Catalyzed colorimetric and fluorometric substrates for peroxidas

Details Catalyzed colorimetric and fluorometric substrates for peroxidas Catalyzed colorimetric and fluorometric substrates for peroxidas

1982-09-22

1985-06-04

Warden, Robert J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving oxidoreductase

435805, 436135, 436826, 436904, C12Q 128, G01N 3352, G01N 3354

Patent

active

045215111

ABSTRACT:
Improved catalyzed substrates for use in developing characteristic colors or fluorescence in the presence of peroxidase enzymes (e.g., horseradish peroxidase) are disclosed which include as a rate accelerator a substituted phenol such as a p-halogenated phenol. The complete system typically includes a peroxide type oxidizing agent (e.g., hydrogen peroxide), a chromogenic or flurogenic compound (e.g., ABTS), a buffer and the accelerator compound. Advantageously, the accelerator should provide at least about 50 percent rate enhancement for the substrate, as compared with an otherwise identical, accelerator-free substrate reacted under the same conditions; however, the most preferred accelerator, p-iodophenol, gives enhancements on the order to 1,000 percent. The invention is particularly useful in so-called ELISA determinations which involve an enzyme-linked moiety, and permit detection at very low concentration levels unobtainable with conventional colorimetric substrates.

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Lyttle et al., Canadian Journal of Biochemistry 1973, vol. 51, pp. 1066-1071.
Pommier et al., The Journal of Biological Chemistry, vol. 254, No. 8, 1979, pp. 3006-3010.
Yeh et al., Canadian Journal of Biochemistry, vol. 49, 1971, pp. 163-165.

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