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
em Zh
| WPRIM
| ID: wpr-354755
Biblioteca responsável:
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 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.
Texto completo:
1
Índice:
WPRIM
Assunto principal:
Peptídeos
/
Virologia
/
Sangue
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Ensaio de Imunoadsorção Enzimática
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Infecções por HIV
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Linhagem Celular
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HIV-1
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Alergia e Imunologia
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Produtos do Gene vpr do Vírus da Imunodeficiência Humana
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Genética
Tipo de estudo:
Diagnostic_studies
Limite:
Animals
/
Humans
Idioma:
Zh
Revista:
Chinese Journal of Virology
Ano de publicação:
2012
Tipo de documento:
Article