Cleavage and purification of prokaryotically expressed HIV gag and env fusion proteins for detection of HIV antibodies in the ELISA

Virology. 1991 Feb;180(2):811-3. doi: 10.1016/0042-6822(91)90097-u.

Abstract

Parts of the gag p24 and the gp41 transmembrane protein of the human immunodeficiency virus HIV-1 were expressed as fusion proteins in Escherichia coli, using an expression vector carrying aa 1-375 of the lac-Z gene linked to the recognition sequence for the blood coagulation factor Xa. Fusion proteins were cleaved into the bacterial and viral portion and the viral polypeptide was purified by a molecular sieve column. The purified viral antigens were tested with 288 human sera in the enzyme-linked immunosorbent assay (ELISA) technique. Comparison with commercially available tests showed comparable sensitivity and a higher specificity of the gag/env-ELISA for borderline reactive sera.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, Gel
  • Cloning, Molecular
  • Enzyme-Linked Immunosorbent Assay
  • Escherichia coli / genetics
  • Gene Products, env / immunology*
  • Gene Products, env / isolation & purification
  • Gene Products, gag / immunology*
  • Gene Products, gag / isolation & purification
  • HIV Antibodies / analysis*
  • HIV Seropositivity*
  • HIV-1 / immunology*
  • Humans
  • Viral Fusion Proteins / immunology*
  • Viral Fusion Proteins / isolation & purification

Substances

  • Gene Products, env
  • Gene Products, gag
  • HIV Antibodies
  • Viral Fusion Proteins