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1. (WO1992010761) METHOD AND REAGENTS FOR THE DETERMINATION OF THE ENZYMIC ACTIVITY OF TRANSGLUTAMINASE (EC 2.3.2.13)
Latest bibliographic data on file with the International Bureau

Pub. No.: WO/1992/010761 International Application No.: PCT/EP1991/002223
Publication Date: 25.06.1992 International Filing Date: 26.11.1991
Chapter 2 Demand Filed: 25.06.1992
IPC:
C12Q 1/48 (2006.01) ,C12Q 1/52 (2006.01) ,C12Q 1/56 (2006.01) ,G01N 33/573 (2006.01) ,G01N 33/86 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
48
involving transferase
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
48
involving transferase
52
involving transaminase
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
Q
MEASURING OR TESTING PROCESSES INVOLVING ENZYMES OR MICRO-ORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
1
Measuring or testing processes involving enzymes or micro-organisms; Compositions therefor; Processes of preparing such compositions
56
involving blood clotting factors, e.g. involving thrombin, thromboplastin, fibrinogen
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
50
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
53
Immunoassay; Biospecific binding assay; Materials therefor
573
for enzymes or isoenzymes
G PHYSICS
01
MEASURING; TESTING
N
INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
33
Investigating or analysing materials by specific methods not covered by groups G01N1/-G01N31/131
48
Biological material, e.g. blood, urine; Haemocytometers
50
Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
86
involving blood coagulating time
Applicants:
DEMPFLE, Carl-Erik [DE/DE]; DE
Inventors:
DEMPFLE, Carl-Erik; DE
Agent:
ZELLENTIN UND PARTNER; Rubensstraße 30 D-6700 Ludwigshafen, DE
Priority Data:
P 40 38 899.906.12.1990DE
Title (DE) VERFAHREN UND REAGENZIEN ZUR BESTIMMUNG DER ENZYMATISCHEN AKTIVITÄT VON TRANSGLUTAMINASEN (EC 2.3.2.13)
(EN) METHOD AND REAGENTS FOR THE DETERMINATION OF THE ENZYMIC ACTIVITY OF TRANSGLUTAMINASE (EC 2.3.2.13)
(FR) PROCEDE ET REACTIFS POUR LA DETERMINATION DE L'ACTIVITE ENZYMATIQUE DES TRANSGLUTAMINASES (EC 2.3.2.13)
Abstract:
(DE) Die vorliegende Erfindung betrifft ein Verfahren zur Bestimmung des Blutgerinnungsfaktors XIII (Plasma-Transglutaminase und Thrombocyten-Transglutaminase) und anderer Transglutaminasen der Enzymgruppe EC 2.3.2.13, wobei a) Microtiterplatten aus Kunststoffen oberflächlich mit Proteinen oder Peptiden beschichtet werden, welche Glutamin enthalten; b) Calciumchlorid, eine als Reduktionsmittel wirkende Thioverbindung, eine Verbindung mit einer primären Aminogruppe der allgemeinen Formel (I): NH2-(CH2)n-X, in welcher n eine ganze Zahl von 4-8 und X eine inerte, mit spezifischen Antikörpern koppelnde Gruppe bedeutet, sowie eine Transglutaminase-haltige Probe auf der beschichteten Microtiterplatte zur Reaktion gebracht werden, wobei gegebenenfalls Thrombin zur proteolytischen Aktivierung der Transglutaminase zugefügt wird; c) die durch Transglutaminase an die Platte gebundenen Amine mit einem markierten Antikörper gegen die Gruppe X umgesetzt; d) die Menge des gebundenen Amins bzw. des Antikörpers mittels eines geeigneten Nachweisreagenzes bestimmt; e) und jeweils zwischen den einzelnen Schritten gelöste Reagenzreste ausgewaschen werden sowie ein Reagenzkit zur Durchführung des Verfahrens.
(EN) The invention concerns a method for the determination of blood-clotting factor XIII (plasma transglutaminase and thrombocyte transglutaminase) and other transglutaminases of the EC 2.3.2.13 group of enzymes. The method comprises the following steps: a) plastic microtitre plates are surface-coated with proteins or peptides containing glutamine; b) calcium chloride, a thio-compound acting as a reducing agent, a compound, containing a primary amino group, of general formula NH2-(CH2)n-X in which n is a whole number from 4 to 8 and X is an inert group which couples with specific antibodies, plus a probe containing transglutaminase are reacted together on the coated microtitre plate, thrombin optionally being added for the proteolytic activation of the transglutaminase; c) the amines bound by the transglutaminase to the plate are reacted with a marked antibody active against the group X; d) the amount of bound amine or antibody is determined by means of a suitable analytical reagent; e) between each of these individual steps, dissolved reagent residues are washed out. The invention also concerns a reagent kit for carrying out the method.
(FR) L'invention concerne un procédé pour la détermination du facteur de coagulation XIII (plasma-transglutaminase et thrombocytes-transglutaminase) et d'autres transglutaminases du groupe enzymatique EC 2.3.2.13, procédé selon lequel: a) des plaques de microtitrage en matière plastique sont garnies, en surface, de protéines ou de peptides renfermant de la glutamine; b) du chlorure de calcium, un composé thio agissant comme réducteur, un composé présentant un groupe amino primaire de formule NH2-(CH2)n-X dans laquelle n est un nombre entier compris entre 4 et 8, et X désigne un groupe inerte susceptible de s'associer à des anticorps spécifiques, ainsi qu'une prise d'essai contenant de la transglutaminase, sont amenés à réagir sur la plaque garnie de microtitrage, réaction au cours de laquelle de la thrombine est éventuellement ajoutée en vue de l'activation protéolytique de de la transglutaminase; c) l'amine liée à la plaque, sous l'action de la transglutaminase, est mise à réagir sur le groupe X avec un anticorps marqué; d) la quantité d'amine liée, ou éventuellement d'anticorps, est déterminée à l'aide d'un réactif indicateur approprié; e) les restes de réactifs dissous étant éliminés par lavage entre chacune des différentes étapes. L'invention concerne également un jeu de réactifs pour la mise en ÷uvre de ce procédé.
Designated States: CA, US
European Patent Office (AT, BE, CH, DE, DK, ES, FR, GB, GR, IT, LU, NL, SE)
Publication Language: German (DE)
Filing Language: German (DE)