Preparation and use of ion-exchange chromatographic supports based on perfluoropolymers.
A poly(vinyl alcohol) (PVA) coated particulate perfluoropolymer (FEP) support has been functionalised with ion-exchange groups for use in ion-exchange chromatography of proteins. Anion-exchange (DEAE and Q) and cation-exchange (SP) groups were introduced to PVA-FEP which had previously been activated using cyanuric chloride. The equilibrium adsorption capacities of SP-PVA-FEP were 31.8 and 25.2 mg ml-1 for lysozyme and IgG respectively while for DEAE-PVA-FEP, the equilibrium adsorption capacities were 14.9 and 9.7 mg ml-1 for beta-lactoglobulin and HSA respectively. The equilibrium adsorption capacities for Q-PVA-FEP were determined to be 17.2 and 13.5 mg ml-1 for beta-lactoglobulin and HSA respectively. Experiments carried out to investigate the resolving power of materials showed that both SP and Q-PVA-FEP were able to resolve proteins with only small differences in their isoelectric points and that this resolution could be maintained at a flow-rate of 1500 cm h-1. SP-PVA-FEP was used to purify lysozyme from egg whites where a 50-fold purification, to homogeneity, was achieved in 98% yield. The anion exchanger, Q-PVA-FEP could be used to purify G6PDH from a clarified homogenate of bakers' yeast 14.3-fold in 81% yield.
McCreath
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GE
Department of Chemical Engineering, University of Cambridge, UK.
Owen
R O
RO
Nash
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DC
Chase
H A
HA
D000327:Adsorption; D000838:Anions; D002412:Cations; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004529:Egg White / Q000032:analysis; D017611:Fluorine Compounds / Q000737:chemistry; D005954:Glucosephosphate Dehydrogenase / Q000302:isolation & purification; D007475:Ion Exchange Resins / Q000737:chemistry; D007526:Isoelectric Point; D015394:Molecular Structure; D009113:Muramidase / Q000302:isolation & purification; D011108:Polymers / Q000737:chemistry; D011142:Polyvinyl Alcohol; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D014227:Triazines
D000838:Anions; D002412:Cations; D017611:Fluorine Compounds; D007475:Ion Exchange Resins; D011108:Polymers; D014227:Triazines; C019309:cyanuric chloride; D011142:Polyvinyl Alcohol; D005954:Glucosephosphate Dehydrogenase; D009113:Muramidase