[Full book]R-ELISA: repeated use of antigen-coated plates for ELISA and its application for testing of antibodies to HIV and other pathogens

Post time 2024-4-30 00:39:55

journal: Biotechniques
Authors: Baunoch DA, Das P, Browning ME, Hari V.

Published date: March 1992

DOI: None found

PDF link: None found

Article link: R-ELISA: repeated use of antigen-coated plates for ELISA and its application for testing of antibodies to HIV and other pathogens - PubMed (nih.gov)

Remark: 1992 Mar;12(3):412-7.
Journal info:
Title(s):BioTechniquesPublication Start Year:1983Frequency:MonthlyCountry of Publication:EnglandPublisher:[Natick, MA : Eaton Pub. Co., c1983-Latest Publisher:London, UK : Future Science

Abstract:
In this paper we report on the evaluation of several procedures that allow for
the repeated use of an antigen-coated, enzyme-linked immunosorbent assay (ELISA)
plate for enzyme immunoassay (EIA). We have shown that antigen-coated ELISA
plates that were incubated once with an aqueous solution containing 8 M urea, 2%
sodium dodecyl sulfate and 2% mercaptoethanol, after an EIA, can be reused again
for EIA without loss of antigenic capacity. Thus, in this procedure, after an
EIA, the ELISA plates were washed once with the above solution and then in a
buffer containing 20 mM Tris-HCl, pH 7.5, 0.1% Tween 20 and 500 mM NaCl. This
washing protocol was shown to remove the primary antibody, enzyme-conjugated
secondary antibody and substrate without removing the antigen from the ELISA
plate microwells. Thus, an antigen-coated ELISA plate previously used for an
assay could be reused. We tested this repeat ELISA (R-ELISA) procedure on high
antigen-binding ELISA plates coated with two different plant virus proteins, a
synthetic peptide, the p25/24 gag and the gp120 proteins of the human
immuno-deficiency virus, or the staphylococcus enterotoxin protein. In each case
tested, the procedure allowed for the repeated use of the same antigen-coated
plates for EIA of the respective antibodies. This procedure should prove to be
particularly valuable for mass screening of samples tested for HIV and other
disease-causing agents.

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