Barkas T, Al-Khateeb S F
Immunology. 1979 Feb;36(2):299-306.
An imporved assay system, using inhibition of ADCC, for the detection of 0.6 microgram/ml. of aggregated IgG in medium is described. Under these conditions, normal human serum is extremely inhibitory, this effect being attributed mainly to the weak binding of monomeric IgG. Enhancement of ADCC by albumin was also observed. Using this assay system without further modification, immune complexes may be detected in gel-fractionated sera by the distribution of the inhibitory material relative to the IgG peak. However, for routine use in the assay of untreated sera, a modification of the method is required. Preincubation of the spleen cells with the serum in the presence of EDTA, followed by a washing stage, reduces the background inhibition by normal sera to a low level. Effects of serum factors in producing variable amounts of background inhibition or stimulation are also avoided. The modified assay is sensitive to 6 microgram/ml of aggregated IgG in neat serum.
本文描述了一种改进的检测系统,该系统利用抗体依赖的细胞介导的细胞毒性(ADCC)抑制作用来检测培养基中0.6微克/毫升的聚集IgG。在这些条件下,正常人血清具有极强的抑制作用,这种作用主要归因于单体IgG的弱结合。还观察到白蛋白对ADCC有增强作用。使用该检测系统无需进一步改进,通过抑制物质相对于IgG峰的分布,可在凝胶分离的血清中检测免疫复合物。然而,对于未经处理血清检测的常规应用,需要对方法进行改进。在EDTA存在的情况下,将脾细胞与血清预孵育,随后进行洗涤步骤,可将正常血清的背景抑制降低到较低水平。还可避免血清因子产生不同程度背景抑制或刺激的影响。改进后的检测方法对纯血清中6微克/毫升的聚集IgG敏感。