The molecular and biochemical characterization of mutant monoclonal antibodies with increased antigen binding.
Mutant mAb with increased Ag binding were generated from a hybridoma cell line, 36-65, that secretes an IgG1,kappa anti-p-azophenylarsonate-(Ars) specific antibody. The mutant antibodies were identified using an Ars-specific ELISA and sib selection so that approximately 10(6) cells could be analyzed. The ELISA used as Ag a low ratio of Ars coupled to BSA and was set up so that only those antibodies that had higher binding than the parent would be detected. Seven mutant producing cell lines were isolated from five independent clones of 36-65. The mutant antibodies bind Ag 20 to more than 200-fold better than the parent and have wild type V region sequences. All have C region mutations that result in an increased avidity. At least five different genetic events are responsible for the C region mutations.
French
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Department of Pathology, State University of New York, Stony Brook 11794-8691.
Pollock
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Aguila
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Scharff
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D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics* / Q000276:immunology; D000915:Antibody Affinity / Q000235:genetics; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007127:Immunoglobulin Constant Regions / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009154:Mutation; D011061:Poly A / Q000235:genetics; D011108:Polymers / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D010132:p-Azobenzenearsonate / Q000276:immunology
D000911:Antibodies, Monoclonal; D007127:Immunoglobulin Constant Regions; D007143:Immunoglobulin Heavy Chains; D011108:Polymers; D012333:RNA, Messenger; D011061:Poly A; D010132:p-Azobenzenearsonate