Patent FR-2872290-A1 (FR)
Title: Implementation of plurality of parallel enzymatic reaction, used to diagnose e.g. viral infection, comprises obtaining solid support comprising zone matrix, depositing substrate and implementing plurality of the reaction in reactive volume
Inventor: BALAKIREV MAXIM
IPC: B01L3/00
Filed: 2004-06-24
Abstract: Process of implementation of a plurality of parallel enzymatic reactions in separated reactive volumes, comprises obtaining a solid support comprising a matrix of hydrophilic zones, where each hydrophilic zone is delimited by a hydrophobic zone; depositing or fixing a substrate in each hydrophilic zone; and implementing a plurality of parallel enzymatic reactions in reactive volume and comprising appropriate reactants and buffers. Process of implementation of a plurality of parallel enzymatic reactions in separated reactive volumes, comprises obtaining a solid support comprising a matrix of hydrophilic zones, where each hydrophilic zone is delimited by a hydrophobic zone (which is separated from other hydrophilic zones, hydrophilic and hydrophobic zones) comprising an aqueous solution deposited at the level of an hydrophilic zone forming a drop confined by the surface tension in the hydrophilic zone; depositing or fixing at least a substrate (which react in an enzymatic reaction) in each hydrophilic zone; and implementing a plurality of parallel enzymatic reactions in reactive volume (constituted by drops of aqueous solution confined in each hydrophilic zone of the support) and comprising appropriate reactants and buffers. Independent claims are also included for: (1) process of detection of parallel enzymatic activity in several samples, preferably several biological samples, comprising implementation of the process, in which the reactive mixture of each drop contains a sample (having an enzymatic activity) to be detected; (2) process of high throughput screening of inhibitor or regulators of an enzymatic activity, comprising the implementation of the process, in which the reactive mixture of each drop contains a reactive (having an enzymatic activity) in the presence of a compound, where the inhibiting or regulatory activity of the compound is researched; (3) process of preparation of a support for the implementation of the process, comprising depositing a mask containing glycerol on an hydrophilic solid support comprising glycerol/sucrose/water mixture in proportions such that the solution obtained is compatible with the piezoelectric exemption, in particular in terms of viscosity; and depositing a layer of hydrophobic silane by co-evaporation with pentane in inert atmosphere, under conditions allowing the formation of a layer of hydrophobic silane in the hydrophobic zone; optionally thermally stabilizing or dehydrating the layer of hydrophobic silane under vacuum; and (4) support comprising a matrix of hydrophilic zones, where each hydrophilic zone is delimited by a hydrophobic zone, which is separated from the other hydrophilic zones, the hydrophilic and hydrophobic zones (which is defined when an aqueous solution deposited on the level of an hydrophilic zone forms a drop confined by the surface tension in the hydrophilic zone), where the support comprises at least a substrate of an enzyme which is fixed to the surface in each of the hydrophilic zones.