Method for measuring the concentration of protease inhibitors, k

Details Method for measuring the concentration of protease inhibitors, k Method for measuring the concentration of protease inhibitors, k

1997-06-20

1999-01-05

Leary, Louise N.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving hydrolase

435 24, 435 4, 435963, 435183, 435975, C12Q 137, C12Q 100, G01N 3353

Patent

active

058561170

ABSTRACT:
This invention provides a method for measuring the concentration of urinary trypsin inhibitors which is excellent in precision and reproducibility, whose operation is simple, and in which a possibility of damaging a plastic cell is eliminated. The method for measuring the concentration of urinary trypsin inhibitors comprises mixing an urine sample, a protease solution containing trypsin, and a buffer solution, adding a substrate solution to the mixture to cause the enzyme reaction, and measuring the activity of the enzyme, wherein the buffer solution is prepared so that it contains at least 0.15 .mu.mol calcium per 1 .mu.g of the trypsin but no more than 100 .mu.mol calcium per 1 ml of the urine sample in the reaction mixture, and wherein the substrate solution is prepared by dissolving the substrate in an organic solvent and diluting the mixture solution with aqueous medium, wherein at least one of an amphoteric surfactant and a nonionic surfactant is added to at least one of the organic solvent and the aqueous medium.

REFERENCES:
patent: 5354491 (1994-10-01), Bjorkquist et al.
H.J. Faarvang, "Excretion of Mingin in Urine in Normal and in Pathological Conditions", Scandinav. J. Clin. & Lab. Investigation 17 (Suppl. 83), pp. 25-28, 1965.
"Measurement of Alpha.sub.1 -Antitrypsin in Serum, by Immunodiffusion and by Enzymatic Assay", Clinical Chemistry, vol. 20, No. 3, pp. 396-399, 1974.
Beatrice Kassell, "Bovine Trypsin-Kallikrein Inhibitor (Kunitz Inhibitor, Basic Pancreatic Trypsin Inhibitor, Polyvalent Inhibitor from Bovine Organs)", Methods of Enzymatic Analysis, 3rd Ed., vol. 10, pp. 844-845, 1970. Month not available.
Beatric Kassell, "Protoeolytic Enzyme Inhibitors from Ascaris lumbricoides", Methods of Enzymatic Analysis, 3rd Ed., vol. 10, pp. 872-873, 1970. Month not available.
Shiro Kuwajima, et al., "Urinary trypsin inhibitor and its clinical usefulness for diagnosis of acute phase reactant and renal disease", Japanese Journal of Inflammation Review Article, vol. 9, No. 3, pp. 175-182, May 1989.
Shiro Kuwajima, et al., "Automated Measurement of Trypsin Inhibitor in Urine with a Centrifugal Analyzer: Comparison with Other Acute Phase Reactants", Clinical Biochemistry, vol. 23, pp. 167-171, Apr. 1990.
T. Bogacheva et al., "Method for Determining the Molar Concentration of Protease Inhibitors and Their Enzyme Affinity", Chemical Abstracts, vol. 98, No. 5, p. 294 (Jan. 31, 1983).
European Search Report (Sep. 23, 1997).

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