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目的 为探讨血吸虫肠相关循环抗原CAA与CCA诊断靶微粒上表位特异性的差别 ,并试探获取其纯化制品用于定量检测的标准系列。方法 对两组分进行了亲和层析及阴离子交换剂高压液相纯化分离 ,并应用单抗检测系统进行同相和异相交互测试。结果 Mono QHPLC梯度洗脱分离AWAj-TCA可溶组分 ,可获得一个带阳离子活性的非结合CCA - 1峰及 3个大小不等的 ,带阳离子活性的CCA - 2 ,CCA - 3,CCA - 4非结合洗脱峰 ,以及一个带阴离子活性CAA - 1洗脱峰。CAA活性峰在峰谱上与CCA - 3有部分重叠。与单抗亲和层析纯品的活性对比测定显示CCA - 1与CCA - 2为该组分的主要构成 ,但CCA - 2及CAA - 1在本实验条件下均有微量的相互杂染。同相和异相双位点ELISA的 4种组合交互检测 ,展示CCA只能在捕获与检测抗体同为抗CCA单抗的一种组合中被检示 ,而另 3种组合都只能测出CAA组分。结论 血吸虫肠相关CCA组分为一兼含两性电荷的分子混合体 ;而CAA分子上具有一个可被抗CCA单抗识别的活性表位位点 ,从而可能影响纯化分离和特异检测。
Objective To investigate the specificity of the epitope specificity of CAA and CCA for the diagnosis of schistosoma intestinal-associated circulating antigen (CAA), and to explore the standard series of the purified products for quantitative detection. Methods The two components were purified by affinity chromatography and anion exchange liquid high pressure liquid phase separation, and the single phase antibody detection system was used to conduct in-phase and out-of-phase interaction tests. Results Mono QHPLC gradient elution was used to separate the AWAj-TCA soluble fraction. A non-binding CCA - 1 peak with cationic activity and three cationic active CCA - 2, CCA - 3 and CCA - 4 non-binding elution peak and one peak with anion-reactive CAA-1. The peak of CAA activity partially overlaps with CCA - 3 in the peak spectrum. The results of CCA - 1 and CCA - 2 assay showed that CCA - 2 and CAA - 1 were slightly mixed with each other in this experiment. Four combinations of in-phase and out-of-phase two-site ELISA were detected interactively, demonstrating that CCA can only be detected in a combination of capture and detection antibodies with anti-CCA mAb, while the other three combinations only detect CAA Component. Conclusions The schistosome enterohepatic CCA fraction is a mixture of amphipathic molecules. However, the CAA molecule has an active epitope that can be recognized by anti-CCA monoclonal antibody, which may affect the purification and isolation and specific detection.