Development and identification of polyclonal antibodies against HIV-1 Vpr-derived polypeptides.
- Author:
Jun SUN
1
;
Zhe-Feng MENG
;
Jian-Qing XU
;
Xiao-Yan ZHANG
;
Jian-Xin LV
Author Information
1. Wenzhou Medical College, Wenzhou 325000, China. sunjunapple@163.com
- Publication Type:Journal Article
- MeSH:
Animals;
Antibodies, Viral;
blood;
immunology;
Cell Line;
Enzyme-Linked Immunosorbent Assay;
HIV Infections;
blood;
immunology;
virology;
HIV-1;
genetics;
immunology;
Humans;
Peptides;
genetics;
immunology;
Rabbits;
vpr Gene Products, Human Immunodeficiency Virus;
genetics;
immunology
- From:
Chinese Journal of Virology
2012;28(2):151-157
- CountryChina
- Language:Chinese
-
Abstract:
To develop polyclonal antibodies against predicted B cell epitopes in HIV-1 accessory protein Vpr, the prepared consensus Vpr amino acid sequence was used to predict potential B cell epitopes by online softwares (ABCpred and Bcepred), the synthesized polypeptides of B-cell epitopes were subsequently conjugated with keyhole limpet hemocyanin (KLH) and then used to immunize rabbits. The antibody titers were determined by ELISA, and antibody specifity was analyzed by Western-Blotting and immunoprecipitation, respectively. Amino acid residues 3-19 (N) and 82-95 (C) of Vpr were predicted as the potential B cell epitopes. After inoculation of the conjugation of synthesized peptide to KLH, the antibody titers in rabbit sera against N and C peptides reached more than 1:100000 by ELISA. Western-Blotting analysis showed that the polyclonal antibodies reacted with both wild Vpr and fusion protein of GFP with Vpr, no matter Vpr was derived from HIV-1 B subtype or CRF07_BC recombinant form; Immunoprecipitation analysis showed similar reactions to Western-Blotting results. Two B cell epitopes of Vpr were successfully predicted by Bio-informatics methods and polyclonal antibodies against those peptides could be successfully prepared.
