Janatova J, Gobel R J
J Immunol Methods. 1985 Dec 17;85(1):17-26. doi: 10.1016/0022-1759(85)90270-4.
The degree of the activation and fragmentation of C4 and C3, including chain structure of the activation products, was evaluated by SDS-PAGE analysis of the C4 or C3 antigens that were withdrawn from the reaction media with appropriate immunoadsorbent beads. Full activation of C4 and C3, and subsequent quantitative conversion of C4b into C4c, and C3b into iC3b took place in fresh NHS after the activation of complement with both aggIgG and CVF. For complete conversion of iC3b to C3c erythrocytes carrying the C3b receptor were added to the already activated serum. Both C4c and C3c were isolated by a 2-step procedure involving (i) an adsorption to and (ii) electrophoretic desorption from the respective immunoadsorbent beads.
通过用适当的免疫吸附珠从反应介质中提取C4或C3抗原进行SDS-PAGE分析，评估C4和C3的激活程度和片段化情况，包括激活产物的链结构。在用聚集免疫球蛋白G（aggIgG）和眼镜蛇毒因子（CVF）激活补体后，新鲜正常人血清（NHS）中发生了C4和C3的完全激活，随后C4b定量转化为C4c，C3b定量转化为iC3b。为了使iC3b完全转化为C3c，将携带C3b受体的红细胞加入到已经激活的血清中。C4c和C3c均通过两步法分离，第一步是吸附到各自的免疫吸附珠上，第二步是从免疫吸附珠上进行电泳解吸。
