Method of producing a recombinant protein
The present invention relates to a method of producing a recombinant protein in a host cell comprising adding Polyethyleneimine (PEI) during cell culture. Addition of PEI to the cell culture as a fermentation enhancer can result in reducing the viscosity of the cell culture, and/or increasing the extracellular concentration of the recombinant protein, and/or reducing the duration of cell culture to the point of harvest or protein recovery.
1. A method of producing a recombinant protein in a host cell comprising providing a cell culture, inducing expression of the recombinant protein, adding polyethyleneimine to said cell culture, and harvesting the recombinant protein directly from an extracellular medium of the cell culture without pre-treatment of the cell culture by thermal, mechanical, or chemical lysis, thereby producing the recombinant protein, wherein polyethyleneimine is added at any point in time from induction of expression of the recombinant protein to 30 minutes prior to harvest.
2. The method according to claim 1 , wherein the host cell or cell culture is a Gram-negative bacteria with a periplasm.
3. The method according to claim 2 wherein the Gram-negative bacteria is Escherichia coli.
4. The method according to claim 1 , wherein polyethyleneimine is added 30 minutes prior to harvest.
5. The method according to claim 1 , wherein polyethyleneimine is added at the time of induction of expression of the recombinant protein.
6. The method according to claim 1 , wherein the polyethyleneimine is added continuously during cell culture.
7. The method according to claim 1 , wherein the polyethyleneimine is branched.
8. The method according to claim 1 , wherein the molecular weight of the polyethyleneimine is from 50-1000 kDa.
9. The method according to claim 8 , wherein the molecular weight of the polyethyleneimine is about 750 kDa.
10. The method according to claim 1 , wherein the recombinant protein (i) is an antigen binding protein; and/or (ii) has a Molecular Weight (MW) of 25 kDa or less.
11. The method according to claim 10 , wherein the antigen binding protein is an antibody.
12. The method according to claim 10 , wherein the antigen binding protein is a domain antibody (dAb).
13. The method according to claim 1 , further comprising purifying the harvested recombinant protein.
14. The method according to claim 13 , wherein purifying the harvested recombinant protein comprises one or more chromatography steps.