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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.
Artículo en Chino | 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.
Asunto(s)
Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Virología / Sangre / Proteínas Recombinantes / Anticuerpos Anti-VIH / Infecciones por VIH / Sensibilidad y Especificidad / VIH-1 / Clasificación / Renaturación de Proteína / Alergia e Inmunología Tipo de estudio: Estudio diagnóstico Límite: Humanos Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2010 Tipo del documento: Artículo

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Texto completo: Disponible Índice: WPRIM (Pacífico Occidental) Asunto principal: Virología / Sangre / Proteínas Recombinantes / Anticuerpos Anti-VIH / Infecciones por VIH / Sensibilidad y Especificidad / VIH-1 / Clasificación / Renaturación de Proteína / Alergia e Inmunología Tipo de estudio: Estudio diagnóstico Límite: Humanos Idioma: Chino Revista: Chinese Journal of Biotechnology Año: 2010 Tipo del documento: Artículo