Methods and compositions for rapid protein and peptide...

C - Chemistry – Metallurgy – 07 – K

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C07K 1/34 (2006.01) B01J 20/32 (2006.01) C12M 1/33 (2006.01) C12N 1/06 (2006.01) C12N 9/22 (2006.01) C12P 21/00 (2006.01) G01N 30/60 (2006.01)

Patent

CA 2415713

The present invention relates generally to compositions, methods and kits for use in extracting and isolating protein or peptide molecules. More specifically, the invention relates to such compositions, methods and kits that are useful in the isolation of protein or peptide molecules from cells (e.g., bacterial cells, animal cells, fungal cells, viruses, yeast cells or plant cells) via lysis and one or more additional isolation procedures, such as one or more filtration and/or chromatography procedures. In particular, the invention relates to compositions, methods and kits wherein protein or peptide molecules are isolated using an integrated lysis/filtration matrix, which may comprise one or more supports (e.g., polyolefin, scintered polyethylene, nitrocellulose, polypropylene, polycarbonate, cellulose acetate, silica, and the like). The compositions, methods and kits of the invention are suitable for isolating a variety of forms of protein or peptide molecules from cells. The compositions, methods and kits of the invention are particularly well- suited for rapid isolation of recombinant protein or peptide molecules expressed in bacterial cells, either as soluble protein, or as an inclusion body. The invention is particularly useful in high throughput applications, allowing quick isolation and/or analysis of proteins and/or peptides from numerous sources.

La présente invention concerne généralement des compositions, des méthodes et des kits utilisés pour extraire et isoler des molécules de peptides ou de protéines. Plus spécifiquement, cette invention concerne ces compositions, méthodes et kits que l'on utilise dans l'isolation de molécules de peptides ou de protéines à partir de cellules (par exemple, des cellules bactériennes, animales, fongiques, des virus, des cellules de levure ou des cellules végétales) par lyse et au moins une procédure d'isolation complémentaire, telle que des procédures de filtration et/ou de chromatographie. Notamment, cette invention concerne des compositions, des méthodes et des kits au moyen desquels des molécules de peptides ou de protéines sont isolées à l'aide d'une matrice de filtration/de lyse intégrée qui peut renfermer au moins un support (par exemple, la polyoléfine, le polyéthylène fritté, le nitrocellulose, le polypropylène, le polycarbonate, l'acétate de cellulose, la silice et similaire). Les compositions, méthodes et kits de cette invention sont appropriés à l'isolation d'une variété de formes de molécules de peptides ou de protéines à partir de cellules. Ces compositions, méthodes et kits conviennent spécifiquement à l'isolation rapide de molécules de peptides ou de protéines recombinantes exprimées dans des cellules bactériennes comme protéines solubles ou corps d'inclusion, ainsi que dans des applications à rendement élevé, ce qui permet une isolation et/ou une analyse rapides des protéines et/ou des peptides à partir de plusieurs sources.

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