Cleavage and purification of prokaryotically expressed HIV gag and env fusion proteins for detection of HIV antibodies in the ELISA.
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.
Ellinger
S
S
Institut für Klinische und Molekulare Virologie, Universität Erlangen-Nürnberg, Federal Republic of Germany.
Mach
M
M
Korn
K
K
Jahn
G
G
D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics; D015686:Gene Products, env / Q000276:immunology* / Q000302:isolation & purification; D015683:Gene Products, gag / Q000276:immunology* / Q000302:isolation & purification; D015483:HIV Antibodies / Q000032:analysis*; D006679:HIV Seropositivity; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D014760:Viral Fusion Proteins / Q000276:immunology* / Q000302:isolation & purification
D015686:Gene Products, env; D015683:Gene Products, gag; D015483:HIV Antibodies; D014760:Viral Fusion Proteins