Dual enzyme labels for simultaneous heterogeneous enzyme-linked competitive binding assays

Abstract

The use of two enzyme labels in a dual analyte simultaneous heterogeneous enzyme-linked competitive binding assay is examined. Reaction conditions for monitoring glucosed-phosphate dehydrogenase (G6PDH) and Pgalactosidase (@gal) activities are found which enable the independent measurement of each enzyme activity in the presence of the other. The two enzymes are used as labels in the development of a model simultaneous heterogeneous competitive binding assay for detecting the biotin and vitamin B,, content of vitamin tablets. The simultaneous assay is shown to exhibit analytical dose-response characteristics comparable to those of the single analyte assays. The potential use of the G6PDHIpgal enzyme pair for devising other dual analyte competitive and non-competitive binding assays is discussed.

Simultaneous Determination of 13 Fluoroquinolone and 22 Sulfonamide Residues in Milk by a Dual-Colorimetric Enzyme-Linked Immunosorbent Assay

“…A multianalyte immunoassay that can measure two or more chemical classes in one well has become a long established goal. 27 The radioimmunoassay using I 131 and I 125 as labels was reported for the detection of human insulin and growth hormone in serum samples in 1968. 28 More recently, dual-label time-resolved fluorescence immunoassays 29 have been described for the simultaneous detection of two or more analytes.…”

“…These improvements make the multianalyte immunoassay the method of choice for screening samples for multianalytes. A multianalyte immunoassay that can measure two or more chemical classes in one well has become a long established goal …”