دورية أكاديمية
أعرض في EDS

Nanobody activator improves sensitivity of the von Willebrand factor activity assay to multimer size.

التفاصيل البيبلوغرافية
العنوان: Nanobody activator improves sensitivity of the von Willebrand factor activity assay to multimer size.
المؤلفون: Liang Q; Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Parker ET; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Dean G; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Karpen MS; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Wu Y; Department of Statistics, School of Statistics and Center of Statistical Research, Southwestern University of Finance and Economics, Chengdu, China., Wang X; Department of Laboratory Medicine, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai, China., Di Paola J; Department of Pediatrics, Washington University School of Medicine, St. Louis, Missouri, USA., Maier CL; Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, Georgia, USA., Meeks SL; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Lollar P; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Sidonio RF; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA., Li R; Aflac Cancer and Blood Disorders Center, Children's Healthcare of Atlanta, Department of Pediatrics, Emory University School of Medicine, Atlanta, Georgia, USA. Electronic address: renhao.li@emory.edu.
المصدر: Journal of thrombosis and haemostasis : JTH [J Thromb Haemost] 2024 Jul; Vol. 22 (7), pp. 2052-2058. Date of Electronic Publication: 2024 May 03.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Elsevier Country of Publication: England NLM ID: 101170508 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1538-7836 (Electronic) Linking ISSN: 15387836 NLM ISO Abbreviation: J Thromb Haemost Subsets: MEDLINE
أسماء مطبوعة: Publication: 2023- : [New York] : Elsevier
Original Publication: Oxford : Blackwell Pub.
مواضيع طبية MeSH: von Willebrand Factor*/metabolism , von Willebrand Factor*/analysis , Single-Domain Antibodies*/immunology , Single-Domain Antibodies*/chemistry , Enzyme-Linked Immunosorbent Assay* , Protein Multimerization*, Humans ; Linear Models ; Recombinant Proteins ; Blood Platelets/metabolism ; Platelet Aggregation/drug effects ; Platelet Glycoprotein GPIb-IX Complex/metabolism ; Protein Binding ; Platelet Adhesiveness ; Molecular Weight
مستخلص: Background: The activity of von Willebrand factor (VWF) in facilitating platelet adhesion and aggregation correlates with its multimer size. Traditional ristocetin-dependent functional assays lack sensitivity to multimer sizes. Recently, nanobodies targeting the autoinhibitory module and activating VWF were identified.
Objectives: To develop an assay that can differentiate the platelet-binding activity of VWF multimers.
Methods: A novel enzyme-linked immunosorbent assay (nanobody-triggered glycoprotein Ib binding assay [VWF:GPIbNab]) utilizing a VWF-activating nanobody was developed. Recombinant VWF, plasma-derived VWF (pdVWF), and selected gel-filtrated fractions of pdVWF were evaluated for VWF antigen and activity levels. A linear regression model was developed to estimate the specific activity of VWF multimers.
Results: Of the 3 activating nanobodies tested, 6C11 with the lowest activation effect exhibited the highest sensitivity for high-molecular-weight multimers (HMWMs) of VWF. VWF:GPIbNab utilizing 6C11 (VWF:GPIbNab 6C11 ) produced significantly higher activity/antigen ratios for recombinant VWF (>2.0) and HMWM-enriched pdVWF fractions (>2.0) than for pdVWF (∼1.0) or fractions enriched with shorter multimers (<1.0). The differences were much larger than those produced by VWF:GPIbNab utilizing other nanobodies, VWF:GPIbM, VWF:GPIbR, or VWF:CB assays. Linear regression analysis of 5 pdVWF fractions of various multimer sizes produced an estimated specific activity of 2.7 for HMWMs. The analysis attributed >90% of the VWF activity measured by VWF:GPIbNab 6C11 to that of HMWMs, which is significantly higher than all other activity assays tested.
Conclusion: The VWF:GPIbNab 6C11 assay exhibits higher sensitivity to HMWMs than ristocetin-based and collagen-binding assays. Future studies examining the application of this assay in clinical settings and any associated therapeutic benefit of doing so are warranted.
Competing Interests: Declaration of competing interests R.L. and Q.L. are coinventors on a pending patent describing applications of nanobodies to measure the von Willebrand factor activity. The other coauthors declare no conflicts of interest.
(Copyright © 2024 International Society on Thrombosis and Haemostasis. Published by Elsevier Inc. All rights reserved.)
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معلومات مُعتمدة: R35 HL166654 United States HL NHLBI NIH HHS
فهرسة مساهمة: Keywords: nanobody; platelet; ristocetin; sensitivity; von Willebrand factor
المشرفين على المادة: 0 (von Willebrand Factor)
0 (Single-Domain Antibodies)
0 (Recombinant Proteins)
0 (Platelet Glycoprotein GPIb-IX Complex)
تواريخ الأحداث: Date Created: 20240504 Date Completed: 20240630 Latest Revision: 20240702
رمز التحديث: 20240702
مُعرف محوري في PubMed: PMC11214880
DOI: 10.1016/j.jtha.2024.04.015
PMID: 38704122
قاعدة البيانات: MEDLINE
الوصف
تدمد:1538-7836
DOI:10.1016/j.jtha.2024.04.015