WO2001038560A3 - Active enzyme detection using immobilized enzyme inhibitors - Google Patents

Active enzyme detection using immobilized enzyme inhibitors Download PDF

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Publication number
WO2001038560A3
WO2001038560A3 PCT/US2000/032315 US0032315W WO0138560A3 WO 2001038560 A3 WO2001038560 A3 WO 2001038560A3 US 0032315 W US0032315 W US 0032315W WO 0138560 A3 WO0138560 A3 WO 0138560A3
Authority
WO
WIPO (PCT)
Prior art keywords
enzyme
inhibitor
mutant
active form
inhibitors
Prior art date
Application number
PCT/US2000/032315
Other languages
French (fr)
Other versions
WO2001038560A2 (en
Inventor
Daniel A Lawrence
Duane Day
Original Assignee
American Nat Red Cross
Daniel A Lawrence
Duane Day
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by American Nat Red Cross, Daniel A Lawrence, Duane Day filed Critical American Nat Red Cross
Priority to AU18011/01A priority Critical patent/AU1801101A/en
Publication of WO2001038560A2 publication Critical patent/WO2001038560A2/en
Publication of WO2001038560A3 publication Critical patent/WO2001038560A3/en

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/573—Immunoassay; Biospecific binding assay; Materials therefor for enzymes or isoenzymes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/56—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen

Abstract

The present method utilizes the capture of the functionally active form of an enzyme that covalently binds or binds with a dissociation constant of 1 x 10-9M or less to an enzyme inhibitor or mutant thereof. The present invention is directed to a method for detecting a functionally active form of an enzyme in a biological sample, comprising contacting an enzyme inhibitor or mutant thereof immobilized on a solid substrate with the biological sample, and measuring the binding of the enzyme inhibitor or mutant thereof to the active form of the enzyme by a detectable label. Specific enzyme inhibitors or mutants thereof are designed to covalently bind to specific clinically important enzymes. These enzyme inhibitors contain modifications that affect the binding to a target enzyme or affect the stability or the inhibitor. The nethod is particularly useful in measuring the presence of enzymes, such as tPA, elastase, cathepsin G and prostate specific antigen.
PCT/US2000/032315 1999-11-22 2000-11-22 Active enzyme detection using immobilized enzyme inhibitors WO2001038560A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU18011/01A AU1801101A (en) 1999-11-22 2000-11-22 Novel detection method for a functionally active form of an enzyme in biologicalsamples and a kit

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US16755399P 1999-11-22 1999-11-22
US60/167,553 1999-11-22

Publications (2)

Publication Number Publication Date
WO2001038560A2 WO2001038560A2 (en) 2001-05-31
WO2001038560A3 true WO2001038560A3 (en) 2002-01-10

Family

ID=22607840

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2000/032315 WO2001038560A2 (en) 1999-11-22 2000-11-22 Active enzyme detection using immobilized enzyme inhibitors

Country Status (2)

Country Link
AU (1) AU1801101A (en)
WO (1) WO2001038560A2 (en)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB0111987D0 (en) * 2001-05-16 2001-07-04 Univ Sheffield Method
WO2010006605A2 (en) * 2008-07-16 2010-01-21 Radiometer Medical Aps Thrombin substrate and assay for determining the level of bioactive thrombin in a sample
WO2012166755A1 (en) * 2011-06-02 2012-12-06 Lonza Walkersville, Inc. Enzyme-linked immunosorbent assay for quantitation of residual trypsin in a biological sample
GB2504499A (en) * 2012-07-31 2014-02-05 Baxter Healthcare Sa Selective measurement of active human protease coagulation factors
CN113092758B (en) * 2021-04-01 2022-11-08 山西集创生物科技有限公司 Double-antibody sandwich ELISA (enzyme-Linked immuno sorbent assay) detection kit for detecting trypsin analogue TrypLE (trypsin-like enzyme-linked immunosorbent assay), detection method and application
CN117169519B (en) * 2023-10-26 2024-01-30 艾康生物技术(杭州)有限公司 Dissociation agent and kit for detecting TT3 and/or TT4 in sample

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1990003577A1 (en) * 1988-09-30 1990-04-05 The University Of Vermont And State Agricultural College Immunoassays for catalytically-active, serine proteases
WO1990005309A1 (en) * 1988-11-04 1990-05-17 Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno A method of assaying an enzyme and kit and substances suitable for that method
US5110736A (en) * 1985-08-01 1992-05-05 Mitsui Toatsu Chemicals, Incorporated Purificaiton procedure of tpa from crude preparations
GB2259362A (en) * 1991-09-07 1993-03-10 Biosyn Limited Bio immunoassay for proteases

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5110736A (en) * 1985-08-01 1992-05-05 Mitsui Toatsu Chemicals, Incorporated Purificaiton procedure of tpa from crude preparations
WO1990003577A1 (en) * 1988-09-30 1990-04-05 The University Of Vermont And State Agricultural College Immunoassays for catalytically-active, serine proteases
WO1990005309A1 (en) * 1988-11-04 1990-05-17 Nederlandse Organisatie Voor Toegepast-Natuurwetenschappelijk Onderzoek Tno A method of assaying an enzyme and kit and substances suitable for that method
GB2259362A (en) * 1991-09-07 1993-03-10 Biosyn Limited Bio immunoassay for proteases

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
DATABASE BIOSIS [online] BIOSCIENCES INFORMATION SERVICE, PHILADELPHIA, PA, US; 1997, EHNEBOM J ET AL: "Comparison of chromogenic substrates for tissue plasminogen activator and the effects on the stability of plasminogen activator inhibitor type-1.", XP002174725, Database accession no. PREV199800119975 *
FIBRINOLYSIS & PROTEOLYSIS, vol. 11, no. 5-6, 1997, pages 287 - 293 *
ZHERDEV A V ET AL: "An enzyme immunoassay of catalytically active proteases.", ANALYTICAL LETTERS, vol. 25, no. 12, 1992, pages 2199 - 2208, XP001012095, ISSN: 0003-2719 *

Also Published As

Publication number Publication date
AU1801101A (en) 2001-06-04
WO2001038560A2 (en) 2001-05-31

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