Barclay Johanna L, Keshvari Sahar, Whitehead Jonathan P, Inder Warrick J
Mater Research Institute, University of Queensland, Brisbane, Australia Translational Research Institute, Brisbane, Australia
Mater Research Institute, University of Queensland, Brisbane, Australia Translational Research Institute, Brisbane, Australia School of Medicine, the University of Queensland, Brisbane, Australia.
Ann Clin Biochem. 2016 Sep;53(Pt 5):606-10. doi: 10.1177/0004563216628891. Epub 2016 Jan 8.
Thrombospondin-1 (TSP-1) is a circulating matricellular glycoprotein produced from many cell types including platelets. Currently TSP-1 is measured in either plasma or serum, using expensive commercial assays.
To develop and validate a cost effective in-house immunoassay for human TSP-1 suitable for quantitating levels from both plasma and serum.
An in-house enzyme-linked immunosorbent assay (ELISA) was developed for the measurement of human TSP-1. Sixteen healthy volunteers (8 male and 8 female), mean age 29 years (range 21-49), body mass index (BMI) mean 23.3 kg/m(2) (range 17.3-26.7) had non-fasted venous blood sampled at 0800 h and 1600 h for both plasma and serum TSP-1.
The assay limit of quantitation was 7.8 μg/L, inter assay CV was 17-31%, intra assay CV was 3-4% for plasma and <9% for serum. Plasma TSP-1 ranged from 133 to 478 μg/L (mean concentration 290 μg/L) in normal volunteers. Serum TSP-1 was approximately 100-fold higher, ranging from 13,700 to 44,400 μg/L (mean concentration 257,00 μg/L). There was no correlation between plasma and serum TSP-1.
TSP-1 can be readily measured in human plasma using ELISA. Serum concentrations are 100-fold higher, reflecting documented TSP-1 release by platelets, and does not provide a meaningful measure of circulating concentrations.
血小板反应蛋白-1（TSP-1）是一种由包括血小板在内的多种细胞类型产生的循环基质细胞糖蛋白。目前，TSP-1是使用昂贵的商业检测方法在血浆或血清中进行测量的。
开发并验证一种经济高效的人TSP-1内部免疫检测方法，适用于定量血浆和血清中的水平。
开发了一种用于测量人TSP-1的内部酶联免疫吸附测定（ELISA）。16名健康志愿者（8名男性和8名女性），平均年龄29岁（范围21-49岁），体重指数（BMI）平均为23.3kg/m²（范围17.3-26.7），于08:00和16:00采集非空腹静脉血，用于检测血浆和血清中的TSP-1。
该检测方法的定量下限为7.8μg/L，批间变异系数为17%-31%，血浆批内变异系数为3%-4%，血清批内变异系数<9%。正常志愿者血浆TSP-1范围为133至478μg/L（平均浓度290μg/L）。血清TSP-1约高100倍，范围为13700至44400μg/L（平均浓度25700μg/L）。血浆和血清TSP-1之间无相关性。
使用ELISA可轻松测量人血浆中的TSP-1。血清浓度高100倍反映了血小板释放TSP-1的记录情况，不能提供有意义的循环浓度测量值。
