Kao K J, Klein P A
Am J Clin Pathol. 1986 Sep;86(3):317-23. doi: 10.1093/ajcp/86.3.317.
An enzyme-linked immunosorbent assay was developed to quantitate the platelet secretory glycoprotein, thrombospondin (TSP), in plasma. Specificity of the assay was provided by using an anti-TSP monoclonal antibody (McAb) to coat 96 well polystyrene assay plates. The effective range of the standard curve for the assay was between 2.5 ng/mL and 500 ng/mL. Interassay and intraassay variations were 6.9% and 7.5%, respectively. To avoid spontaneous release of TSP from platelets after the blood was drawn, and to obtain a valid measurement of plasma TSP concentration, it was essential to place and maintain blood samples on ice immediately after blood was drawn and to separate plasma from cellular constituents within two hours. The mean plasma TSP concentration of 24 healthy adults was 64.3 +/- 18.0 ng/mL (mean +/- SD). The TSP concentration of pooled normal human serum (n = 6) was 15.9 micrograms/mL. Interference occurred when undiluted plasma or serum samples were assayed. This interfering effect could be eliminated completely by a tenfold dilution of plasma or serum samples before the assay. The assay also was applied successfully to quantitate TSP release from platelets after induction of aggregation by different platelet-aggregating agents.
开发了一种酶联免疫吸附测定法来定量血浆中的血小板分泌糖蛋白血小板反应蛋白（TSP）。该测定法的特异性通过使用抗TSP单克隆抗体（McAb）包被96孔聚苯乙烯测定板来实现。该测定法标准曲线的有效范围在2.5 ng/mL至500 ng/mL之间。批间和批内变异分别为6.9%和7.5%。为避免采血后血小板自发释放TSP，并获得血浆TSP浓度的有效测量值，采血后必须立即将血样置于冰上并保持，且在两小时内将血浆与细胞成分分离。24名健康成年人的血浆TSP平均浓度为64.3±18.0 ng/mL（平均值±标准差）。混合正常人血清（n = 6）的TSP浓度为15.9微克/毫升。测定未稀释的血浆或血清样品时会出现干扰。在测定前将血浆或血清样品进行十倍稀释可完全消除这种干扰作用。该测定法还成功应用于定量不同血小板聚集剂诱导聚集后血小板释放的TSP。
