Chromatographic isolation of proteins from e.g. blood esp. cytochrome C sepn. - by contacting with buffered macroporous cellulose@ beads, rinsing away impurities and eluting adsorbed proteins with different buffer

摘要

Chromatographic isolation of proteins is effected by (a) contacting a protein-contg. aq. soln. with macroporous cellulose beads at a pH of at least 7, a buffer concn. of up to 0.1M and a temp. of 0-30 deg. C; (b) removing any impurities by washing the beads; and (c) eluting the adsorbed protein(s) with a different buffer soln. USE - Used to isolate blood or serum proteins, esp. to separate cytochrome C form haemoglobin and/or myoglobin, to separate ferritin from cytochrome C, haemoglobin and/or myoglobin, to separate serum albumin from haemoglobin or to remove gamma-globulins from serum, or to isolate lysozyme from solns. In an example, a soln of cytochrome C in buffer A (0.01M borate, pH9) was applied to a column contg. 10ml of preswollen macroporous cellulose beads equilibrated with buffer A. The column was washed with buffer A and eluted with 0.1M phosphate buffer (pH 7). The cytochrome C eluted as a sharp band and could be isolated in pure form from the elu