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Analytical characterization and reference interval of an enzyme-linked immunosorbent assay for active von Willebrand factor
- Source :
- PLoS ONE, Vol 14, Iss 2, p e0211961 (2019), PLoS ONE, PLOS ONE, 14(2):0211961. Public Library of Science
- Publication Year :
- 2019
- Publisher :
- Public Library of Science (PLoS), 2019.
-
Abstract
- BackgroundInteraction of von Willebrand factor (VWF) with platelets requires a conformational change that exposes an epitope within the VWF Al domain, enabling platelet glycoprotein Iba binding. Quantification of this "active" conformation of VWF has been shown to provide pathophysiological insight into conditions characterized by excessive VWF-platelet interaction.MethodsWe developed an immunosorbent assay based on a variable heavy chain antibody fragment against the VWF Al domain as a capture antibody. Assay performance in terms of specificity (binding to active R1306W- and sheared VWF), precision, accuracy, linearity, limits of detection and stability were determined. Active VWF, VWF antigen, VWF ristocetin cofactor activity, VWF:GP1bM and VWF propeptide were measured in citrated plasma and platelet-VWF binding in whole blood from 120 healthy individuals to establish a reference interval for active VWF and to assess associations with other VWF parameters.ResultsIntra- and inter-assay CVs were between 2.4-7.2% and 4.1-9.4%, depending on the level. Mean recovery of spiked recombinant R1306W VWF was 103 +/- 3%. The assay was linear in the range of 90.1-424.5% and had a limit of quantification of 101%. The reference interval for active VWF was 91.6-154.8% of NPP. Significant, positive correlations between active VWF and all other VWF parameters were found, with the strongest correlation with VWF: GP1bM binding.ConclusionsWe developed and validated an immunosorbent assay for the accurate detection of active VWF levels in plasma. The assay fulfilled all analytical criteria in this study and a reference interval was established, allowing its use to quantify active VWF in pathological conditions for future research.
- Subjects :
- VONWILLEBRAND-FACTOR
Male
Physiology
030204 cardiovascular system & hematology
Platelet membrane glycoprotein
DISEASE
Epitope
law.invention
Binding Analysis
Epitopes
0302 clinical medicine
law
Animal Cells
Reference Values
hemic and lymphatic diseases
BLOOD-GROUP
Blood plasma
Medicine and Health Sciences
Platelet
Enzyme-Linked Immunoassays
Shear Stresses
Whole blood
Multidisciplinary
biology
FACTOR-VIII
Chemistry
Physics
Classical Mechanics
ASSOCIATION
Middle Aged
ADAMTS13
Body Fluids
PLATELET-ADHESION
von Willebrand Diseases
Blood
Physical Sciences
Recombinant DNA
cardiovascular system
Mechanical Stress
Medicine
Female
Anatomy
Cellular Types
Research Article
circulatory and respiratory physiology
Platelets
Adult
Blood Platelets
congenital, hereditary, and neonatal diseases and abnormalities
Adolescent
Science
Enzyme-Linked Immunosorbent Assay
DIAGNOSIS
Research and Analysis Methods
Blood Plasma
Antibodies
03 medical and health sciences
Young Adult
AGE
Von Willebrand factor
FACTOR LEVEL
von Willebrand Factor
Humans
Protein precursor
Immunoassays
Chemical Characterization
Aged
Blood Cells
Biology and Life Sciences
Cell Biology
Molecular biology
Blood Counts
biology.protein
Immunologic Techniques
Blood Groups
030215 immunology
Subjects
Details
- Language :
- English
- ISSN :
- 19326203
- Volume :
- 14
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- PLoS ONE
- Accession number :
- edsair.doi.dedup.....ca77719612fcb1ac9c3734a98cb9254c