Method of determining enzymatic activity in biological media

Details Method of determining enzymatic activity in biological media Method of determining enzymatic activity in biological media

C - Chemistry, Metallurgy

12

Q

C12Q 1/56 (2006.01)

Patent

CA 2654474

A reliable, fast and convenient method is provided to determine the change in concentration of enzyme in time in a biological medium in the presence of a signal substrate that corrects for substrate consumption, color-dependency of the signal and non-linearity between the concentration of the leaving group of the signal substrate and the amount of signal. The method comprises the measurement of a calibrator curve in a suitable medium, such as a buffer, to determine the characteristics of the measured curve. These characteristics will then allow to obtain this whole curve or a sufficient part thereof again through a mathematical procedure based on a single-point measurement in a sample of a medium in which the enzyme generation takes place. This has the great advantage that there is no need to measure the whole calibrator curve in each individual medium, since a single-point measurement is adequate.

Cette invention concerne un procédé rapide et pratique permettant de déterminer le changement de concentration en fonction du temps d'une enzyme dans un milieu biologique, en présence d'un substrat de signal permettant de corriger la consommation due au substrat, la dépendance du signal à la couleur et la non-linéarité entre la concentration du groupe labile du substrat de signal et la valeur du signal. Ledit procédé implique de mesurer une courbe d'étalonnage dans un milieu approprié, par exemple un tampon, afin de déterminer les caractéristiques de la courbe mesurée. Ces caractéristiques permettent ensuite d'obtenir de nouveau ladite courbe entière, ou une portion suffisante de ladite courbe, au moyen d'une procédure mathématique basée sur le mesurage en un point d'un échantillon d'un milieu dans lequel a lieu une génération d'enzyme. Un tel procédé présente le grand avantage de ne pas avoir à mesurer l'intégralité de la courbe d'étalonnage dans chacun des milieux, étant donné qu'une mesure en un point suffit.

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