Chromatographic Studies of the Rheumatoid Factor

Abstract

Since the discovery that sera from patients with rheumatoid arthritis have the capacity to agglutinate sensitized sheep erythrocytes (1), much attention has been directed to the constituents in serum responsible for this phenomenon. This has resulted not only in the development of a number of diagnostic tests, but also in considerable investigation (2) into the nature of the responsible serum constituent. The latter has been referred to as the rheumatoid factor (3).Ziff and coworkers (4) demonstrated that the agglutinating activity of rheumatoid serum is precipitated in the euglobulin fraction, and Svartz and Schlossmann (5) showed that it precipitated in the "cold globulin" fraction. On zone electrophoresis of the cold globulin precipitate, agglutinating activity was found in the gamma globulin. This has also been demonstrated by paper electrophoresis of precipitated material obtained after dilution of rheumatoid serum with water (6). Ultracentrifugal studies of rheumatoid sera and euglobulin fractions have shown that the factor circulates as a high molecular weight component with a sedimentation coefficient of 22 S (7), and that there is a correlation between the quantity of the 22 S component found and the agglutination titer.With the development of a suitable chromatographic system for the fractionation of serum proteins, it has been possible to apply the chromatographic method to the investigation of the rheumatoid factor. The cellulose ion exchangers described by Sober and Peterson (8,9) have therefore been employed by , and by Svartz et al. (13), for the fractionation of rheumatoid serum and euglobnlin fractions. The present paper describes the isolation of rheumatoid factor using an anion exchanger and further fractionation of the purified material on a cation exchanger.

Separation of rheumatoid factors of different specificities using columns conjugated with gamma‐globulin

“…Evidence for additional anti-7-globulin factors in rheumatoid arthritis sera which react only with enzymaticallydegraded antibody, has been obtained recently. 23 The hypothesis of multiple types of antibodies to buried determinants of 7 …”

Role of igg and igm antinuclear antibodies in formation of lupus erythematosus cells and extracellular material

“…After 48 hr of dialysis against 0.011M sodium phosphate buffer, p H 7.0, at 4" C, sera were passed through a DEAEcellulose (Whatman DE-32) anion exchange column by a three-step elution technic with successive posphate buffers: 0.01M, pH 7.0; O.lOM, pH G.0; and O.SOM, p H 5.0 (14). The first peak eluted contained only IgG by immunodiffusion; the second and third peaks contained mostly IgA and IgM, respectively.…”

The Formation of Macroglobulin Antibodies. I. Studies on Adult Humans*

“…The fractions comprising each peak were pooled and tested for antibodies to typhoid 0, typhoid H, and paratyphoids A and B by standard agglutination procedures (10). Antibody activities associated with the first (pH 7.0, 0.01 M) and the last (pH 5.0, 0.3 M) chromatographic peaks were considered to contain 7S and 19S -y-globulins, respectively, on the basis of the behavior of these proteins on DEAE-cellulose (9,11,12). Although the first, or 7S, peak contains no 19S globulins, the last peak is contaminated by 7S protein, which could account for part of the antibody titers associated with this fraction.…”

“…Chromatographic separation was carried out on diethylaminoethyl (DEAE)-cellulose columns as described previously (9). The fractions comprising each peak were pooled and tested for antibodies to typhoid 0, typhoid H, and paratyphoids A and B by standard agglutination procedures (10).…”