	US 12,331,335 B2
	Methods for production of human recombinant arginase 1 and uses thereof
Scott W. Rowlinson, Austin, TX (US)
Assigned to Immedica Pharma AB, Stockholm (SE)
Filed by Immedica Pharma AB, Stockholm (SE)
Filed on Jun. 20, 2023, as Appl. No. 18/338,255.
Application 18/338,255 is a continuation of application No. 17/006,383, filed on Aug. 28, 2020, abandoned.
Claims priority of provisional application 62/894,319, filed on Aug. 30, 2019.
Prior Publication US 2024/0110172 A1, Apr. 4, 2024
Int. Cl. C12N 9/78 (2006.01); C07K 1/18 (2006.01); C07K 1/36 (2006.01); C12N 1/06 (2006.01)

CPC C12N 9/78 (2013.01) [C07K 1/18 (2013.01); C07K 1/36 (2013.01); C12N 1/06 (2013.01); C12Y 305/03001 (2013.01)]

16 Claims

1. A method for producing purified recombinant cobalt-substituted human Arginase, wherein the recombinant human Arginase (rhARG) comprises an amino acid sequence that is at least 98% identical to SEQ ID NO: 1, the method comprising:

a) in a bioreactor, culturing E. coli cells that produce rhARG;

b) lysing the E. coli cells;

c) removing cell debris from the lysate;

d) loading the cell lysate on a cation exchange column;

e) eluting the rhARG with a high salt solution;

f) incubating the eluted rhARG with a cobalt salt to form cobalt-substituted rhARG (Co-rhARG);

g) applying said Co-rhARG to an anion exchange column and collecting the flow-through;

h) applying the flow-through onto a third chromatography column; and

i) eluting the Co-rhARG from the third chromatography column with a high salt solution.