Mazza G, Day M J, Barker R N, Corato A, Elson C J
University of Bristol, Department of Pathology and Microbiology, UK.
Autoimmunity. 1996;23(4):245-55. doi: 10.3109/08916939608995347.
A quantitative and sensitive cellular enzyme-linked immunosorbent assay was developed for determining the number of molecules of IgG of each subclass bound to the surface of murine red blood cells (RBC). To develop standard titration curves, RBC from normal mice were treated with tannic acid and coated with a known concentration of purified myeloma of each IgG subclass. The quantity of each subclass bound to the surface of erythrocytes was determined by calculating the protein concentration of the bound IgG, which was then converted into number of molecules of IgG/RBC. The assay was used to quantify the number of autoantibodies of all four IgG subclass bound to the erythrocytes of mice injected with rat RBC. Twenty one days after the first immunisation, a mean number of 84,000 molecules of IgG1/RBC were detected, which increased to 114,500 molecules/RBC on day 28. On days 56 and 96 the mean concentration of IgG1 remained high, however by day 110 the mean level of IgG1 had decreased slighty to 69,500 molecules/RBC. By contrast, the mean concentration of IgG2a autoantibodies was considerably lower throughout the experiment, starting at 40,200 molecules/RBC on day 21 and dropping to 2,500 molecules/RBC by day 110. The mean quantities of IgG2b and IgG3 autoantibodies were similar to each other, and intermediate between the levels of IgG1 and IgG2a autoantibodies.
开发了一种定量且灵敏的细胞酶联免疫吸附测定法，用于测定结合到小鼠红细胞（RBC）表面的各亚类IgG分子数量。为绘制标准滴定曲线，将正常小鼠的红细胞用单宁酸处理，并用已知浓度的各IgG亚类纯化骨髓瘤蛋白包被。通过计算结合的IgG的蛋白质浓度来确定结合到红细胞表面的各亚类的量，然后将其换算为IgG/红细胞的分子数。该测定法用于定量注射大鼠红细胞的小鼠红细胞上结合的所有四种IgG亚类自身抗体的数量。首次免疫后21天，检测到平均84,000个IgG1/红细胞分子，到第28天增加到114,500个分子/红细胞。在第56天和第96天，IgG1的平均浓度仍然很高，然而到第110天，IgG1的平均水平略有下降至69,500个分子/红细胞。相比之下，在整个实验过程中，IgG2a自身抗体的平均浓度要低得多，从第21天的40,200个分子/红细胞开始，到第110天降至2,500个分子/红细胞。IgG2b和IgG3自身抗体的平均量彼此相似，且介于IgG1和IgG2a自身抗体水平之间。
