针对人白细胞介素-5的大鼠单克隆Fab片段的结构与人性化改造

Structure and humanization of a rat monoclonal Fab to human interleukin-5.

作者信息

Cook W J, Walter L J, Murgolo N J, Chou C C, Petro M, Zavodny P J, Narula S K, Ramanathan L, Trotta P P, Nagabhushan T L

机构信息

Department of Pathology, University of Alabama at Birmingham 35233-6823, USA.

出版信息

Protein Eng. 1996 Jul;9(7):623-8. doi: 10.1093/protein/9.7.623.

DOI:10.1093/protein/9.7.623

PMID:8844835

Abstract

The X-ray crystal structure of a rat monoclonal Fab JES1-39D10, raised against recombinant human interleukin-5, has been determined with the use of molecular replacement techniques and refined at 2.7 A resolution by simulated annealing. The overall structure is similar to a murine Fab HyHEL-10 that is specific for hen egg white lysozyme. An interesting feature of the structure is the presence of leucine residues to support the H1 complementarity-determining region (CDR) loop. To our knowledge this is the first Fab crystal structure containing this unusual H1 loop support pattern. The activity of three humanized versions of 39D10 is explained by analysis of Fv interface residues and H1 support patterns of 39D10 and the human template HIL.

摘要

利用分子置换技术确定了针对重组人白细胞介素-5产生的大鼠单克隆Fab JES1-39D10的X射线晶体结构，并通过模拟退火在2.7埃分辨率下进行了精修。其整体结构类似于对鸡蛋清溶菌酶具有特异性的鼠源Fab HyHEL-10。该结构的一个有趣特征是存在亮氨酸残基以支撑H1互补决定区（CDR）环。据我们所知，这是首个包含这种不寻常H1环支撑模式的Fab晶体结构。通过对39D10的Fv界面残基以及39D10和人源模板HIL的H1支撑模式进行分析，解释了39D10的三种人源化形式的活性。