Process for the production of purified cytidinemonophosphate-sialic acid-polyalkylene oxide (CMP-SA-PEG) as modified nucleotide sugars via anion exchange chromatography

摘要

The current invention provides methods (e.g., large-scale processes) for the production of nucleotide sugars, which are modified with a polymeric modifying group, such as poly(alkylene oxide) moieties (e.g., poly(ethylene glycol) or poly(propylene glycol)) moieties. A typical process of the invention includes anion exchange chromatography followed by an ultrafiltration procedure, such as tangential flow filtration. The process of the invention provides modified nucleotide sugars in unexpectedly high purity and high overall yields.

主权项

1. A method of making a composition comprising a modified nucleotide sugar covalently linked to a polymeric modifying group, said polymeric modifying group comprising at least one linear or branched poly(alkylene oxide) moiety, said method comprising:
(i) contacting a reaction mixture comprising said modified nucleotide sugar with a Q-sepharose anion exchange resin; (ii) eluting said modified nucleotide sugar from said Q-sepharose anion exchange resin using a bicarbonate buffer, wherein said eluting comprises a first step wherein said buffer comprises between about 0.01 mM and about 10 mM of bicarbonate and a second step wherein said buffer comprises between about 10 mM and about 30 mM of bicarbonate, thereby forming an eluate fraction comprising said modified nucleotide sugar; and (iii) desalting said eluate fraction, wherein said method does not comprise ultrafiltration prior to step (i), thereby forming said composition, wherein said modified nucleotide sugar comprises cytidine-monophospho-sialic acid (CMP-SA), wherein said method provides said modified nucleotide sugar in a purity between about 80% and about 100% (w/w) and wherein the linear or branched poly(alkylene oxide) moiety has a molecular weight in the range of 10-40 kDa.