Abstract Number: OC 51.4
Meeting: ISTH 2021 Congress
Background: Von Willebrand factor antigen (VWF:Ag) and functionality are key parameters in the diagnosis, subclassification and management of von Willebrand disease (VWD). Measurement of VWF:Ag and function (e.g. binding to collagen and platelet glycoprotein Ibα) requires different assays or platforms, which often take time and specific expertise to perform. Fiber optic – surface plasmon resonance (FO-SPR) may offer an easy, simple-to-use and fast technique to measure VWF parameters.
Aims: To develop an FO-SPR based measurement of VWF:Ag.
Methods: Gold-coated optical fibers containing a carboxyl self-assembling monolayer were functionalized through binding of our in-house monoclonal anti-VWF antibody 6D1. After blocking, a simple dip of the probe into the plasma sample allowed to capture VWF. Subsequent FO-SPR signal generation was performed using a biotinylated polyclonal anti-VWF antibody and signal amplification was done using goat anti-biotin coated gold nanoparticles. After full optimization of the assay, a calibration curve was constructed using normal human plasma. Next, using a VWD type 3 patient plasma sample, the lower limit of detection (LLOD) and quantification (LLOQ) were established. Finally, VWF:Ag levels of 11 VWD samples (all types included) were determined using the FO-SPR and compared to VWF:Ag levels measured in our in-house ELISA.
Results: All tested conditions revealed acceptable CV% values (<20%) and high signal to noise ratios (>10). LLOD and LLOQ were 0.68 and 1.24 µg/mL respectively. ELISA and FO-SPR-based measurements of the VWD samples (range: 1.38 – 7.46 µg/mL) showed a good correlation (p = 0.5771, r = 0.9014) with no systematic bias (mean bias: 2.417, 95%CI[-13.06, 17.89]).
Conclusions: A robust VWF:Ag FO-SPR-based assay was developed which showed a good correlation compared to our in-house ELISA. Currently, VWF:CB and VWF:RCo FO-SPR-based assays are being developed, which will ultimately allow a simultaneous fast (within 15-20min) and easy determination of the most important VWF parameters.
To cite this abstract in AMA style:Calcoen B, Dekimpe C, Jacquemin M, Tersteeg C, Vanhoorelbeke K, De Meyer S. Measurement of von Willebrand Factor Levels via Fiber Optic-surface Plasmon Resonance [abstract]. Res Pract Thromb Haemost. 2021; 5 (Suppl 2). https://abstracts.isth.org/abstract/measurement-of-von-willebrand-factor-levels-via-fiber-optic-surface-plasmon-resonance/. Accessed June 25, 2022.
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