Patent ID: 9029331
Filing Date: 2015-05-12
Classification: A61K,A61P,C07K

Abstract:
1. A method of preparing a purified, pegylated GCSF peptide, which method comprises (a) providing a composition comprising water, a buffer, at least one polysorbate, and 20 mM L-methionine, wherein the polysorbate is polyoxyethylene sorbitan monolaurate or polyoxvethylene sorbitan monooleate, and the composition has a pH of about 6.5; (b) combining the composition of step (a) with reagents comprising GCSF, GalNAeT2, and UDP-GalNAc, wherein the composition of step (b) comprises 0.45 mM UDP-GalNAc; (c) combining the composition of step (b) with MnCl (d) combining the composition of step (c) with ST6GalNAc1 and an aliquot of cytidine monophosphate sialic acid poly(ethylene glycol) (CMP-SA-PEG), wherein the composition of step (d) comprises 1 mM CMP-SA-PEG, and, wherein the composition of step (e) is not purified prior to step (d); (e) combining the composition of step (d) with an aliquot of CMP-SA-PEG, wherein the composition of step (e) comprises 1.5 mM CMP-SA-PEG; (f) combining the composition of step (e) with a 20 mM citrate buffer, wherein no additional L-methionine is added to the composition of step (e) and wherein the resulting composition has a pH of about 4, and (g) applying the composition of step (f) to a cation exchange chromatography column, whereby purified PEG-GCSF is produced.