Development and identification of polyclonal antibodies against HIV-1 Vpr-derived polypeptides / 病毒学报
Chinese Journal of Virology
; (6): 151-157, 2012.
Article
в Zh
| WPRIM
| ID: wpr-354755
Ответственная библиотека:
WPRO
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 1100000 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.
Полный текст:
1
База данных:
WPRIM
Основная тема:
Peptides
/
Virology
/
Blood
/
Enzyme-Linked Immunosorbent Assay
/
HIV Infections
/
Cell Line
/
HIV-1
/
Allergy and Immunology
/
Vpr Gene Products, Human Immunodeficiency Virus
/
Genetics
Тип исследования:
Diagnostic_studies
Пределы темы:
Animals
/
Humans
Язык:
Zh
Журнал:
Chinese Journal of Virology
Год:
2012
Тип:
Article