Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving hydrolase
Patent
1996-05-28
1998-12-08
Gitomer, Ralph
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving hydrolase
435 19, 422 56, C12Q 137
Patent
active
058467548
ABSTRACT:
The present invention involves a composition and method for the determination of enzymatic activity or enzyme substrate concentration in a fluid test sample in which the enzyme catalyzes a reaction in a reagent system to provide a detectable response indicative of the concentration of the enzyme or enzyme substrate in the test sample. A measured amount of a protease capable of eliminating the enzymatic activity is included in the reagent composition, so that when a measured amount of test sample is treated, the reaction producing the detectable response will have a definite end point. The invention is particularly useful in conjunction with the determination of leucocytes in urine wherein the leukocytes are lysed to release their human leucocyte elastase which hydrolyzes a chromogenic ester to provide a phenol which then reacts with a diazonium salt to form a colored azo dye.
REFERENCES:
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patent: 5464739 (1995-11-01), Johnson et al.
Cook et al., "Effect of Cu 2+ and Zn 2+ on the Inhibition of Human Leucocyte Elastase by 6-Alkyl-3-(.omega.-carboxy alkyl)-2-pyrones, Oleic Acid and Sulindac Sulfide," Biol. Chem. Hoppe Seyler 1989, 370, 11-19, Jan. 1989.
Pyun J., Effective Termination Method for Proteolytic Reaction Using Trypsin Immobilized on a PEI Cellulose TLC Strip, Biotechniques 19(5) 728-30, Nov. 1995.
Maunoury R., Mise en Evidence de la Centrosphere de Cellules Humaines par la Technique D'Immunoperoxydase Avec des Anticorps Naturels, Biol Cellulaire 36, 91-94, 1979.
Cook et al, Biol. Chem., vol. 370, pp. 11-19, (1989).
Pugia Michael J.
Schaeper Robert J.
Bayer Corporation
Gitomer Ralph
Jeffers Jerome L.
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