Polyelectrolyte-coated ion exchangers for cell-resistant expanded bed adsorption.
Adsorption chromatography in expanded beds is a widely used technology for direct capture of target proteins from fermentation broths. However, in many cases this method cannot be applied as a result of the strong tendency of cells or cell debris to interact with the adsorbent beads. To prevent contamination of the expanded bed with the biomass, STREAMLINE DEAE, anion exchanger designed for expanded bed adsorption, was modified with a layer of poly(acrylic acid) (PAA). The shielding layer of polyelectrolyte was attached to the surface of the matrix beads via electrostatic interactions. PAA with a high degree of polymerization was chosen to prevent diffusion of large polymer molecules into the pores of adsorbent. Thus, the shielding layer of PAA was adsorbed only at the mouth of the pores of STREAMLINE DEAE beads and only marginally decreased the binding capacity of the ion exchanger for bovine serum albumin, the model protein in this study. PAA-coated STREAMLINE DEAE practically did not interact with yeast cells, which otherwise bound strongly to the native adsorbent at neutral conditions. Cell-resistant PAA-coated anion exchanger was successfully used for isolation of BSA from the model protein mixture containing BSA, lysozyme (positively charged at applied conditions), and yeast cells. The layer of PAA was stable under mild elution conditions, and the modified adsorbent could be used in the repeated purification cycles.
Dainiak
Maria B
MB
Department of Biotechnology, Center for Chemistry and Chemical Engineering, Lund University, P.O. 124, S-221 00 Lund, Sweden.
Galaev
Igor Yu
IY
Mattiasson
Bo
B
D000327:Adsorption; D000837:Anion Exchange Resins / Q000737:chemistry*; D018533:Biomass; D002448:Cell Adhesion; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods*; D004573:Electrolytes / Q000737:chemistry*; D004983:Ethanolamines / Q000737:chemistry; D005285:Fermentation; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification; D012685:Sepharose / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D012996:Solutions / Q000737:chemistry; D055672:Static Electricity; D013997:Time Factors; D015003:Yeasts / Q000166:cytology
D000837:Anion Exchange Resins; D004573:Electrolytes; D004983:Ethanolamines; D012996:Solutions; D012710:Serum Albumin, Bovine; D012685:Sepharose; D009113:Muramidase; C007369:2-diethylaminoethanol