Purifying equine interferon beta - using anion and cation-exchange columns, useful for antiviral or immuno:modulatory medicament

摘要

A method for purifying equine interferon beta (Eq IFN-beta) comprises: (a) inactivating E. coli host cells with acid and extracting the EqIFN-beta with an alkaline medium. (b) precipitating the cell culture supernatant with polyethylamine and running this on an ion-exchange column and/or a molecular sieve. (c) eluting the EqIFN-beta with a phosphate buffer; and (d) purifying the eluted interferon further using an anion and/or cation exchanger. The extraction pH is 10 and in stage (b), adsorption on the column is achieved by the use of hydroxyapatite. The anion-exchanger is monoQ and the cation-exchanger is Mino S. Elution on the anion exchange column uses a linear gradient of sodium chloride. For the cation exchange column, a linear gradient of ammonium sulphate is used. USE/ADVANTAGE - The purified EqIFN-beta is useful as an antiviral medicament and also has immunomodulatory activity.