealed differences in the relative binding abilities of the chimeric antibody to antigen. The chimeric antibodies show a ranking of their ability to compete with BIRR1 for antigen on the surface of cells with the order BIRR1 = cIgG1 (100%) > cIgG4 (30%) > cIgG2 (10%) as demonstrated by solid-phase competitive enzyme-linked immunosorbent assay. Papain digestion yielded Fab fragments that were purified to homogeneity. Competitive enzyme-linked immunosorbent assay showed that the chimeric and murine Fab binding constants were equivalent. A solution-phase binding assay (analyzed by size exclusion high performance liquid chromatography) between the intact mAbs and recombinant soluble ICAM-1 further established that the binding constants involving the Fab arms of the two antibodies were equivalent. In summary, the murine and chimeric anti-ICAM-1 antibodies bind cellular ICAM-1 with equivalent affinities but with differing avidities.

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Animals Antibodies, Monoclonal Base Sequence Binding Sites, Antibody CHO Cells Cell Adhesion Molecules Cricetinae Enzyme-Linked Immunosorbent Assay Humans Immunoglobulin Fab Fragments Immunoglobulin Isotypes Intercellular Adhesion Molecule-1 Mice Molecular Sequence Data Oligodeoxyribonucleotides Recombinant Fusion Proteins Journal Article

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M M Morelock, R Rothlein, S M Bright, M K Robinson, E T Graham, J P Sabo, R Owens, D J King, S H Norris, D S Scher. 1994-04-29. Isotype choice for chimeric antibodies affects binding properties.. https://pubmed.ncbi.nlm.nih.gov/7909805/

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