Novel affinity separations based on perfluorocarbon emulsions. Use of a perfluorocarbon affinity emulsion for the purification of human serum albumin from blood plasma in a fluidised bed.
A perfluorocarbon affinity emulsion has been generated by homogenisation of a saturated perfluorocarbon oil with a polymeric fluorosurfactant based on poly(vinyl alcohol) (relative molecular mass 9000-10,000) previously derivatised with the triazine dye CI Reactive Blue 4. This affinity emulsion has subsequently been cross-linked in situ and used in a fluidised bed for the purification of human serum albumin (HSA) from blood plasma. HSA was quantitatively recovered in a semi-continuous fashion from plasma at an average purity of 90 +/- 3.3%. The albumin binding capacity of the emulsion has been shown to be 0.59 mg/ml by frontal analysis corresponding to a mol/mol ligand usage of 13.5%. In all regards, when used in a fluidised bed, the emulsions have been shown to behave as a normal chromatographic material. They are stable under operational conditions with no coalescence being observed for periods greater than 1 year. These novel liquid affinity supports present an exciting opportunity to develop a range of unit operations for the continuous purification of proteins.
McCreath
G E
GE
Department of Chemical Engineering, University of Cambridge, UK.
Chase
H A
HA
Purvis
D R
DR
Lowe
C R
CR
D002846:Chromatography, Affinity / Q000379:methods*; D002850:Chromatography, Gel; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D004655:Emulsions; D005466:Fluorocarbons; D006801:Humans; D008024:Ligands; D008970:Molecular Weight; D012709:Serum Albumin / Q000302:isolation & purification*; D013501:Surface-Active Agents; D014227:Triazines
D003432:Cross-Linking Reagents; D004655:Emulsions; D005466:Fluorocarbons; D008024:Ligands; D012709:Serum Albumin; D013501:Surface-Active Agents; D014227:Triazines; C036598:procion blue MX-R