title: Separation of macromolecules by ultrafiltration: removal of poly(ethylene glycol) from human albumin.
abstract: Mixtures of albumin and poly(ethylene glycol) (PEG) were used to elucidate some of the factors which influence the separation of macromolecules by thin-channel ultrafiltration. Several membranes which readily passed PEG-4000 in the absence of protein were found to exhibit increased rejection of the synthetic polymer when albumin was added to the system. Based on a comparison of filtration flux and net sieving properties, the PM-30 membrane of Amicon was chosen for further characterization. The increased rejection of PEG-4000 in the presence of albumin was independent of albumin concentration between 1 and 100 mg/ml and persisted even after albumin was removed and the system flushed with water. Overnight incubation of the membrane with trypsin restored the original sieving properties, indicating that the 'permanent' effects were due to irreversible adsorption to the membrane. By measuring flux over a 10(6)-fold range of albumin concentration it was possible to resolve the effects of protein adsorption, a saturable process which occurs at low protein concentration (< 0.01 mg/ml), from the effects of concentration polarization which occur at high protein concentration (> 0.1 mg/ml). Only the former process has an effect on the net sieving properties in this system. In spite of the adverse effects of protein adsorption, it was still possible to obtain efficient removal of PEG-4000 from albumin. Exchange of approximately 5 vols. of solvent at room temperature resulted in a 10-fold reduction in the concentration of PEG in the sample, with no loss of albumin, and no formation of albumin dimers.
mesh_terms: D000327:Adsorption; D006801:Humans; D011092:Polyethylene Glycols / Q000302:isolation & purification*; D012709:Serum Albumin / Q000302:isolation & purification*; D014462:Ultrafiltration / Q000379:methods
chemical_list: D012709:Serum Albumin; D011092:Polyethylene Glycols