The His‐probe method: Effects of histidine residues introduced into the complementarity‐determining regions of antibodies on antigen‐antibody interactions at different pH values

FEBS Letters

Fujio, Hajime⁴  Kurosawa, Yoshikazu¹  Ito, Wataru¹  Arata, Yoji³  Sakato, Nobuo⁴  Yutani, Katsuhide² 

[1] Institute for Comprehensive Medical Science, Fujita Health University, Toyoake, Aichi 470-11, Japan;Institute for Protein Research, Osaka University, Suita, Osaka 565, Japan;Faculty of Pharmaceutical Sciences, University of Tokyo, Hongo, Tokyo 113, Japan;Department of Immunology, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka 565, Japan

关键词: Binding constants;    Site-directed mutagenesis;    Ab;    antibody;    Ag;    antigen;    CDR;    complementarity-determining region;    HEL;    hen egg-white lysozyme;    ELISA;    enzyme-linked immunosorbent assay;    CD;    circular dichroism;   

DOI  :  10.1016/0014-5793(92)80745-3

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We examined the effects or histidine residues that were artificially introduced into complementarity-determining regions of antibodies on antigen-antibody Interactions at different pH values. Using a monoclonal antibody specific ror hen egg-white lysozyme and three mutant antibodies that contained a histidine residue. we measured binding constants for antibodies and lysozyme at different pH values (PH 5–8). No gross conformational changes were evident over this range of pH vulues, as determined by analysis of the spectra of circular dichroism. Since the charge on a histidine residue is the most likely factor that can vary over this range of pH values, differences on pH-dependent antigen-binding patterns observed between the wild-type and mutant antibodies should be due mainly to the effects of the charges on the histidine residues. The three mutant antibodies showed different and characteristic patterns of pH-dependent binding to lysozyme, which depended on the location of the artificially introduced histidine residues.

【 授权许可】

Unknown