Biospectrum, Inc.

Marian Kruzel

Jerzy Alexander Georgiades

A process for the purification of human interferon alpha involving a three-step chromatography sequence under highly dissociating conditions is disclosed. The purification method involves a first step of loading crude interferon alpha onto a glass sorbent column to adsorb interferon alpha. The adsorbed interferon alpha is then eluted from the glass sorbent using a hydrophobic electrolyte solution comprising tetramethyl ammonium chloride, NaCl and propylene glycol. Thereafter, in a second step, the eluted interferon is subjected to SEPHACRYL S-200 molecular sieving chromatography under highly dissociating conditions and the fractions corresponding to molecular weights ranging from about 10,000 to about 40,000 are collected. In a third step, the collected fractions are then passed thorugh a column Zn++ chelating resin. The interferon alpha passes through unadsorbed to the Zn++ chelating resin. The resulting solution is a highly purified interferon alpha which preserves the apparently native isospecies compositions. Also disclosed is a method to enhance overall interferon alpha recovery which involves treating crude or partially purified interferon alpha solutions with a non-ionic detergent such as, for example, TRITON X-100, TWEEN 20, or NONIDET P-40.

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Houston, TX, United States of America

Marian Kruzel