AU2001293768A1 - Process for detecting enzyme activity in an immunoassay - Google Patents

Process for detecting enzyme activity in an immunoassay

Info

Publication number
AU2001293768A1
AU2001293768A1 AU2001293768A AU9376801A AU2001293768A1 AU 2001293768 A1 AU2001293768 A1 AU 2001293768A1 AU 2001293768 A AU2001293768 A AU 2001293768A AU 9376801 A AU9376801 A AU 9376801A AU 2001293768 A1 AU2001293768 A1 AU 2001293768A1
Authority
AU
Australia
Prior art keywords
modified
enzyme activity
immunoassay
peptide
antibody
Prior art date
Legal status (The legal status 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 status listed.)
Abandoned
Application number
AU2001293768A
Inventor
Dr. Karsten Henco
Joachim Kramer
Thomas Mander
Christine Peiker
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Evotec OAI AG
Original Assignee
Evotec Biosystems GmbH
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
Priority claimed from EP00117457A external-priority patent/EP1182263A1/en
Priority claimed from EP00122707A external-priority patent/EP1199370B1/en
Priority claimed from EP00128176A external-priority patent/EP1217078A1/en
Application filed by Evotec Biosystems GmbH filed Critical Evotec Biosystems GmbH
Publication of AU2001293768A1 publication Critical patent/AU2001293768A1/en
Abandoned legal-status Critical Current

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
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
    • G01N33/6842—Proteomic analysis of subsets of protein mixtures with reduced complexity, e.g. membrane proteins, phosphoproteins, organelle proteins
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0804—Tripeptides with the first amino acid being neutral and aliphatic
    • C07K5/081—Tripeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/08—Tripeptides
    • C07K5/0802—Tripeptides with the first amino acid being neutral
    • C07K5/0812—Tripeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1005—Tetrapeptides with the first amino acid being neutral and aliphatic
    • C07K5/1013—Tetrapeptides with the first amino acid being neutral and aliphatic the side chain containing O or S as heteroatoms, e.g. Cys, Ser
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
    • C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
    • C07K5/10—Tetrapeptides
    • C07K5/1002—Tetrapeptides with the first amino acid being neutral
    • C07K5/1016—Tetrapeptides with the first amino acid being neutral and aromatic or cycloaliphatic
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/10—Transferases (2.)
    • C12N9/12—Transferases (2.) transferring phosphorus containing groups, e.g. kinases (2.7)
    • C12N9/1205—Phosphotransferases with an alcohol group as acceptor (2.7.1), e.g. protein kinases
    • 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
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/91—Transferases (2.)
    • G01N2333/91045—Acyltransferases (2.3)
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
    • G01N2333/90—Enzymes; Proenzymes
    • G01N2333/914—Hydrolases (3)
    • G01N2333/916—Hydrolases (3) acting on ester bonds (3.1), e.g. phosphatases (3.1.3), phospholipases C or phospholipases D (3.1.4)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Medicinal Chemistry (AREA)
  • Genetics & Genomics (AREA)
  • Biophysics (AREA)
  • Biomedical Technology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Immunology (AREA)
  • Urology & Nephrology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Hematology (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Pathology (AREA)
  • Bioinformatics & Computational Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Analytical Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • General Engineering & Computer Science (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

Detection of hydrolytic enzyme activity in an immunoassay comprises incubating a dually modified luminescent labelled protein/peptide, comprising a first modified position as a substrate for an enzyme and a second modified position as an affinity enhancer for an antibody, and adding an antibody discriminating the dually modified substrate from the enzyme generated product. The method is useful for detecting enzyme activity in an immunoassay of phosphatases PP2A and PP1 using TAMRA-Iabelled bisphosphorylated peptides, and of Histone H3 acetylase or deacetylase activity using TAMRA-labelled bisacetylated-monophosphorylated peptides. In addition the method is useful for screening modulators for enzyme activity. For the histone H3 (de)acetylase assay a bisacetylated labelled peptide can only be sufficiently discriminated by an antibody from the monoacetylated substrate if a third modified residue i.e. a phosphorylated residue is present in the peptide as affinity enhancer.
AU2001293768A 2000-08-11 2001-08-13 Process for detecting enzyme activity in an immunoassay Abandoned AU2001293768A1 (en)

Applications Claiming Priority (9)

Application Number Priority Date Filing Date Title
EP00117457A EP1182263A1 (en) 2000-08-11 2000-08-11 Process for detecting threonine/serine kinase activity
EP00117457.2 2000-08-11
EP00118431 2000-08-24
EP00118431.6 2000-08-24
EP00122707A EP1199370B1 (en) 2000-10-18 2000-10-18 Process for detecting dephosphorylation of phosphorylated threonine or serine by phosphatase activity
EP00122707.3 2000-10-18
EP00128176A EP1217078A1 (en) 2000-12-21 2000-12-21 Phosphatase immunoassay using antibodies against bisphosphorylated substrates
EP00128176.5 2000-12-21
PCT/EP2001/009354 WO2002014543A2 (en) 2000-08-11 2001-08-13 Process for detecting enzyme activity in an immunoassay

