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Monoclonal antibodies to beta subunit of choriogonadotropin: Development and use in a sandwich ELISA pregnancy test

✍ Scribed by Rosana Gariglio; Emilena Toplikar; Amadeo Carlomagno; Federico Rojkin; Leonardo Knecher; Gustavo Capriotti; Dr. Luis Lorenzo

Publisher: John Wiley and Sons
Year: 1992
Tongue: English
Weight: 443 KB
Volume: 6
Category: Article
ISSN: 0887-8013
DOI: 10.1002/jcla.1860060505

No coin nor oath required. For personal study only.

✦ Synopsis

Balbic mice were immunized with beta subunit isolated and purified from crude human chorionic gonadotropin preparations. Spleen cells from the higher titered mouse were fused with Sp 2/0 myeloma cells.

Four specific secreting hybridomas were obtained. Specificity, affinity, and suitability of secreted antibodies for use in enzyme immunoassays were studied. Ascites of the selected hybridoma was raised; the antibody was purified by protein A-affinity chromatography and coupled to horseradish peroxidase. This conjugate was employed in a simultaneous sandwich enzyme immunoassay on microtiter plates sensitized with goat polyclonal antibody to measure the hormone.

The test has a sensitivity of 10 mlU/ml either on urine, serum, or plasma samples when read in a microplate reader. The results can also be evaluated by the naked eye, with a sensitivity of 20 mlU/ml. No cross reactivity was detected with other human gonadotropins. o 1992 Wiley-Liss, Inc.

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