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Eur J Haematol. 2017 Sep;99(3):207-215. doi: 10.1111/ejh.12905. Epub 2017 Jun 21.

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

Yan B1,2, Xu M1,2, Zhao Y1,2, Guo H3, Xia L1,2,4, Ruan C1,2, Zhao Y1,2.

Author information

1
Jiangsu Institute of Hematology, Key Laboratory of Thrombosis and Hemostasis of Ministry of Health, The First Affiliated Hospital of Soochow University, Suzhou, China.
2
Collaborative Innovation Center of Hematology, Soochow University, Suzhou, China.
3
Department of Clinical Laboratory, The First Affiliated Hospital and College of Clinical Medicine, Henan University of Science and Technology, Luoyang, China.
4
Cardiovascular Biology Research Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA.

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=.855; VWF:GPIbR, r=.813). The intra-assay coefficient variations (CVs) of VWF:Ag-FCIA and VWF:GPIbR-FCIA were 9.2% and 7.7%, respectively, and the interassay 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<.0001), whereas the ratios of ADAMTS13/VWF:Ag and ADAMTS13/VWF:GPIbR were significantly lower (P<.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.

KEYWORDS:

acute myocardial infarction; flow cytometry; multimer analysis; von Willebrand disease; von Willebrand factor activity

PMID:
28523822
DOI:
10.1111/ejh.12905
[Indexed for MEDLINE]

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