一种针对在毕赤酵母中表达的登革4型病毒重组包膜蛋白的单克隆抗体的研制。

Development of a monoclonal antibody specific to a recombinant envelope protein from dengue virus type 4 expressed in Pichia pastoris.

作者信息

Pupo-Antúnez M, Rodriguez R, Alvarez M, Amin N, Rodriguez H, Otero A, Guzmán G

机构信息

Virology Department, Tropical Medicine Institute, Pedro Kouri, Havana City, Cuba.

出版信息

Hybridoma. 2001 Feb;20(1):35-41. doi: 10.1089/027245701300060409.

DOI:10.1089/027245701300060409

PMID:11289225

Abstract

A mouse monoclonal antibody (MAb, 4B6) was able to recognize dengue virus type 4 envelope (E) protein both as a recombinant protein in Pichia pastoris and when it was present in infected brains of suckling mice. 4B6 was characterized by enzyme-linked immunoadsorbent assay (ELISA), hemaglutination inhibition, neutralization, and immunoblot. The MAb was isotyped as IgG2a. It was serotype 4 specific and it inhibited hemaglutination and neutralized homologous virus. It did not enhance infection of P338D1 cells by dengue type 4 virus strain H-241 strain. This MAb was reactive with recombinant E protein and dengue 4 virus, as revealed by Western blot. In vivo, MAb 4B6 conferred passive protection in mice challenged with homologous virus. Currently, this MAb is being used to purify recombinant E protein for further studies.

摘要

一种小鼠单克隆抗体（MAb，4B6）能够识别4型登革病毒包膜（E）蛋白，无论是在毕赤酵母中作为重组蛋白，还是存在于乳鼠受感染的脑中时。通过酶联免疫吸附测定（ELISA）、血凝抑制、中和及免疫印迹对4B6进行了表征。该单克隆抗体的亚型为IgG2a。它具有血清型4特异性，能抑制血凝并中和同源病毒。它不会增强4型登革病毒H - 241株对P338D1细胞的感染。如免疫印迹所示，该单克隆抗体与重组E蛋白和登革4病毒反应。在体内，单克隆抗体4B6在用同源病毒攻击的小鼠中提供了被动保护。目前，该单克隆抗体正用于纯化重组E蛋白以进行进一步研究。