ADAMTS13 content and VWF multimer and triplet structure in commercially available VWF/FVIII concentrates

Kannicht, Christoph; Fisseau, Claudine; Hofmann, Werner; Kröning, Mario; Fuchs, Birte

doi:10.1016/j.biologicals.2014.11.006

Cited by 5 publications

(5 citation statements)

References 38 publications

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“…This metalloproteinase regulates the multimeric size of VWF in plasma by cleaving the protein at a site in the A2 domain which lies between the GPIb and collagen binding sites in the A1 and A3 domains, respectively . It has been reported that Willfactin and Wilate contain relatively low levels of ADAMTS13 and have a VWF triplet structure similar to that of normal plasma . Additionally, this study also demonstrated that Fandhi and Haemate‐P had higher ADAMTS13 activity with an altered triplet structure.…”

Section: Discussionsupporting

confidence: 72%

“…28 It has been reported that Willfactin and Wilate contain relatively low levels of ADAMTS13 and have a VWF triplet structure similar to that of normal plasma. 29 Additionally, this study also demonstrated that Fandhi and Haemate-P had higher ADAMTS13 activity with an altered triplet structure. Alphanate is also reported to have high ADAMTS13 activity.…”

Section: Discussionmentioning

confidence: 53%

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Evaluation of von Willebrand factor concentrates by platelet adhesion to collagen using an in vitro flow assay

Riddell

Vinayagam

Gomez

et al. 2019

Research and Practice in Thrombosis and Haemostasis

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BackgroundVon Willebrand disease (VWD) results from quantitative or qualitative deficiency of von Willebrand factor (VWF) and is treated using VWF‐containing concentrates. Several studies have compared the function of various VWF containing concentrates however this has not been performed using shear based assays.ObjectivesTo compare the platelet‐capture potential of 10 commercially available, plasma‐derived VWF concentrates under shear conditions.Methods VWF containing concentrates were assessed for VWF:Ag, VWF:CB, VWF:RCo, factor VIII:C ADAMTS13 content, VWF multimeric profile and glycan content using lectin binding assays. Free‐thiol content of each concentrate was investigated using MPB binding assays. An in vitro flow assay was used to determine the ability of each concentrate to mediate platelet capture to collagen.Results VWF multimeric analysis revealed reduction of high molecular weight (HMW) forms in four of the concentrates (Alphante, Octanate and Haemoctin, and 8Y). The high MW multimer distribution of the remaining six concentrates (Optivate, Wilate, Fandhi, Wilfactin, Haemate P, and Voncento) was similar to the plasma control. Lectin analysis demonstrated that 8Y had increased amount of T‐antigen. Although platelet capture after 5 minutes perfusion was similar for all concentrates; Alphante, Octanate, and Haemoctin, demonstrated the lowest levels of platelet capture after 60 seconds of perfusion. Free‐thiol content and ADAMTS13 levels varied widely between the concentrates but was not correlated with function.ConclusionAlphanate, Octanate, and Haemoctin, lacked HMW multimers and had the lowest initial platelet capture levels suggesting that the presence of VWF HMW multimers are required for initial platelet deposition.

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Section: Discussionsupporting

confidence: 72%

Section: Discussionmentioning

confidence: 53%

Evaluation of von Willebrand factor concentrates by platelet adhesion to collagen using an in vitro flow assay

Riddell

Vinayagam

Gomez

et al. 2019

Research and Practice in Thrombosis and Haemostasis

View full text Add to dashboard Cite

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“…7 VWF protein is synthesised as a monomeric peptide that undergoes post-translational multimerisation in the golgi apparatus to tetramer, to hexamer and further to very-large multimer forms, 9 with overall molecular weight (MW) ranging from 800 to 20 000 kDa. 10 The regulatory metalloproteinase, ADAMTS13 (A Disintegrin And Metalloprotease with ThromboSpondin Type 1 motifs, member 13), cleaves large multimers into smaller less adhesive forms, generating characteristic low (LMWM), intermediate (IMWM) and high molecular weight multimer (HMWM) forms 11 (the largest being most functionally effective 9 ), as well as the triplet sets of protein bands demonstrated by high resolution gel chromatography assays. 10,12 Following tissue injury, VWF binds collagen at the subendothelium, where influences of blood flow shear stress expose glycoprotein Ib (GPIb) binding sites, thereafter attaching to and activating platelets, and initiating formation of the platelet plug.…”

Section: Introductionmentioning

confidence: 99%

Semi‐automated von Willebrand factor multimer assay for von Willebrand disease: Further validation, benefits and limitations

