Methods of purification of cytochrome P450 proteins
OwnerAlison Ward, Jose Cosme +3inventors
InventorsAlison Ward, Jose Cosme, Pamela Williams, Bruce Hamilton, Laurent Vuillard
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Abstract
The invention provides a method for the purification of cytochrome P450 molecules, the method comprising expressing in a host cell culture a cytochrome P450 molecule; recovering said cells from said culture and suspending said cells in a high salt buffer; lysing said cells and removing cell debris to provide a high-salt lysate; adding to said lysate a detergent to provide a high-salt-detergent lysate; and recovering said P450 from said lysate. The method provides yields of P450 proteins suitable for crystallization.
Claims13
A method for the purification of a cytochrome P450, wherein said method comprises:
- (a) expressing in a host cell culture a cytochrome P450 molecule;
- (b) recovering said cells from said culture and suspending said cells in a salt buffer having a conductivity of from 12 to 110 mS/cm;
- (c) lysing said cells and removing cell debris to provide a high-salt lysate;
- (d) adding to said lysate a detergent to provide a high-salt-detergent lysate;
- and
- (e) recovering said P450 from said lysate.
13-20. (canceled)
Dependencies are inferred from the claim text.