Groth T, Campbell E J, Herrmann K, Seifert B
School of Dentistry, Humboldt University, Berlin, Germany.
Biomaterials. 1995 Sep;16(13):1009-15. doi: 10.1016/0142-9612(95)94909-5.
In this study enzyme immunoassays are presented for the assessment of platelet adhesion/activation and fibrinogen adsorption/conformation. The estimation of platelet adhesion and activation was performed with two enzyme immunoassays (EIAs) using monoclonal antibodies directed against CD42b (GP lb) and CD 62 (GMP 140 or P-Selectin). The applicability of EIA was first demonstrated in microtitre plates coated with fibrinogen. The thrombogenic substrate showed that platelet adhesion and activation reached a plateau level within 30 min. The use of EIA for testing biomaterials was demonstrated with polymeric reference materials where a differentiation of materials with respect to adhesion and activation was achieved. To validate the EIA scanning electron microscopy was applied and confirmed the different extent of adhesion and activation of platelets on reference materials. In addition, polyurethaneureas, based on 4,4'-diphenylmethane diisocyanate and polytetramethylene glycols, with different hard segment content and composition were investigated. It was found that both adhesion and activation were not simply dependent on the hard segment content but also on the hard segment composition. To get more insight into the mechanism of this process, two EIAs for the binding of fibrinogen using polyclonal and monoclonal antibodies were developed. There it was shown that the adhesion and activation of platelets on polyurethaneureas was not simply dependent on the total amount of adsorbed fibrinogen but rather on its conformation, indicated by the binding of the monoclonal antibody directed vs the gamma-chain of fibrinogen.
在本研究中，介绍了用于评估血小板黏附/活化以及纤维蛋白原吸附/构象的酶免疫测定法。使用针对CD42b（糖蛋白Ib）和CD62（颗粒膜蛋白140或P-选择素）的单克隆抗体，通过两种酶免疫测定法（EIA）对血小板黏附和活化进行评估。EIA的适用性首先在包被有纤维蛋白原的微量滴定板中得到证实。血栓形成底物显示血小板黏附和活化在30分钟内达到稳定水平。通过聚合物参考材料证明了EIA在测试生物材料方面的应用，其中实现了材料在黏附和活化方面的区分。为了验证EIA，应用扫描电子显微镜并证实了血小板在参考材料上黏附和活化的不同程度。此外，研究了基于4,4'-二苯基甲烷二异氰酸酯和聚四亚甲基二醇、具有不同硬段含量和组成的聚氨酯脲。发现黏附和活化不仅简单地取决于硬段含量，还取决于硬段组成。为了更深入了解这一过程的机制，开发了两种使用多克隆和单克隆抗体检测纤维蛋白原结合的EIA。结果表明，血小板在聚氨酯脲上的黏附和活化不仅简单地取决于吸附纤维蛋白原的总量，还取决于其构象，这通过针对纤维蛋白原γ链的单克隆抗体的结合来表明。
