Exploration on a new routine for preparing rabies laboratory diagnostic monoclonal antibodies
10.3760/cma.j.issn.1003-9279.2014.03.026
- VernacularTitle:一种狂犬病实验室诊断用单克隆抗体制备新路线的探索
- Author:
Xinjun LV
1
;
Guodong LIANG
;
Qing TANG
Author Information
1. 中国疾病预防控制中心病毒病预防控制所
- Keywords:
Rabies virus;
Nucleoproteins;
Peptides;
Antibody/diagnosis
- From:
Chinese Journal of Experimental and Clinical Virology
2014;28(3):236-238
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the possibility of preparing rabies laboratory diagnostic monoclonal antibodies with synthetic peptides as immunogen.Methods A synthetic peptide located at residues 355 to 369 of rabies virus CVS-11 nucleoprotein was conjugated to the Keyhole Limpe hemocyanin (KLH) macromolecule and used to immunize BALB/c mice to prepare hybridoma cell lines by classical hybridoma technology.Indirect enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent assay (IFA) were used to screen and identify the hybridoma cell lines.Results A positive hybridoma cell line named 2B1D11 was obtained after screening the anti-peptide antibodies in supernatants of hybridoma cell lines by indirect ELISA and IFA.Monoclonal antibody in ascites of BALB/c mice inoculated with 2B1D11 was purified and used successfully to detect the rabies virus in canine brain tissues and BHK-21 cells by IFA.Conclusion It is possible to prepare rabies laboratory diagnostic monoclonal antibodies with synthetic peptides as immunogen.
