Aventis Pharma Deutschland, Gmbh

Frank Richard

Reinhold Keller

Richard Bicker

Werner Sievers

J&ouml;rg Von Eysmondt

Dieter Desch

Jorg Moller

The present invention relates to an improved procedure for the chromatographic purification of insulins wherein a pressure-stable organic polymeric chromatography material is used as a stationary phase, and the mobile phase contains at least one water-miscible organic solvent and at least one buffer substance and the pH is from about 7 to about 11.

Procedure for the chromatographic purification of insulins

A peptide such as in a cell-free fermentation medium is ultrafiltered with a membrane having a molecular weight cut-off of at least approximately two times greater than the molecular weight of the peptide. The peptide is retained by the membrane, and is desalinated and concentrated to provide prepurification of the peptide. A preferred temperature for ultrafiltration is about 5 to about 15.degree. C. Permeate flow rate may be about 10 l/m.sup.2 /h to about 35 l/m.sup.2 /h, and permeate conductivity may be less than about 10 mS/cm. The permeate may be recycled until peptide concentration is constant. A cell-free fermentation medium containing hirudin from a recombinant microorganism such as Saccharomyces cerevisiae is ultrafiltered with a membrane having a molecular weight cut-off of about 20 kD to about 30 kD.

Ultrafiltration process for desalination and concentration of a peptide

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Kronberg, Germany

Frank Richard