用抗Der f 2小鼠单克隆抗体分析Der f 2的人IgE表位

Analysis of human IgE epitope of Der f 2 with anti-Der f 2 mouse monoclonal antibodies.

作者信息

Nishiyama C, Hatanaka H, Ichikawa S, Fukada M, Akagawa-Chihara M, Yuuki T, Yokota T, Inagaki F, Okumura Y

机构信息

Bioscience Research and Development Laboratory, Asahi Breweries Ltd, Ibaraki, Japan.

出版信息

Mol Immunol. 1999 Jan;36(1):53-60. doi: 10.1016/s0161-5890(98)00112-6.

DOI:10.1016/s0161-5890(98)00112-6

PMID:10369420

Abstract

Der f 2 is one of the major mite allergens recognized by human IgE antibodies of allergic patients. Using five anti-Der f 2 mouse monoclonal antibodies, human IgE epitopes of Der f 2 were analyzed. Among them, two monoclonal antibodies 15E11 and 13A4 inhibited the binding between Der f 2 and human IgE antibodies. To determine major IgE epitopes of Der f 2, epitopes for the monoclonal IgG antibodies were analyzed using 43 single site Der f 2 mutants constructed by site-directed mutagenesis. Binding ability of 13A4 and 15E11 was decreased by the amino acid replacement around the C-terminus, and around 73rd, respectively. These results suggest that the C-terminal portion and the central portion around 73rd of Der f 2 were recognized by human IgE antibodies as major epitopes. The location of the putative IgE epitopes on 3-D structure of Der f 2 is also discussed.

摘要

Der f 2是过敏性患者的人IgE抗体识别的主要螨过敏原之一。使用五种抗Der f 2小鼠单克隆抗体，对Der f 2的人IgE表位进行了分析。其中，两种单克隆抗体15E11和13A4抑制了Der f 2与人IgE抗体之间的结合。为了确定Der f 2的主要IgE表位，使用通过定点诱变构建的43个单位点Der f 2突变体分析了单克隆IgG抗体的表位。13A4和15E11的结合能力分别因C末端周围和第73位左右的氨基酸替换而降低。这些结果表明，Der f 2的C末端部分和第73位左右的中心部分被人IgE抗体识别为主要表位。还讨论了推定的IgE表位在Der f 2三维结构上的位置。