An automatic fluorescence micro-ELISA system for quantitative screening of hybridoma supernatants using a protein-A-β-galactosidase conjugate
- 1 January 1984
- journal article
- Published by Springer Nature in Journal of Molecular Histology
- Vol. 16 (1), 21-35
- https://doi.org/10.1007/bf01003433
Abstract
We report on a rapid micro-ELISA screening procedure for the detection of monoclonal antibodies directed against cell surface determinants on lymphoid cells of the mouse. This method employs Terasaki-type trays coated with a monolayer of target cells fixed with 0.02% glutaraldehyde. Cells are incubated with monoclonal antibodies, followed by affinity column-purified rabbit-anti-rat immunoglobulin antibodies and a protein-A-β-galactosidase conjugate. Binding of antibodies to the cells is visualized by incubation with the substrate 4-methylumbelliferyl galactoside. Fluorescence in the individual wells of the Terasaki trays is then quantitatively analysed within 120 s using a scanning inverted microfluorometer, connected to a digital voltmeter and a desk-top calculator.Keywords
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