The chromatography of serum proteins on cellulose ion exchange columns

The chromatography of normal serum proteins

“…Similar results have been obtained with carboxymethylcellulose (Cooke, Tombs, Souter, Weston & Maclagan, 1959). The detailed electrophoretic analysis of chromatographic fractions (Fahey et al 1958) has shown that, whereas both the a-and f-globuilins are resolved into a number of subfractions on diethylaminoethylcellulose columns, the ax-globulin subfractions are not separated from those of the f,globulins.…”

“…Although one of the main advantages of ionexchange cellulose chromatography is the large amount of protein which can be handled by comparatively small columns, very small amounts of protein (less than 1 mg.) could also be successfully run on DEAE-cellulose. Although excessive trailing prevented satisfactory results on CMcellulose, patterns similar to normal CM-cellulose runs could be obtained with cellulose phosphate on a small scale (Cooke et at. 1959).…”

“…Ammonium sulphate fractionation is one such method but, as with column chromatography, its resolving power is poor for a-and ,B globulins. A combination of these two methods gave good results with the more soluble proteins (Cooke et al 1959;Tombs, James & Maclagan, 1961) but was not applicable to the remainder. Altemating chromatography on CMand DEAE-cellulose has been used for fine resolution of y-globulin (Sober & Peterson, 1958).…”

“…At 280 mju an automatic scanner measuring continuously on the column effluent was used. This apparatus is a development of that described by Cooke et al (1959) and will be reported in detail elsewhere.…”

“…The detailed electrophoretic analysis of chromatographic fractions (Fahey et al 1958) has shown that, whereas both the a-and f-globuilins are resolved into a number of subfractions on diethylaminoethylcellulose columns, the ax-globulin subfractions are not separated from those of the f,globulins. Cooke et al (1959) showed the utility of ammonium sulphate fractionation followed by diethylaminoethylcellulose chromatography for separation of the more soluble proteins, but indicated that the less soluble a-and fl-globulins were poorly resolved. Alternating chromatography on carboxymethyland diethylaminoethyl-cellulose has found some application, but suffers from practical difficulties which restrict its use for serum protein.…”