Generation and characterization of specific monoclonal antibodies against monkeypox virus
10.3760/cma.j.issn.1003-9279.2017.02.015
- VernacularTitle: 猴痘病毒特异性单克隆抗体制备及鉴定
- Author:
Qianqian GUAN
1
;
Li ZHAO
;
Jiao REN
;
Panpan HUANG
;
Huijuan WANG
;
Yingzhu CHEN
;
Na ZHU
;
Wenjie TAN
;
Li RUAN
;
Houwen TIAN
Author Information
1. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China
- Publication Type:Journal Article
- Keywords:
Monkeypox virus;
A29 protein;
Polypeptide;
Monoclonal antibody
- From:
Chinese Journal of Experimental and Clinical Virology
2017;31(2):153-156
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To generate monkeypox virus specific monoclonal antibodies for further establishing monkeypox virus immunofluorescence assay.
Methods:Monkeypox virus A29 protein, vaccinia ortholog A27 protein and cowpox ortholog 162 protein were expressed in E. coli BL21 to screen antibodies. Synthetic monkeypox virus A2917 ~ 49 polypeptide was used to immune BALB/c mice. Monkeypox virus monoclonal antibodies were generated through fusion, cloning and screening techniques. Indirect ELISA was performed to test antibodies specificity and subtype.
Results:A29, A27 and 162 proteins were highly expressed in E. coli and detected by Western blot. The three his-tagged proteins were purified using His-Bind affinity chromatography column. The purity of the proteins was all more than 90%. And 8 strains monkeypox virus specific monoclonal antibodies were screened by the three purified proteins. Two mAbs of 8 were IgG3 subtype and the rest were IgG1 subtype.
Conclusions:Eight strains of monkeypox virus specific monoclonal antibodies were generated, they can be used to further establish monkeypox virus immune immunofluorescence assay.
