Heyman B, Holmquist G, Borwell P, Heyman U
J Immunol Methods. 1984 Mar 30;68(1-2):193-204. doi: 10.1016/0022-1759(84)90150-9.
A simple ELISA assay for detecting murine anti-SRBC antibodies of IgG class was developed and the variation of the results according to different experimental conditions was investigated. Erythrocytes were left to settle in flexible plastic microtiter plates, after which they were fixed with glutaraldehyde and the remaining binding sites in the plates saturated with ovalbumin. Serum or monoclonal IgG antibodies were then allowed to react with the erythrocytes. Protein A coupled to alkaline phosphatase caused a color change in the subsequently added enzyme substrate. The results proved to be of good reproducibility, specificity and sensitivity. The assay can be used for measuring IgG concentration, estimating antibody avidity and number of antigenic determinants on the SRBC, as well as screening IgG anti-SRBC hybridomas. The precision of concentration estimates was very good when standard curves were used.
开发了一种用于检测IgG类小鼠抗SRBC抗体的简单ELISA检测方法，并研究了不同实验条件下结果的变化。将红细胞留在柔性塑料微量滴定板中沉降，之后用戊二醛固定，并用卵清蛋白饱和板中剩余的结合位点。然后使血清或单克隆IgG抗体与红细胞反应。与碱性磷酸酶偶联的蛋白A在随后添加的酶底物中引起颜色变化。结果证明具有良好的重复性、特异性和敏感性。该检测方法可用于测量IgG浓度、估计抗体亲和力和SRBC上抗原决定簇的数量，以及筛选IgG抗SRBC杂交瘤。使用标准曲线时，浓度估计的精度非常好。
