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Inorganic pyrophosphatase as a label in heterogeneous enzyme immunoassay

✍ Scribed by A.A. Baykov; V.N. Kasho; S.M. Avaeva

Publisher
Elsevier Science
Year
1988
Tongue
English
Weight
475 KB
Volume
171
Category
Article
ISSN
0003-2697

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✦ Synopsis

Inorganic pyrophosphatase from Escherichia coli has been employed as a label in heterogeneous enzyme immunoassays. Enzyme-antibody conjugates were prepared with the use of glutaraldehyde and purified by gel permeation chromatography. Enzyme activity was measured by means of a sensitive one-step color reaction between phosphate, molybdate, and malachite green. The sensitivity in terms of absorbance readings was four to eight times higher than that of peroxidase-based assays. The color change (yellow to greenish blue) inherent in the use of pyrophosphatase as the labeling agent is highly suitable for visual analysis. Other merits of pyrophosphatase include the remarkable stability of the enzyme and its substrate, its compatibility with bacteriostatic agents, and its low Michaelis constant. Examples of the use of phosphatase in the assay of human alpha-fetoprotein and immunoglobulin G are presented.

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