Expression, purification and renaturation of Pol P51 antigen of HIV-1 strain CN54 and its application in antibody detection / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 201-206, 2010.
Artigo
em Chinês
| WPRIM
| ID: wpr-336241
ABSTRACT
To obtain the pure and soluble P51 antigen of HIV-1 strain CN54, we transformed the Escherichia. coli strain BL21 codonplus-RIL with recombinant plasmid pTHioHisA51 which carries a gene encoding the Polymerase (Pol) P51 antigen of HIV-1 CN54 formerly, and induced protein expression by IPTG. We purified the recombinant protein with Chelating Sepharose FF-Ni and DEAE-Sepharose FF column chromatography, then renatured the recombinant protein by dialyzation. Purified protein was identified by Western blotting. We labeled and coated antigen P51 in a dual-antigen sandwich system, and tested it with serum samples from HIV-infected individuals. The results showed that P51 was expressed as inclusion body, and represented about 50% of total cellular protein. After purification and renaturation, the purity of P51 was up to 95%. Western blotting and sandwich ELISA demonstrated that recombinant P51 had good anti-HIV antibody specificity and sensitivity. The results suggested that recombinant HIV-1 P51 can be prepared as diagnostic reagent, and provides valuable support for HIV-1 detection and vaccine research.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Virologia
/
Sangue
/
Proteínas Recombinantes
/
Anticorpos Anti-HIV
/
Infecções por HIV
/
Sensibilidade e Especificidade
/
HIV-1
/
Classificação
/
Renaturação Proteica
/
Alergia e Imunologia
Tipo de estudo:
Estudo diagnóstico
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Biotechnology
Ano de publicação:
2010
Tipo de documento:
Artigo
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