AFFINITY-CHROMATOGRAPHY ON NOVEL PERFLUOROCARBON SUPPORTS - IMMOBILIZATION OF CI REACTIVE BLUE-2 ON A POLYVINYL ALCOHOL-COATED PERFLUOROPOLYMER SUPPORT AND ITS APPLICATION IN AFFINITY-CHROMATOGRAPHY

The triazine dye C.I. Reactive Blue 2 has been immobilised on a particulate perfluorocarbon support by preparation of a hydrophilic polymeric coating comprising polyvinyl alcohol (average molecular weight, Mr 14 000) esterified with perfluorooctanolyl chloride and securely adsorbed ont he perfluorocarbon support by multiple Van der Waals interactions. This polyvinyl alcohol-based coating wets the perfluorocarbon support and provides a neutral barrier to non-specific adsorption of proteins. Reaction with the triazine dye C.I. Reactive Blue 2 allows secure immobilisation of this versatile pseudo-affinity ligand and yields a remarkably stable adsorbent. The performance and capacity of the perfluorocarbon-based adsorbent was assessed: an 8-fold purification of lactable dehydrogenase was achieved from a crude rabbit muscle extract in 100% yield. The albumin-binding capacity of a dyed perfluorocarbon support containing 2.2±0.2 μmol dye per g wet support was determined to be 15.7 mg/ml by frontal analysis. These supports compare favourably with conventional polysaccharidic affinity supports. © 1990.