Preparation and identification of anti-HIV-1 gp120 monoclonal antibody
- VernacularTitle:抗HIV-1 gp120单克隆抗体的制备及鉴定
- Author:
Yongchen ZHANG
;
Jian WEN
;
Liang ZHANG
;
Lei ZHAO
- Publication Type:Journal Article
- Keywords:
HIV-1 gp120 antigen;
prokaryotic expression;
monoclonal antibody;
hybridoma cell
- From:
Chinese Journal of Biochemical Pharmaceutics
2010;31(1):26-28,31
- CountryChina
- Language:Chinese
-
Abstract:
Purpose To Prepare anti- HIV-1 gp120 monoclonal antibodies and to identify the specificity of antibodies in order to provide technique for preparing HIV remedial antibodies. Methods The gene fragment of HIV-1 gp120 was connected to PEGX-4T-2 prokaryotic expressing vector. The vector was cut by enzyme. GST-HIV protein was expressed by E. coli XL1-blue. The BALB/c mice were immunized with purified GST-HIV antigen, and then the fusion of mice spleen cell and myeloma cell SP2/0 was executed as the routine cell-fusion technique. Positive cells were screened by Indirect ELISA. Immuno-blotting assay and Western blot identified the specificity of antibodies. Results External gene section from the recombinant plasmid by sequencing showed the same size of HIV-1 gp120 gene sequences. An external expressed protein band of 32 KD was obtained after purified protein SDS-PAGE electrophoresis. It indicated that six strains of hybridoma cells secreting special monoclone antibodies had been obtained. ELISA results showed that six strains monoclonal antibodies only reacted to HIV-1 gp120 antigen. Western blot results showed that a band with molecular weight 32 kDa was obtained, which could interact with HIV-1 gp120 monoclonal antibody. Conclusion Six strains of hybridoma cells secreting special monoclone antibodies had been obtained. The prepared monoclonal antibodies have established a basis for HIV remedial antibody.
