NANOBODY TARGETING VON WILEBRAND FACTOR A3-DOMAIN

"Inventors have isolated a single-domain antibody (designated KB-VWF-D3.1) targeting the A3-domain, the epitope of which overlaps the collagen-binding site. Binding of KB-VWF-D3.1 proved independent of VWF multimer size. However, its interaction with VWF was lost upon proteolysis by ADAMTS13, suggesting that proteolysis in the A2-domain modulates exposure of its epitope in the A3-domain. Inventors therefore used KB-VWF-D3.1 to monitor VWF degradation in plasma samples. Spiking experiments showed that a loss of 10% intact-VWF could be detected using this single-domain antibody. By comparing plasma from volunteers to that of congenital VWD-patients, intact-VWF levels were significantly reduced for all VWD-types, and most severely in VWD-type 2A(IIA) in which mutations promote ADAMTS13- mediated proteolysis. Unexpectedly, low-grade proteolysis in VWD-type 1 and -type 2M was also observed. Thus, this single-domain antibody proved sensitive to detect low-grade degradation in plasma from patients with AVWS and congenital VWD, including types 1 and 2M.
The present invention relates to an isolated single domain antibody targeting at least one region of A3-domain of VWF, wherein the region comprising the following sequences: SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 and/or SEQ ID NO: 4. "

Keywords: VWF A3-domain, Nanobody, Diagnosis, von Willebrand's disease, ELISA
Patent Application number: European Procedure (Patents) (EPA) - 15 Déc. 2022 - 22 306 896.6 and PCT/EP2023/085828 on 14/12/2023
Inventors:
LENTING Petrus,SUSEN Sophie,CHRISTOPHE Olivier,DENIS Cécile,CASARI Caterina
Publications:
Blood, 2023 Mar 23;141(12):1457-1468., Kizlik-Masson et al, A nanobody against the VWF A3 domain detects ADAMTS13-induced proteolysis in congenital and acquired VWD doi: 10.1182/blood.2022017569.

Reference:

BIO22211-D1

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Patent filling date: 2022-12-15

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