Publications (1)

Publication Number Publication Date
AU2001293768A1 true AU2001293768A1 (en) 2002-02-25

Family

ID=27439967

Family Applications (1)

Application Number Title Priority Date Filing Date
AU2001293768A Abandoned AU2001293768A1 (en) 2000-08-11 2001-08-13 Process for detecting enzyme activity in an immunoassay

Country Status (5)

Country Link
EP (1) EP1313876B8 (en)
AT (1) ATE350485T1 (en)
AU (1) AU2001293768A1 (en)
DE (1) DE60125783T2 (en)
WO (1) WO2002014543A2 (en)

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1670943B1 (en) 2003-09-30 2013-11-13 Evotec International GmbH Diagnostic and therapeutic use of a sulfotransferase for alzheimer's disease
US20080293081A1 (en) * 2005-03-03 2008-11-27 Sirtris Pharmaceuticals, Inc. Fluorescence Polarization Assays for Acetyltransferase/Deacetylase Activity
WO2006128902A1 (en) 2005-06-01 2006-12-07 Evotec Neurosciences Gmbh Diagnostic and therapeutic target slc39a12 proteins for neurodegenerative diseases
EP1891241B1 (en) 2005-06-16 2011-01-12 EVOTEC Neurosciences GmbH Diagnostic and therapeutic target adarb2 proteins for neurodegenerative diseases
WO2007060599A1 (en) * 2005-11-25 2007-05-31 Koninklijke Philips Electronics N.V. Magnetic biosensor for determination of enzymic activity
WO2007064902A2 (en) * 2005-12-02 2007-06-07 Sirtris Pharmaceuticals, Inc. Mass spectrometry assays for acetyltransferase/deacetylase activity
EP2053397A1 (en) 2007-10-25 2009-04-29 EVOTEC Neurosciences GmbH PPM1E proteins and nucleic acids as targets for neurodegenerative diseases

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ATE233325T1 (en) * 1997-12-05 2003-03-15 Upjohn Co FLUORESCENCE-BASED HTS (HIGH THROUGHPUT SCREENING) ASSAY METHODS FOR PROTEIN KINASE AND PHOSPHATASE

Also Published As

Publication number Publication date
WO2002014543A2 (en) 2002-02-21
DE60125783D1 (en) 2007-02-15
DE60125783T2 (en) 2007-10-18
EP1313876B8 (en) 2007-02-28
EP1313876A2 (en) 2003-05-28
ATE350485T1 (en) 2007-01-15
WO2002014543A3 (en) 2003-03-20
EP1313876B1 (en) 2007-01-03

Similar Documents

Publication Publication Date Title
Knisatschek et al. Characterization of “thyroliberin-deamidating enzyme” as a post-proline-cleaving enzyme. Partial purification and enzyme-chemical analysis of the enzyme from anterior pituitary tissue.
Yang et al. Rubiscolin, a δ selective opioid peptide derived from plant Rubisco
JP3727269B2 (en) Method for identifying methionine aminopeptidase inhibitors
AU6715090A (en) Enzyme amplified lanthanid chelate luminescence assay
WO2003036256A3 (en) Method and apparatus for increasing the dynamic range and accuracy of binding assays
JP2001514506A (en) Protease inhibitor analysis
AU3625999A (en) Detection of a target in a sample
CA2451909A1 (en) High throughput assays for the proteolytic activities of clostridial neurotoxins
Ng et al. A fluorescent oligopeptide energy transfer assay with broad applications for neutral proteases
WO2005042771A3 (en) Colorimetric substrates, colorimetric sensors, and methods of use
WO2003021226A3 (en) Peptide biomarker and method of identification
AU2001293768A1 (en) Process for detecting enzyme activity in an immunoassay
Korecká et al. Utilization of newly developed immobilized enzyme reactors for preparation and study of immunoglobulin G fragments
AU2126400A (en) Detection of small molecules by use of a piezoelectric sensor
Dick et al. Identification and localization of a cysteinyl residue critical for the trypsin-like catalytic activity of the proteasome
WO2003012393A3 (en) Method of detecting protease activity in a cell
WO2003008635A3 (en) η-SECRETASE IN VITRO SCREENING ASSAY
DE60026598D1 (en) NEW ENZYMATIC SUBSTRATES FOR THE DETECTION OF PSEUDOMONAS AERUGINOSA
Barros et al. Neprilysin carboxydipeptidase specificity studies and improvement in its detection with fluorescence energy transfer peptides
Martin et al. Carpe diubiquitin
Norman et al. Proteolytic conversion of [Met] enkephalin-Arg6-Gly7-Leu8 by brain synaptic membranes. Characterization of formed peptides and mechanism of proteolysis.
WO2002014542A3 (en) Process for detecting serine/threonine kinase activity
Stebbins et al. A microtiter colorimetric assay for the HIV-1 protease
Consalvo et al. Rapid fluorimetric assay for the detection of the peptidyl α-amdating enzyme intermediate using high-performance liquid chromatography
WO2001038560A3 (en) Active enzyme detection using immobilized enzyme inhibitors