Patent ID: 6433144
Filing Date: 2002-08-13
Classification: A61K,C07K

Abstract:
A method of purifying Type I interferons from a mixture containing Type I interferons, human serum albumin and other trace proteins comprising:a) abstaining from use of immunoaffinity chromatography and applying said mixture to cation exchange chromatography and obtaining a pool of crude interferon and adjusting said pool of crude interferon to a pH of 6.5; b) subjecting said pool of crude interferon to a hydroxyapetite column (HAC); c) eluting a first eluate from said HAC column by use of a first eluant having a pH of about 5; d) applying said first eluate containing natural human Type I interferons to a size exclusion chromatography column under dissociating, non-denaturing conditions, said conditions comprising about 50 mM NaCl and about 25-75 mM tris hydroxymethyl aminomethane hydrochloride, pH 7.4, wherein said size exclusion chromatography column separates proteins of size range from 3 kDa to about 70 kDa; and e) resolving the size exclusion chromatogram with a resolving buffer having a pH below 7.4 until said human serum albumin and Type I interferon have been eluted from the column and collected separately.