Comparison of recombinant human immunodeficiency virus gag precursor and gagenv fusion proteins and a synthetic env peptide as diagnostic reagents

✍ Scribed by R.L. Shoeman; D. Young; R. Pottathil; J. Victor; R.R. Conroy; R.M. Crowl; T. Coleman; E. Heimer; C.-Y. Lai; K. Ganguly; E.P. Reddy; A.M. Skalka; P.R. Pine; F.R. Khan; H. Weissbach

• Publisher: Elsevier Science
• Year: 1987
• Tongue: English
• Weight: 1006 KB
• Volume: 161
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(87)90465-9

✦ Synopsis

Diagnostic reagents for detection of human immunodeficiency virus (HIV) exposure with improved reliability may be provided by viral encoded proteins produced by recombinant DNA techniques or by synthetic peptides corresponding to appropriate viral epitopes. We have expressed at high levels in E. coli a gag gene segment corresponding to -97% of the ~55 gag precursor protein, as well as a novel gag/env fusion protein that contains antigenic determinants in common with gag ~24, env gp4 1, and env gpl20. The gag and gag/env proteins were purified from insoluble inclusion bodies by sequential extraction with increasing concentrations of urea. These components were tested for reactivity with antisera to HIV proteins and peptides. We have also chemically synthesized a peptide corresponding to env residues 578-608, representing a portion of env gp4 1. The final preparation of gag and gag/env proteins in 8 M urea reacted with sheep anti-HTLV-III p24 gag antibodies and acquired immune deficiency syndrome (AIDS) patient sera. The gag/env fusion protein also reacted with rabbit anti-HIV env 500-5 1 I peptide antibody. Both recombinant proteins and the env peptide were suitable as reagents for evaluation of serum samples by enzyme-linked immunosorbent assay (ELISA). Results of ELISA assays utilizing the recombinant viral proteins and synthetic peptide were in good agreement with results obtained using disrupted virus as antigen in ELISA assays and immunoblotting.