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Clinical Chemistry 31: 1546-1549, 1985;
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Clinical Chemistry, Vol 31, 1546-1549, Copyright © 1985 by American Association for Clinical Chemistry

Heterogeneous enzyme immunoassay with electrochemical detection: competitive and "sandwich"-type immunoassays

KR Wehmeyer, HB Halsall and WR Heineman

In these competitive and "sandwich"-type heterogeneous enzyme immunoassays, based on liquid chromatography with electrochemical detection, rabbit immunoglobulin G is used as a model compound. Alkaline phosphatase (EC 3.1.3.1), the labeling enzyme, catalyzes conversion of phenyl phosphate to phenol. After separation on an octyldecylsilane column, the enzyme-generated phenol is detected in a thin-layer cell at a carbon-paste working electrode. The detection limit for phenol is 5.0 nmol/L. The electrode response varies linearly with concentration over a range of three orders of magnitude. For the sandwich-type assay procedure the detection limit is 10 ng/L; the linearity ranges over four orders of magnitude. The detection limit of the competitive immunoassay is 5 micrograms/L. The dynamic range spans two orders of magnitude.




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Copyright © 1985 by the American Association for Clinical Chemistry.