Protein isolation and purification

Details Protein isolation and purification Protein isolation and purification

C - Chemistry, Metallurgy

12

N

C12N 15/82 (2006.01) A23J 1/14 (2006.01) C07K 1/14 (2006.01) C12N 15/62 (2006.01)

Patent

CA 2589158

A method for purifying a recombinant fusion protein that is expressed as recombinant protein body-like assemblies (RPBLAs) in host cells is disclosed in which an aqueous homogenate of transformed host cells that express a fusion protein as RPBLAs having a predetermined density is provided. Regions of different density are formed in the homogenate to provide a region that contains a relatively enhanced concentration of the RPBLAs and a region that contains a relatively depleted concentration of the RPBLAs. The RPBLAs- depleted region is separated from the region of relatively enhanced concentration of RPBLAs, thereby purifying said fusion protein. The region of relatively enhanced concentration of RPBLAs can thereafter be collected as desired.

L'invention concerne un procédé permettant de purifier une protéine de fusion recombinante exprimée sous forme d'ensembles du type corps de protéine recombinante (RPBLA) dans des cellules hôtes dans lesquelles un broyat aqueux de cellules hôtes transformées exprime une protéine de fusion telle que RPBLA possédant une densité prédéterminée. Des régions de densités différentes sont formées dans le broyat afin de fournir une région qui contient une concentration de RPBLA relativement améliorée et une région qui contient une concentration de RPBLA relativement appauvrie. La région appauvrie en RPBLA est séparée de la région à concentration de RPBLA relativement améliorée, ce qui permet de purifier la protéine de fusion. La région à concentration de RPBLA relativement améliorée peut ensuite être collectée le cas échéant.

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