Recombinant HIV-1 p24 protein: cloning, expression, purification and use in the development of ELISA kits

• Published in: Current science (Bangalore) Vol. 91; no. 7; pp. 913 - 917
• Main Authors: Bhardwaj, Devesh, Bhatt, Seema, Khamar, Bakulesh M., Modi, Rajiv I., Ghosh, Prasanta K.
• Format: Journal Article
• Language: English
• Published: Current Science Association 10.10.2006
• Subjects: AIDS; Antibodies; Antigens; B lymphocytes; Enzyme linked immunosorbent assay; Escherichia coli; Gels; HIV; HIV 1; HIV 2; Human immunodeficiency virus 1; Human immunodeficiency virus 2; Recombinant proteins
• ISSN: 0011-3891

A simple method for purification of recombinant p24 (rp24) of human immunodeficiency virus type 1 (HIV-1) from Escherchia coli is described. The protein was over-expressed in E. coli (BL21λDE3) cells to levels of ∼30% total cell protein in the soluble fraction. The recombinant protein was extracted and purified to near homogeneity by simple ion-exchange steps followed by gel filtration. The recombinant protein in combination with synthetic peptides from immuno-dominant stretches derived from the 'envelope' proteins of HIV-1 and HIV-2 was used for development of an ELISA kit, ELIK HIV 1 & 2 for detection of antibodies against the virus in human serum and/or plasma. The kit was found to have 99.63% sensitivity and 99.88% specificity in a performance study using more than 2100 well-defined samples.