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Clinical Chemistry 24: 1590-1594, 1978;
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Clinical Chemistry, Vol 24, 1590-1594, Copyright © 1978 by American Association for Clinical Chemistry

Double-antibody enzyme immunoassay for nortriptyline

MN Al-Bassam, MJ O'Sullivan, E Gnemmi, JW Bridges and V Marks

beta-D-Galactosidase (EC 3.2.1.23) from Escherichia coli was conjugated to desmethylnortriptyline by means of a bifunctional cross-linking reagent, dimethyl adipimidate, and used in a double-antibody immunoassay for nortriptyline. Eighty percent of the enzyme activity was retained after conjugation; 75% of the enzyme was conjugated to desmethylnortriptyline. In the final immunoassay the enzyme activity of the bound fraction was determined with o-nitrophenyl-beta-D- galactopyranoside as substrate. The sensitivity, precision, and simplicity of the enzyme immunoassay compared favorable with that of a published radioimmunoassay method. Results for nortriptyline in plasma samples correlated well with those determined by either radioimmunoassay or gas-chromatography.




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Copyright © 1978 by the American Association for Clinical Chemistry.