Gilbert M, Kirihara J, Mills J; International Conference on AIDS.
Int Conf AIDS. 1990 Jun 20-23; 6: 320 (abstract no. 1027).
University of California, San Francisco, California, USA
OBJECTIVES: A test that can measure gp120 concentration would be useful in laboratory and clinical studies with HIV. METHODS: Recombinant, soluble CD4 (rsCD4) immobilized in microtiter trays was used to capture gp120, which was then detected with a three-layer detector-amplification system consisting of polyclonal sheep anti-gp120, biotinylated rabbit anti-sheep IgG, and avidin-alkaline phosphatase. Samples were incubated with p-nitrophenyl-phosphate and absorbance read at three and thirty-three minutes. RESULTS: The assay showed a linear relationship between optical density and reference gp120 (recombinant, full-length, glycosylated gp120) concentration from 60 to 6000 pg/ml; the precision of the assay varied with the sample size, and ranging from +/- 50% with concentrations below 200 pg, to +/- 10% at concentrations above 200 pg/ml. In a group of replicate coded samples containing 60 pg/ml (approximately 10(7) molecules of gp120), the assay correctly identified the samples as containing gp120 90% of the time, with no false positives recorded in blank samples. Recombinant gp120 prepared in another cell culture system showed a binding coefficient which was 13-fold less than the reference gp120. With the assay system we were able to detect gp120 in HIV-1 suspensions prepared from cultured lymphoblastoid cells, and in the sera of HIV-1 infected patients. CONCLUSIONS: This gp120 ELISA should be useful for studying the biology of gp-120 in HIV-1 infection.
Publication Types:
Keywords:
- Acquired Immunodeficiency Syndrome
- Aluminum Hydroxide
- Animals
- Antigens, CD4
- Biological Assay
- Carbonates
- Enzyme-Linked Immunosorbent Assay
- Glycoproteins
- HIV Envelope Protein gp120
- HIV Infections
- HIV Seropositivity
- HIV-1
- Humans
- Link
- Rabbits
- Recombinant Proteins
- SK&F 106528
- enzymology
- immunology
Other ID:
UI: 102196873
From Meeting Abstracts