The online version of this article (doi:10.1186/s13045-017-0407-1) contains supplementary material, which is available to authorized users.
We developed a novel murine monoclonal antibody (mAb) against the C-terminal α-helix of the human von Willebrand factor A2, designated SZ-179. We showed that SZ-179 inhibited the interactions between VWF and ADAMTS13 and prevented the degradation of high molecular weight VWF multimers. Importantly, SZ-179 reduced the proteolysis of VWF-R1597W mutant by rADAMTS13 dose-dependently under native conditions. Our findings reveal a potential therapeutic target for bleeding disorders.
Additional file 1: Supplemental data. Detailed methods and materials are shown. (DOC 115 kb)13045_2017_407_MOESM1_ESM.doc
Additional file 2: Figure S1. Characterization of mAb SZ-179. (A) Quantification of ELISA analyses detecting SZ-179 binding to IgG1, IgG2a, IgG2a, IgG3, or IgM. (B) Quantification of ELISA analyses for SZ-179 or murine IgG1 binding to VWFα5. Dose–response curves are shown. (C) Quantification of ELISA analyses for SZ-179 or murine IgG1 binding to plasma-derived VWF. Dose–response curves are shown. Data are mean ± SD of four independent experiments. (DOCX 974 kb)13045_2017_407_MOESM2_ESM.docx
Additional file 3: Figure S2. SZ-179 inhibits cleavage of VWF by ADAMTS13 in plasma under denaturing conditions. (A, B) Pooled normal human plasma was pre-incubated with SZ-179 or isotype control IgG1 for 2 h at 37°C, and then incubated with 1.5M urea for 16 h. The proteolytic products were separated by electrophoresis in a 1.3% agarose gel and detected by anti-VWF. (C) Dose–response curve for inhibition of plasma ADAMTS13-mediated cleavage of plasma-VWF. (D) Dose–response curve for inhibition of rADAMTS13-mediated GST-VWF73-H cleavage. Results represented as mean ± SD of four independent experiments. (DOCX 1519 kb)13045_2017_407_MOESM3_ESM.docx
- A novel monoclonal antibody against the von Willebrand Factor A2 domain reduces its cleavage by ADAMTS13
- BioMed Central
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