A sensitive radioimmunoprecipitation assay for the detection and quantitation of antibodies to the envelope glycoprotein gp120 of the human immunodeficiency virus (HIV-1).
A radioimmunoprecipitation (RIP) assay was developed to detect antibodies to the envelope glycoprotein gp120 of the human immunodeficiency virus (HIV-1). The assay, which utilized recombinant gp120 (rgp120), was quantitative, reproducible, and specific for antibodies to rgp120 or antibodies to native gp120 resulting from natural infection with HIV. Polyethylene glycol-8000 (PEG), used in the assay at a final concentration of 10% to precipitate immune complexes, was demonstrated to be effective in titering sera from different animal species. Provided samples were diluted at least 1:100, antibody titers could be determined either by the classical dilution method or by interpolation from a calibration curve prepared with a positive serum. The humoral response of animals immunized with rgp120 was monitored and a positive correlation was found between titers determined in the RIP assay and the ability of the sera to neutralize. In addition, RIP titers of HIV-positive human sera correlated very well with reactivity obtained in a commercial HIV immunoblot assay. The assay has the advantage of quantitation, fast turnaround time, and versatility.
Lucas
C
C
Department of Medicinal and Analytical Chemistry, Genentech Inc., SouthSan Francisco, CA 94080.
Peterson
M L
ML
Bennett
G L
GL
Frie
S W
SW
Berman
P W
PW
Chen
A B
AB
D000818:Animals; D002138:Calibration; D006041:Goats; D006168:Guinea Pigs; D015483:HIV Antibodies / Q000032:analysis*; D015699:HIV Envelope Protein gp120 / Q000276:immunology*; D006801:Humans; D015151:Immunoblotting; D007457:Iodine Radioisotopes; D009500:Neutralization Tests; D002679:Pan troglodytes; D010215:Papio; D011233:Precipitin Tests; D011817:Rabbits; D011863:Radioimmunoassay; D011994:Recombinant Proteins / Q000276:immunology
D015483:HIV Antibodies; D015699:HIV Envelope Protein gp120; D007457:Iodine Radioisotopes; D011994:Recombinant Proteins