Moran N, Kiernan A, Dunne E, Edwards R J, Shields D C, Kenny D
Department of Clinical Pharmacology, Royal College of Surgeons in Ireland, Dublin 2, Ireland.
Anal Biochem. 2006 Oct 1;357(1):77-84. doi: 10.1016/j.ab.2006.06.037. Epub 2006 Jul 14.
Platelets play a central role in maintaining biological hemostasis. Inappropriate platelet activation is responsible for thrombotic diseases such as myocardial infarction and stroke. Therefore, novel agents that can inhibit platelet activation are necessary. However, assays that monitor platelet aggregation are generally time-consuming and require high volumes of blood and specialized equipment. Therefore, a medium- to high-throughput assay that can monitor platelet aggregation would be considered useful. Such an assay should be sensitive, comparable to the "gold standard" assay of platelet aggregometry, and able to monitor multiple samples simultaneously but with low assay volumes. We have developed such a microtiter assay. It can assay an average of 60 independent treatments per 60 ml blood donation and demonstrates greater sensitivity than the current gold standard assay, namely platelet aggregation in stirring conditions in a platelet aggregometer. The microtiter plate (MTP) assay can detect known inhibitors of platelet function such as indomethacin, aspirin, and ReoPro. It is highly reproducible when using standard doses of agonists such as thrombin receptor-activating peptide (20 microM) and collagen (0.19 mg/ml). Finally, the MTP assay is rapid and sensitive and can detect unknown platelet-modulating agents from a library of compounds.
血小板在维持生物止血过程中起着核心作用。不适当的血小板激活会导致诸如心肌梗死和中风等血栓性疾病。因此，需要能够抑制血小板激活的新型药物。然而，监测血小板聚集的检测方法通常耗时，需要大量血液和专门设备。因此，一种能够监测血小板聚集的中高通量检测方法将被认为是有用的。这样的检测方法应该灵敏，与血小板聚集测定的“金标准”检测方法相当，并且能够同时监测多个样本，但检测体积要小。我们已经开发了这样一种微量滴定检测方法。每60毫升献血平均可以检测60个独立的处理样本，并且比当前的金标准检测方法（即在血小板聚集仪中搅拌条件下的血小板聚集）具有更高的灵敏度。微量滴定板（MTP）检测方法可以检测已知的血小板功能抑制剂，如吲哚美辛、阿司匹林和ReoPro。当使用标准剂量的激动剂如凝血酶受体激活肽（20微摩尔）和胶原蛋白（0.19毫克/毫升）时，该方法具有高度的可重复性。最后，MTP检测方法快速且灵敏，能够从化合物库中检测未知的血小板调节剂。
