US 12,391,728 B2
Full flow-through process for purifying recombinant proteins
Didier Duthe, Paris (FR); Céline Hemet, Paris (FR); Benoit Mothes, Paris (FR); and Jérome Pezzini, Paris (FR)
Assigned to SANOFI, Paris (FR)
Appl. No. 17/040,702
Filed by SANOFI, Paris (FR)
PCT Filed Mar. 27, 2019, PCT No. PCT/EP2019/057687
§ 371(c)(1), (2) Date Sep. 23, 2020,
PCT Pub. No. WO2019/185691, PCT Pub. Date Oct. 3, 2019.
Claims priority of application No. 18305338 (EP), filed on Mar. 27, 2018.
Prior Publication US 2021/0009634 A1, Jan. 14, 2021
Int. Cl. A61K 39/395 (2006.01); C07K 1/36 (2006.01); C07K 16/00 (2006.01)
CPC C07K 1/36 (2013.01) [C07K 16/00 (2013.01); C07K 2317/24 (2013.01)] 20 Claims
 
1. A full flow-through method for purifying a protein from solution comprising:
(a) a filtration step comprising:
passing said solution over at least one chelating agent matrix in the flow-through mode, wherein said at least one chelating agent matrix is activated carbons,
recovering a filtered protein solution from the flow-through of said at least one chelating agent matrix;
(b) an exchanging step comprising:
passing the filtered protein solution obtained at the end of step (a) over at least one diafiltration membrane using only one buffer for the exchange,
recovering the partly purified protein-containing retentate of said at least one diafiltration membrane; and
(c) a polishing step comprising:
passing the retentate obtained at the end of step (b) over a combination of membrane adsorbers in the flow-through mode, wherein two membrane adsorbers of said combination of membrane adsorbers are orthogonal in terms of mechanisms of action, and said combination of membrane adsorbers has been equilibrated beforehand with an equilibration buffer which is identical to the one buffer used for exchange at step (b),
recovering purified protein from the flow-through of said combination of membrane adsorbers;
wherein said purifying method does not include a Protein A chromatography step.