Smith Brendon W, Strakova Jana, King Jennifer L, Erdman John W, O'Brien William D
Division of Nutritional Sciences.
Biomark Insights. 2010 Nov 28;5:119-27. doi: 10.4137/BMI.S6051.
von Willebrand Factor (vWF) is a multimeric plasma protein important for platelet plug formation. As part of its haemostatic role, it is released from endothelial cells during vascular stress or injury and is considered an excellent biomarker of endothelial function. Currently, there are no validated kits available to measure vWF in rabbits. We developed a sensitive and reproducible sandwich enzyme-linked immunosorbent assay (ELISA) for detection of vWF in rabbit plasma using commercially available antibodies and reagents. Purified human vWF was used as a calibrator standard with a dynamic range of 1.56-100 ng/mL. The Minimum Required Dilution for rabbit plasma was 1:100. When plasma was spiked with 3.76 or 10 ng/mL vWF, recovery was 108 ± 2% and 93 ± 2%, respectively. Intra- and inter-assay precision for 8 rabbit plasma samples were 3% and 4%, respectively. The Minimum Detectable Concentration was 254 pg/mL for purified human vWF and 1:10,700 dilution of cholesterol-fed rabbit plasma, and the Reliable Detection Limits were 457 pg/mL and 1:5940. Three freeze-thaw cycles significantly decreased vWF concentrations for purified human vWF and 2 of 3 plasma samples assayed. This ELISA provides sensitive and reproducible measurements of rabbit plasma vWF, which is an important biomarker for cardiovascular research.
血管性血友病因子（vWF）是一种对血小板栓子形成很重要的多聚体血浆蛋白。作为其止血作用的一部分，它在血管应激或损伤时从内皮细胞释放出来，被认为是内皮功能的优秀生物标志物。目前，尚无经过验证的试剂盒可用于检测兔体内的vWF。我们使用市售抗体和试剂开发了一种灵敏且可重复的夹心酶联免疫吸附测定（ELISA）法，用于检测兔血浆中的vWF。纯化的人vWF用作校准标准品，动态范围为1.56 - 100 ng/mL。兔血浆的最低所需稀释度为1:100。当血浆中加入3.76或10 ng/mL的vWF时，回收率分别为108±2%和93±2%。8个兔血浆样品的批内和批间精密度分别为3%和4%。纯化的人vWF和胆固醇喂养兔血浆1:10,700稀释液的最低可检测浓度为254 pg/mL，可靠检测限为457 pg/mL和1:5940。三个冻融循环显著降低了纯化的人vWF和所检测的3个血浆样品中的2个的vWF浓度。这种ELISA法能灵敏且可重复地检测兔血浆中的vWF，vWF是心血管研究中的一个重要生物标志物。
