Identification of binding epitope of a monoclonal antibody (Z12) against human TNF-α using computer modeling and deletion mutant technique

作     者：ZHANG Wei FENG Jiannan SHEN Beifen 

作者机构：Beijing Basic Medical Institute Beijing 100850 China Beijing Basic Medical Institute Beijing 100850 China Beijing Basic Medical Institute Beijing 100850 China 

出 版 物：《Science China(Life Sciences)》 (中国科学（生命科学英文版）)

年 卷 期：2004年第47卷第3期

页      面：279-286页

核心收录：

学科分类：07[理学] 08[工学] 

基　　金：This study has applied for a native inventing patent whose number(03104870.6) 

主　　题：human tumor necrosis factor-α, Fv, antigenic epitope, homology-based modeling, deletion. 

摘      要：The genes of the heavy and light chain variable region (VH, VL) of Z12 antibody against hTNF-a were cloned, and according to the translated sequence of amino acids, the spa-tial structures of VH and VL domains were modeled by using homology-based modeling method, followed by constructing the whole three-dimensional structure of Fv fragment. The complex model of Fv interacting with hTNF-a was gained with computer-guided molecular docking method, based on which, it was predicted that the epitope recognized by Z12 was from 141 to 146 of hTNF-a. hTNF-a molecule was divided into two fragments of N-terminal region from 1 to 91 and C-terminal region from 92 to 157 with prokaryotic expression. The measured results suggested that the antigenic epitope recognized by Z12 antibody was located in the C-terminal region 92-157 of hTNF-a, proving the predicted result reliable preliminarily. Further experimental results showed that after hTNF- 141-146 residues were deleted, Z12 antibody almost lost the ability to recognize the mutant, suggesting that the amino acid residues from 141 to 146 of hTNF-a were specially recognized by Z12 antibody.