Arvidsson S, Askendal A, Lindahl T L, Tengvall P
Division of Applied Physics, Department of Physics, Chemistry and Biology, Linköping University, SE-581 83 Linköping, Sweden.
Colloids Surf B Biointerfaces. 2008 Apr 1;62(2):214-9. doi: 10.1016/j.colsurfb.2007.10.003. Epub 2007 Oct 6.
Surface mediated immune complement activation can be detected by a variety of antibody utilizing methods such as ELISA, fluorescence- or radiolabelling techniques, QCM, and ellipsometry. In the present work we investigated how the common anticoagulants heparin, dalteparin, fondaparinux and sodium citrate affected the binding of anti-complement factor 3c (anti-C3c) on a model complement activator surface, immobilised IgG, after incubation in human blood serum. The results show, as expected, that different anticoagulants affect the antibody binding differently. Increasing amounts of heparin, dalteparin and sodium citrate in normal serum resulted in a decreasing anti-C3c binding. The antibody deposition was not sensitive for the fondaparinux concentration. Surprisingly high concentrations of anti-coagulantia were needed to completely eradicate the antibody binding. Experiments in EGTA-serum showed that anticoagulants interfered directly with both the classical and alternative pathways. Control C3a-des arg ELISA measurements show that the lowered antibody surface binding was not a result of complement depletion in serum. Kallikrein generation by hydrophilic glass surfaces was not affected by high anticoagulant concentrations.
表面介导的免疫补体激活可以通过多种利用抗体的方法检测，如酶联免疫吸附测定（ELISA）、荧光或放射性标记技术、石英晶体微天平（QCM）和椭偏仪。在本研究中，我们调查了常见抗凝剂肝素、达肝素、磺达肝癸钠和柠檬酸钠在人血清中孵育后，如何影响抗补体因子3c（抗C3c）在模型补体激活剂表面（固定化IgG）上的结合。结果表明，正如预期的那样，不同的抗凝剂对抗体结合的影响不同。正常血清中肝素、达肝素和柠檬酸钠的量增加导致抗C3c结合减少。抗体沉积对磺达肝癸钠浓度不敏感。令人惊讶的是，需要非常高浓度的抗凝剂才能完全消除抗体结合。在乙二胺四乙酸（EGTA）血清中的实验表明，抗凝剂直接干扰经典途径和替代途径。对照C3a-去精氨酸ELISA测量表明，抗体表面结合降低不是血清中补体耗竭的结果。亲水性玻璃表面激肽释放酶的生成不受高抗凝剂浓度的影响。
