ONE-STEP CHROMATOGRAPHIC ISOLATION OF ORGOTEIN FROM RED BLOOD CELLS

摘要

1386453 Orgotein DIAGNOSTIC DATA Inc 6 Dec 1972 [7 Dec 1971] 56243/72 Heading C3H A process for the isolation of orgotein from red blood cells involves subjecting a mixture of buffer-soluble red blood cell proteins which mixture contains at least orgotein, haemoglobin and carbonic anhydrase, to a chromatographic separation by (a) contacting the mixture of buffer-soluble proteins as an aqueous solution having an ionic strength of less than 0À01 M with an ion exchange resin having weakly basic groups, thereby adsorbing the orgotein on the resin (b) washing the ion exchange resin free of haemoglobin (c) eluting the adsorbed orgotein from the resin with an aqueous medium having a higher ionic strength and at a pH of approximately 6 and (d) separating orgotein from the fraction of the eluate having a peak divalent metal content (copper and zinc). Substantially pure orgotein is typically eluated at ionic strengths in the range 0À02-0À035 M. The isolated orgotein may be further purified by gel filtration or other conventional techniques. The preferred ion exchange resins are tertiary or quaternary amino-celluloses or -dextrans (specified). The starting mixture may be obtained from bovine, rabbit, monkey, rat, chicken or human red blood cells.