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Blood, Vol. 106, Issue 3, 922-924, August 1, 2005
Localization of ADAMTS13 to the stellate cells of human liver
Blood Uemura et al.
106: 922
Supplemental materials for: Uemura et al, Vol 106, Issue 3, 922-924
Files in this Data Supplement:
- Figure S1. Characterization of two distinct anti-ADAMTS13 murine monoclonal antidodies (A10 and C7) (JPG, 45 KB)
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The immunoglobulin subclasses of A10 and C7 were IgG2b-κ and IgG1-κ, respectively. Full-length wild-type (WT) recombinant (r) ADAMTS13 and its 9 mutants that were truncated at their C-termini were produced as described previously. (A) Western blot analysis was performed using these rADAMTS13 proteins and the monoclonal antibodies of anti-Flag, A10, and C7 (UT indicates untransfected). Panel B shows that the epitopes recognized by the A10 and C7 antibodies were determined to reside in the disintegrin-like (D) domain and in the seventh and eighth thrombospondin type-1 (T7/T8) domains. Note that the neutralizing IgG autoantibodies of patients with acquired idiopathic thrombotic thrombocytopenic purpura commonly bind to the cys-rich/spacer (C and Sp) domains, as previously shown (panel B). (C) During inhibition assays, residual ADAMTS13 activity was measured after normal pooled plasma was incubated with various amounts of the purified monoclonal IgGs for 2 hours at 37°C. The A10 antibodies inhibited plasma ADAMTS13 activity in a dose-dependent manner, with complete inhibition observed at IgG 50 µg/mL (final). The C7 antibodies only partially inhibited ADAMTS13 activity even when the final concentration of the monoclonal antibodies was raised to IgG 100 µg/mL.
1Soejima K, Matsumoto M, Kokame K, et al. ADAMTS13 cystein-rich/spacer domains are functionally essential for von Willebrand factor cleavage. Blood. 2003;102-3232-3237.
2Ibid.
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