Process for purifying primate gm-csf protein

Details Process for purifying primate gm-csf protein Process for purifying primate gm-csf protein

C - Chemistry, Metallurgy

07

K

530/13, 195/78,

C07K 14/535 (2006.01) C07K 1/16 (2006.01) C12N 15/70 (2006.01) C12N 15/81 (2006.01) C12N 15/85 (2006.01) A61K 38/00 (2006.01)

Patent

CA 1340852

A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises: preparing RNA from a cell that produces CSF; preparing polyadenylated messenger RNA from said RNA; preparing single stranded cDNA from said messenger RNA; converting the single stranded cDNA to double stranded cDNA; inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies; picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool; transfecting the plasmid DNA into suitable host cells for expressing CSF protein; culturing the transfected cells and assaying the supernatant for CSF activity; and selecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line. The invention also provides a method of purifying the CSF proteins and the purified proteins so produced.

617042

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