Perego R, Vezzoli S, Manfredi W, Bigi G, Procaccia S
Boll Ist Sieroter Milan. 1983 Nov 30;62(5):462-8.
We studied soluble immune complexes in sera and synovial fluids of ten patients suffering from Rheumatoid Arthritis (RA), six seropositive and four seronegative. Isolation of the immune complexes was obtained by a combination of gel filtration and affinity chromatography from biological fluids. The first step involved preparation of a globulin-enriched fraction by precipitation of either serum or synovial fluid with 5% polyethylene glycol 6000. The precipitate was solubilized in Borate NaCl buffer and fractioned by gel filtration in Ultrogel AcA 34. Eluted peaks were submitted to affinity chromatography on a Protein A-Sepharose CL 4B column. The recovered material was analyzed by double immunodiffusion and electrophoresis in 6-20% polyacrylamide gel containing sodium dodecylsulphate (SDS-PAGE) and was shown to contain IgG, IgM, IgA with small amounts of C1q, C3 and C4. Observation occurred that there is a different distribution of macromolecular and intermediate complexes in seropositive and seronegative samples respectively, total C1q binding activity being equal.
我们研究了10例类风湿关节炎（RA）患者血清和滑液中的可溶性免疫复合物，其中6例血清阳性，4例血清阴性。通过凝胶过滤和亲和层析相结合的方法从生物体液中分离免疫复合物。第一步是用5%聚乙二醇6000沉淀血清或滑液制备富含球蛋白的部分。沉淀物溶解在硼酸盐氯化钠缓冲液中，并用Ultrogel AcA 34进行凝胶过滤分级分离。洗脱峰在蛋白A-琼脂糖CL 4B柱上进行亲和层析。回收的物质通过双向免疫扩散和在含十二烷基硫酸钠的6-20%聚丙烯酰胺凝胶中进行电泳（SDS-PAGE）分析，结果显示含有IgG、IgM、IgA以及少量的C1q、C3和C4。观察发现，血清阳性和血清阴性样本中大分子和中等复合物的分布分别不同，而总C1q结合活性相等。
