Preparation and characterization of a monoclonal antibody against HIV-1 inhibitor N-SRCR
- VernacularTitle:HIV-1抑制蛋白N-SRCR单克隆抗体的制备与鉴定
- Author:
Chao NIE
;
Zhong LIU
;
Xin HE
;
Zhiwei WU
- Publication Type:Journal Article
- Keywords:
Scavenger veceptor cysteine-rich domain;
Monoclonal antibody;
Human immunodefficiency virus
- From:
Journal of Medical Postgraduates
2004;0(01):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective Protein N-SRCR derived from salivary agglutinin (SAG) inhibits HIV-1 infection.An N-SRCR monoclonal antibody was prepared for the study of the interaction between N-SRCR and HIV-1 envelop glycoprotein (gp120).Methods The purified recombinant N-SRCR expressed by 293 cells was used to immunize four weeks old BALB/c mice.After the final boost,the mouse spleen cells were isolated and fused with mouse myeloma cell line SP2/0-Ag-14,and the resulting hybridomas were screened for the production of N-SRCR-specific antibodies by ELISA assay.The monoclonal antibody against N-SRCR was purified by HiTrap Protein G kit,the purity determined by SDS-PAGE and the antibody titers by ELISA.The antibody specificity.was charqacterized by western blotting.Results A strain of hybridoma cell clones stably secreting N-SRCR antibody,named 1D6,was obtained.The high purity of the IgG was demonstrated by SDS-PAGE,and the ELISA titers of 1D6 was more than 100?25.Conclusion A monoclonal antibody against N-SRCR was successfully prepared,which laid the ground for further studies on the biological function of N-SRCR and the interactions between SRCR domains and HIV-1 Env gp120.
