Purification of hepatitis proteins

Details Purification of hepatitis proteins Purification of hepatitis proteins

C - Chemistry, Metallurgy

07

K

167/134, 530/15.

C07K 14/02 (2006.01) C07K 1/36 (2006.01)

Patent

CA 1339792

The present invention pertains to a process for recovering hepatitis B surface antigen from recombinant Pichia pastoris cells comprising: a) lysing said yeast cells in the presence of a buffered chaotropic salt and separating the hepatitis B surface antigen-containing supernatant from said lysed cells; b) subjecting the hepatitis B surface antigen-containing supernatant obtained in step (a) to conditions suitable to precipitate lipids and contaminant proteins from said supernatant; c) subjecting the hepatitis B surface antigen-containing supernatant obtained in step (b) to diafiltration; d) contacting the hepatitis B surface antigen-containing retentate obtained in step (c) with silica; e) washing contaminant proteins from the resulting silica-adsorbed hepatitis B surface antigen with a buffer having a pH within the range of 6-8; f) eluting the hepatitis B surface antigen from the silica with a buffered eluant having a pH within the range of 9.5-11.0 containing from 0.5 to 8 molarity of urea; g) subjecting the hepatitis B surface antigen-containing fraction obtained in step (f) to a gel filtration step suitable to separate said hepatitis B surface antigen from contaminant proteins; and h) subjecting the hepatitis B surface antigen-containing fraction obtained in step (g) to an anion exchange chromatography step suitable to separate said hepatitis B surface antigen from contaminant proteins.

596774

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