LOSPALLUTO J, ZIFF M
J Exp Med. 1959 Aug 1;110(2):169-86. doi: 10.1084/jem.110.2.169.
Serum and serum fractions from patients with rheumatoid arthritis have been subjected to anion exchange chromatography on diethylaminoethyl cellulose. The rheumatoid factor activity was clearly separated from the bulk of the gamma globulin by virtue of the anomalous behavior of macroglobulins on this column. Purification of rheumatoid factor by precipitation with fraction II, resolution in 4 M urea, and two successive fractionations on an anion exchanger yielded a highly active preparation consisting of approximately 95 per cent 19S gamma globulin with no detectable amount of 7S contaminant. Evidence was obtained for the existence of two factors by cation exchange chromatography (carboxymethyl cellulose) of rheumatoid serum and of macroglobulin previously separated by anion exchange chromatography. One factor (factor I) agglutinated sensitized sheep cells and latex particles and also precipitated with human fraction II. The second (factor II) agglutinated latex partides and precipitated with fraction II but did not agglutinate sensitized sheep cells. A substance with the properties of factor II was demonstrated in two of four normal sera by cation exchange chromatography. It was suggested that the agglutination of latex particles and precipitation with human fraction II, observed in the case of certain abnormal sera, may be due to elevated concentrations of factor II.
类风湿性关节炎患者的血清及血清组分已在二乙氨基乙基纤维素上进行阴离子交换层析。类风湿因子活性因巨球蛋白在此柱上的异常行为而与大部分γ球蛋白明显分离。通过用II组分沉淀、在4M尿素中溶解以及在阴离子交换剂上连续两次分级分离来纯化类风湿因子，得到了一种高活性制剂，其由约95%的19Sγ球蛋白组成，未检测到7S污染物。通过对类风湿血清以及先前经阴离子交换层析分离的巨球蛋白进行阳离子交换层析（羧甲基纤维素），获得了存在两种因子的证据。一种因子（因子I）能凝集致敏绵羊细胞和乳胶颗粒，也能与人II组分沉淀。第二种因子（因子II）能凝集乳胶颗粒并与II组分沉淀，但不能凝集致敏绵羊细胞。通过阳离子交换层析在四份正常血清中的两份中证实了具有因子II特性的物质。有人提出，在某些异常血清中观察到的乳胶颗粒凝集及与人II组分沉淀可能是由于因子II浓度升高所致。
