Ziola B, Salmi A, Penttinen K
J Immunol Methods. 1982 Mar 26;49(3):247-60. doi: 10.1016/0022-1759(82)90125-9.
Experiments were undertaken to determine if porcine C1q could replace human C1q in the solid-phase immunoassay of human immune complexes (ICs). Porcine C1q was obtained by a two-cycle precipitation method involving dialysis against chelating agents in low ionic strength buffer. C1q was adsorbed to polystyrene beads and in vivo- or in vitro-formed ICs binding to the solid-phase C1q were detected with 125I-labeled or horseradish peroxidase-conjugated anti-human gamma antibodies. Unfractionated, heat-aggregated human gamma globulin (delta IgG) could be detected at 20 ng/ml when diluted in buffer only. The detection threshold changed to 40-80 ng delta IgG/ml when the assay was run with buffer containing normal human serum diluted 1: 1000 (the serum dilution used for detecting natural ICs). Analysis of systemic lupus erythematosus sera revealed that 60% contained highly significant levels of ICs (binding greater than or equal to 3 S.D. above the mean of controls). Comparison with platelet aggregation test results revealed a highly significant correlation between the two methods (P less than 0.0001), even though each assay detected ICs in several serum specimens negative in the other test. These results demonstrate that porcine C1q can functionally replace human C1q in the solid-phase immunoassay of human ICs. Since porcine blood is normally a waste product of the meat-processing industry, it is an obvious source of easily isolated C1q for use in such an assay.
开展实验以确定在人免疫复合物(ICs)的固相免疫测定中,猪C1q是否能够替代人C1q。猪C1q通过两步沉淀法获得,该方法包括在低离子强度缓冲液中用螯合剂进行透析。将C1q吸附到聚苯乙烯珠上,并用125I标记或辣根过氧化物酶偶联的抗人γ抗体检测与固相C1q结合的体内或体外形成的ICs。仅在缓冲液中稀释时,未分级的热聚集人γ球蛋白(δIgG)在20 ng/ml时即可被检测到。当用含有1:1000稀释的正常人血清的缓冲液进行检测(用于检测天然ICs的血清稀释度)时,检测阈值变为40 - 80 ng δIgG/ml。对系统性红斑狼疮血清的分析显示,60%含有高度显著水平的ICs(结合量比对照平均值高3个标准差以上)。与血小板聚集试验结果的比较显示,两种方法之间存在高度显著的相关性(P小于0.0001),尽管每种检测方法在其他检测为阴性的几个血清标本中都检测到了ICs。这些结果表明,在人ICs的固相免疫测定中,猪C1q在功能上可以替代人C1q。由于猪血通常是肉类加工业的废品,它显然是用于此类检测的易于分离的C1q的来源。