Kofler R, Wick G
J Immunol Methods. 1977;16(3):201-9. doi: 10.1016/0022-1759(77)90198-3.
The influence of various methodological variables on the CrCl3 technique for coupling antigen to the surface of erythrocytes has been investigated. Tests should be performed in protein- and phosphate-free medium. The CrCl3 stock solution should be stored at a pH of 5.0 and working dilultions prepared in acetate buffer (pH 5.5). The coupling procedure itself was performed as described by Goding (1976) with slight modifications. The relation between antigen and CrCl3 concentrations was found to be of crucial importance as excess of antigen inactivates CrCl3 whereas lack of antigen or excess of CrCl3, leads to spontaneous agglutination. A preliminary test based on determination of the minimum concentration of CrCl3 which produces spontaneous agglutination in the absence of antigen, and coupling if antigen is available, can be employed to predict the optimal concentration with sheep, human or chicken red blood cells. The pH-dependency of the coupling process is emphasized.
已研究了各种方法学变量对用三氯化铬（CrCl3）技术将抗原偶联到红细胞表面的影响。测试应在无蛋白和无磷酸盐的培养基中进行。三氯化铬储备液应在pH 5.0下储存，并在乙酸盐缓冲液（pH 5.5）中制备工作稀释液。偶联程序本身按照戈丁（1976年）所述进行，略有修改。发现抗原与三氯化铬浓度之间的关系至关重要，因为过量的抗原会使三氯化铬失活，而抗原不足或三氯化铬过量会导致自发凝集。基于测定在无抗原情况下产生自发凝集的三氯化铬最低浓度，并在有抗原时进行偶联的初步试验，可用于预测绵羊、人或鸡红细胞的最佳浓度。强调了偶联过程的pH依赖性。
