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Clinical Chemistry 39: 965-971, 1993;
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Clinical Chemistry, Vol 39, 965-971, Copyright © 1993 by American Association for Clinical Chemistry

Determination of one thousandth of an attomole (1 zeptomole) of alkaline phosphatase: application in an immunoassay of proinsulin

DB Cook and CH Self
Department of Clinical Biochemistry, Medical School, University of Newcastle upon Tyne, UK.

Enzyme amplification has proved to be a highly sensitive quantification technique for immunoassays. We have shown that by using a fluorescent end-point, even more sensitive enzyme amplification assays can be generated than hitherto reported. We describe some general properties of this system and demonstrate its application in an assay for human proinsulin in plasma. The detection system can be used to measure less than one thousandth of an attomole (1 zeptomole) of alkaline phosphatase, equivalent to about 350 molecules of alkaline phosphatase per well of a microtiter plate. We have used this system to construct a proinsulin assay with a sensitivity of 0.017 pmol/L.




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Copyright © 1993 by the American Association for Clinical Chemistry.