Direct purification of lysozyme using continuous counter-current expanded bed adsorption.
We describe the application of a novel technique for the continuous counter-current chromatography of proteins. The unit operation has shown potential in extracting targeted species from unclarified feedstocks, delivering clarified streams of purified product. The adsorbent used in this equipment consists of a perfluorocarbon matrix, coated with poly(vinyl alcohol), and derivatized with the triazine dye Procion Red HE-7B. Purification of lysozyme from egg-whites and enriched bovine milk could be carried out continuously. The former was extracted in 90.5% yield at a rate of 7400 U/min, achieving a purification factor of 19.4. Lysozyme from the enriched milk sample was extracted continuously at a rate of 41000 U/min, in 66.0% yield. The continuous products streams in both cases were fully clarified, thus enabling their direct application to a final polishing step, if desired.
Owen
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Department of Chemical Engineering, University of Cambridge, UK.
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D000818:Animals; D002417:Cattle; D002645:Chickens; D004396:Coloring Agents / Q000737:chemistry; D003377:Countercurrent Distribution / Q000379:methods*; D004527:Egg Proteins / Q000737:chemistry*; D005465:Fluorocarbon Polymers / Q000737:chemistry; D008892:Milk / Q000737:chemistry* / Q000201:enzymology; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D014227:Triazines / Q000737:chemistry
D004396:Coloring Agents; D004527:Egg Proteins; D005465:Fluorocarbon Polymers; D014227:Triazines; C105892:Procion Red HE-7B; D009113:Muramidase