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.
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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