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Development of a novel flow cytometric immunobead array to quantify VWF:Ag and VWF:GPIbR and its application in acute myocardial infarction.
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Development of a novel flow cytometric immunobead array to quantify VWF:Ag and VWF:GPIbR and its application in acute myocardial infarction.

Eur J Haematol. 2017 May 18;:

Authors: Yan B, Xu M, Zhao Y, Guo H, Xia L, Ruan C, Zhao Y

Abstract
OBJECTIVES: Both von Willebrand disease (VWD) and acute myocardial infarction (AMI) involve quantitative and qualitative changes in von Willebrand factor (VWF). Our objective was to develop a rapid and precise flow cytometric immunobead array (FCIA) to quantify VWF antigen (VWF:Ag) and ristocetin-triggered platelet glycoprotein Ib binding (VWF:GPIbR), and apply it in a clinical setting.
METHODS: Microbeads, coated with monoclonal antibodies for SZ29 or SZ151 IgG, were incubated with diluted plasma. VWF-binding microbeads were detected with FITC-conjugated sheep anti-human VWF IgG by flow cytometry. Plasma VWF:Ag and VWF:GPIbR levels in normal controls (CTL; n = 105), patients with VWD (n = 21), and patients with AMI (n = 146) were tested by FCIA and ELISA in parallel. ADAMTS13 activity and VWF multimer analyses were also implemented.
RESULTS: Our novel FCIA showed a strong correlation with the ELISA results (VWF:Ag, r = 0.855; VWF:GPIbR, r = 0.813). The intra-assay coefficient variations (CVs) of VWF:Ag-FCIA and VWF:GPIbR-FCIA were 9.2% and 7.7%, respectively, and the inter-assay CVs were 12.6% and 13.5%, respectively. Plasma VWF:Ag, and VWF:GPIbR levels were significantly higher in patients with AMI than in CTL (p < 0.0001), whereas the ratios of ADAMTS13/VWF:Ag and ADAMTS13/VWF:GPIbR were significantly lower (p < 0.0001). Levels of plasma ultra-large VWF (UL-VWF) were dramatically increased in patients with AMI.
CONCLUSIONS: The novel VWF:Ag and VWF:GPIbR FCIA assays were found to be simpler, more specific, and more accurate than the classical ELISA method. In addition, elevated VWF:GPIbR and UL-VWF may contribute to the pathogenesis of AMI. This article is protected by copyright. All rights reserved.

PMID: 28523822 [PubMed - as supplied by publisher]