title: Protein-selective coacervation with hyaluronic acid.
abstract: Selective coacervation with hyaluronic acid (HA), a biocompatible and injectable anionic polysaccharide, was used to isolate a target protein, bovine serum albumin (BSA), with 90% purity from a 1:1 mixture with a second protein of similar pI, β-lactoglobulin (BLG). This separation was attributed to the higher HA-affinity of BSA, arising from its more concentrated positive domain. The values of pH corresponding respectively to the onset of complex formation, coacervation, precipitation, and redissolution (pH(c), pHϕ, pH(p), and pH(d)) were determined as a function of ionic strength I. These pH values were related to critical values of protein charge, Z, and their dependence on I provided some insights into the mechanisms of these transitions. The higher polyanion binding affinity of BSA, deduced from its higher values of pH(c), was confirmed by isothermal titration calorimetry (ITC). Confocal laser microscopy clearly showed time-dependent coalescence of vesicular droplets into a continuous film. Comparisons with prior results for the polycation poly(diallyldimethylammonium chloride) (PDADMAC) show reversal of protein selectivity due to reversal of the polyelectrolyte charge. Stronger binding of both proteins to PDADMAC established by ITC may be related to the higher chain flexibility and effective linear charge density of this polycation.
authors: Department of Polymer Science and Engineering and ‡Department of Chemistry University of Massachusetts , Amherst, Massachusetts, 01003, United States.
authors: Xiaosong
authors: X
authors: Du
authors: Paul L
authors: PL
authors: Dubin
authors: David A
authors: DA
authors: Hoagland
authors: Lianhong
authors: L
authors: Sun
mesh_terms: D000818:Animals; D000838:Anions / Q000737:chemistry; D002417:Cattle; D006820:Hyaluronic Acid / Q000737:chemistry*; D007782:Lactoglobulins / Q000737:chemistry* / Q000302:isolation & purification; D011095:Polyethylenes / Q000737:chemistry; D011134:Polysaccharides / Q000737:chemistry; D000644:Quaternary Ammonium Compounds / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D000838:Anions; D007782:Lactoglobulins; D011095:Polyethylenes; D011134:Polysaccharides; D000644:Quaternary Ammonium Compounds; C041004:poly-N,N-dimethyl-N,N-diallylammonium chloride; D012710:Serum Albumin, Bovine; D006820:Hyaluronic Acid