Enzymic method of detecting analytes and novel substrates theref

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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435 7, 435 18, 435 19, 435810, 436501, G01N 33535, G12Q 168

Patent

active

047450541

ABSTRACT:
A small enzymically inactive peptide fragment of an enzyme (e.g. ribonuclease S-peptide) is used as the label and conjugated with the complementary fragment (S-protein) to form an enzyme which catalyzes a primary reaction whose product is, or leads to, an essential coenzyme or prosthetic group for a second enzyme which catalyzes a secondary reaction leading to a detectable result indicating the presence of analyte. Also disclosed are novel synthetic substrates for the primary reaction. Substrates for ribonuclease S conjugate enzyme are of the formula R-X where R is a pyrimidine 3'-phosphate moiety and X is a leaving group linked to R through the 3'-phosphate group and leads to said coenzyme or prosthetic group, e.g. via riboflavin, thiamine, pyridoxal, pyridoxine or pyridoxine phosphate.

REFERENCES:
patent: 3852157 (1974-12-01), Rubenstein et al.
patent: 4378428 (1983-03-01), Farina et al.
patent: 4463090 (1984-07-01), Harris
patent: 4598042 (1986-07-01), Self

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