APPLICATION OF ENZYME IMMUNOASSAYS FOR TESTING HEMOCOMPATIBILITY OF BIOMEDICAL POLYMERS
APPLICATION OF ENZYME IMMUNOASSAYS FOR TESTING HEMOCOMPATIBILITY OF BIOMEDICAL POLYMERS
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DOI：
10.1016/0142-9612(95)94909-5
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发表时间：
1995-09-01
期刊：
影响因子：
14
通讯作者：
SEIFERT, B
中科院分区：
文献类型：
--
作者：
GROTH, T;CAMPBELL, EJ;SEIFERT, B
在这项研究中，酶免疫分析法用于评估血小板粘附/活化和纤维蛋白原吸附/构象。使用针对CD42b （GP - Ib）和cd62 （GMP - 140或p -选择素）的单克隆抗体进行两种酶免疫测定（EIAs）来评估血小板粘附和活化。EIA的适用性首先在包被纤维蛋白原的微滴板上得到证实。血栓形成底物显示血小板粘附和活化在30分钟内达到平台水平。使用EIA来测试生物材料是用聚合物标准材料来证明的，在这种材料中，材料的粘附性和活化性得到了区分。为了验证，应用了EIA扫描电镜，证实了血小板在对照物质上不同程度的粘附和活化。此外，还研究了以4,4'-二苯基甲烷二异氰酸酯和聚四甲基二醇为基料的不同硬段含量和组成的聚氨酯聚氨酯。结果表明，粘附和活化不仅与硬段含量有关，还与硬段组成有关。为了更深入地了解这一过程的机制，我们开发了两种用于纤维蛋白原结合的多克隆抗体和单克隆抗体。研究表明，血小板在聚氨酯脲上的粘附和活化不仅仅取决于吸附纤维蛋白原的总量，而是取决于其构象，这是由单克隆抗体与纤维蛋白原γ链的结合所表明的。
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 Ib) 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.