Epitope mapping from real time kinetic studies – Role of cross-linked disulphides and incidental interacting regions in affinity measurements: Study with human chorionic gonadotropin and monoclonal antibodies

Journal of biosciences

P Tamil Selvi¹  Nonavinakere Seetharam Srilatha¹  Gundlupet Satyanarayana Murthy1¹ 

[1] Department of Molecular Reproduction, Development and Genetics, Indian Institute of Science, Bangalore 560 012, India$$

关键词: Association constant;    epitope mapping;    incidental interacting regions;    kinetics;    monoclonal antibody;   

DOI  :  

来源: Indian Academy of Sciences

【 摘 要 】

Real time kinetic studies were used to map conformational epitopes in human chorionic gonadotropin (hCG) for two monoclonal antibodies (MAbs). The epitopes were identified in the regions (𝛼5–14 and 𝛼55–62). The association rate constant (𝑘+1) was found to be altered by chemical modification of hCG, and the ionic strength of the reaction medium. Based on these changes, we propose the presence of additional interactions away from the epitope-paratope region in the hCG-MAb reaction. We have identified such incidental interacting regions (IIRs) in hCG to be the loop region 𝛼35–47 and 𝛼60–84. The IIRs contribute significantly towards the 𝐾𝐴 of the interaction. Therefore, in a macromolecular interaction of hCG and its MAb, 𝐾𝐴 is determined not only by epitopeparatope interaction but also by the interaction of the nonepitopic-nonparatopic IIRs. However, the specificity of the interaction resides exclusively with the epitope-paratope pair.

【 授权许可】

Unknown