Method for isolating proteins or protein and nucleic acid...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/10 (2006.01) B03C 1/01 (2006.01) C07K 1/22 (2006.01) G01N 33/543 (2006.01)

Patent

CA 2412536

The invention concerns a method for isolating proteins and/or protein and nucleic acid associations in a sample, comprising steps which consist in: contacting said sample and magnetic colloidal particles comprising a core and a coat wherein: the core is magnetic and is coated with at least a polymer comprising functional groups X selected among amine, hydroxyl, thiol, aldehyde, ester, anhydride, acid chloride, carbonate, carbamate, isocyanate and isothiocyanate groups or mixtures thereof, whereof at least one fraction has reacted with other functional groups of the coat, and the coat consists of a polymer bearing functional groups Z and Z', capable of ionisation, identical or different, selected among amine, carboxylic acid, ester, anhydride, aldehyde, thiol, disulphide, .alpha.-halogenocarbonyl, sulphonic acid, maleimide, isocyanate and isothiocyanate groups to form a mixture; incubating said mixture in predetermined conditions; and separating from the mixture the protein and/or protein and nucleic acid associations complexed on the colloidal particles by applying a magnetic field. The invention also concerns a complex consisting of colloidal particles and proteins, a reagent comprising such a complex or colloidal particles.

Le procédé d'isolement de protéines et/ou d'une association de protéines et d'acides nucléiques dans un échantillon comprend les étapes selon lesquelles: on met en contact ledit échantillon et des particules colloïdales magnétiques comprenant un coeur et une enveloppe dans lesquelles: le coeur est magnétique et est enrobé par au moins un polymère comprenant des groupements fonctionnels X choisis parmi les groupements amine, hydroxyle, thiol, aldéhyde, ester, anhydride, chlorure d'acide , carbonate, carbamate, isocyanate et isothiocyanate ou leurs mélanges, dont au moins une fraction a réagi avec d'autres groupements fonctionnels de l'enveloppe, et l'enveloppe est constituée d'un polymère portant des groupements fonctionnels ionisables, Z et Z', identiques ou différents, choisis parmi les groupements amine, acide carboxylique, ester, anhydride, aldéhyde, thiol, disulfure, .alpha.- halogénocarbonyle, acide sulfonique, maléimide, isocyanate, et isothiocyanate, qui ont partiellement réagi avec les groupements fonctionnels X du coeur; pour constituer un mélange, on incube ledit mélange sous des conditions prédéterminées, et on sépare du mélange les protéines et/ou les associations de protéines et d'acides nucléiques complexées sur lesdites particules colloïdales par application d'un champ magnétique. L'invention concerne aussi un complexe formé de particules colloïdales et de protéines, un réactif comprenant un tel complexe ou des particules colloïdales.

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