Oliver

Vanniasinkam

Mohammed

et al. 2019

Int J Lab Hematology

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Introduction: Accurate diagnosis of von Willebrand disease (VWD) enables effective patient management. von Willebrand factor (VWF) multimer analysis provides useful information regarding VWF multimer structure, thereby aiding VWD subtyping and management; however, historically technically challenging assays have had limited utility. This study evaluates the Sebia Hydrasys Hydragel-11 semi-automated VWF multimer assay and further validates the Hydragel-5 gel system, as primarily pertaining to VWD diagnostics and monitoring of therapy. Methods: Provisionally diagnosed (via a reference assay test panel) archived patientsamples and prospective test patient samples, including those undergoing desmopressin trial or therapy monitoring, along with commercial and in-house control material and various external quality assessment (EQA) samples, were analysed. VWF multimers were evaluated for presence, loss or partial loss of high molecular weight (HMWM) and intermediate molecular weight (IMWM) multimers by both visual inspection and densitometric scanning, and comparison with reference assay results. Results: All anticipated multimer patterns were reproduced, with patients generally showing multimer profiles matching expected patterns according to VWD type based on reference test panel 'diagnosis'. Occasional discrepancies were resolved by retesting. The increase in plasma VWF following desmopressin therapy was also clearly demonstrated. Multimer profiles of EQA samples complemented reference test panel results and matched EQA targets. There were some 'technical' limitations noted. Conclusion: This easy to use, standardised, semi-automated multimer analysis system can demonstrate the multimer profile of VWD patients, thus representing an additional laboratory tool for improved diagnosis, thereby facilitating appropriate patient management. K E Y W O R D S desmopressin, Hydragel, Multimers, von Willebrand disease, von Willebrand factor | 763 OLIVER Et aL.

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“…Plasma VWF multimers are modified by proteolytic cleavage with ADAMTS13, which generates N‐terminal and C‐terminal fragments 7,8,42,43 . These cleaved fragments are known as the faster migrating satellite, intermediate triplet, and slower migrating satellite based on their mobility in multimer analysis using SDS‐agarose gel electrophoresis 44,45 .…”

Section: Discussionmentioning

confidence: 99%

VWF‐Gly2752Ser, a novel non‐cysteine substitution variant in the CK domain, exhibits severe secretory impairment by hampering C‐terminal dimer formation

Okamoto

Tamura

Sanda

et al. 2022

Journal of Thrombosis and Haemostasis

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Background Von Willebrand factor (VWF) is a multimeric glycoprotein that plays important roles in hemostasis and thrombosis. C‐terminal interchain‐disulfide bonds in the cystine knot (CK) domain are essential for VWF dimerization. Previous studies have reported that missense variants of cysteine in the CK domain disrupt the intrachain‐disulfide bond and cause type 3 von Willebrand disease (VWD). However, type 3 VWD‐associated noncysteine substitution variants in the CK domain have not been reported. Objective To investigate the molecular mechanism of a novel non‐cysteine variant in the CK domain, VWF c.8254 G>A (p.Gly2752Ser), which was identified in a patient with type 3 VWD as homozygous. Methods Genetic analysis was performed by whole exome sequencing and Sanger sequencing. VWF multimer analysis was performed using SDS‐agarose electrophoresis. VWF production and subcellular localization were analyzed using ex vivo endothelial colony forming cells (ECFCs) and an in vitro recombinant VWF (rVWF) expression system. Results The patient was homozygous for VWF‐Gly2752Ser. Plasma VWF enzyme‐linked immunosorbent assay showed that the VWF antigen level of the patient was 1.2% compared with healthy subjects. A tiny amount of VWF was identified in the patient's ECFC. Multimer analysis revealed that the circulating VWF‐Gly2752Ser presented only low molecular weight multimers. Subcellular localization analysis of VWF‐Gly2752Ser‐transfected cell lines showed that rVWF‐Gly2752Ser was severely impaired in its ER‐to‐Golgi trafficking. Conclusion VWF‐Gly2752Ser causes severe secretory impairment because of its dimerization failure. This is the first report of a VWF variant with a noncysteine substitution in the CK domain that causes type 3 VWD.

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ADAMTS13 content and VWF multimer and triplet structure in commercially available VWF/FVIII concentrates

Cited by 5 publications

References 38 publications

Evaluation of von Willebrand factor concentrates by platelet adhesion to collagen using an in vitro flow assay

Evaluation of von Willebrand factor concentrates by platelet adhesion to collagen using an in vitro flow assay

Semi‐automated von Willebrand factor multimer assay for von Willebrand disease: Further validation, benefits and limitations

VWF‐Gly2752Ser, a novel non‐cysteine substitution variant in the CK domain, exhibits severe secretory impairment by hampering C‐terminal dimer formation

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