Chromatographic properties of gamma globulin: behavior of serum gamma macroglobulins.

The cellulose ion exchangers (1) have proved to be useful chromatographic media for the serum proteins. In the fractionation of normal human serum on the anion exchanger, diethylaminoethyl (DEAE) cellulose, application of buffer gradients of increasing concentration and decreasing pH results in the appearance of components in the effluent fluid in order of increasing electrophoretic mobility (2), indicating that ion exchange is the predominant mechanism involved. In the case of the gamma globulin component, some subfractionation was demonstrated in terms of progressively increasing electrophoretic mobility of column effluents of this fraction. Little attention has been directed, however, to the chromatographic properties of the gamma globulins of high molecular weight, 19S or greater, which are found in large quantities in certain pathological sera, and also in small quantities in normal human sera (3). In the course of anion exchange fractionation of sera from patients with rheumatoid arthritis, the "rheumatoid factor" (4-6), a gamma globulin constituent of high molecular weight (7), appeared in column effluents far removed from the bulk of the gamma globulin (8). This observation indicated that the chromatographic behavior of certain gamma globulins, and possibly of other serum proteins, on DEAE cellulose is governed not only by ion exchange but also by other factors which depend in part upon molecular size. Our attention was therefore directed to sera known to