Hi-Puri™ Anti-Human β-tryptase monoclonal antibody

(A) Binding kinetics of anti-tryptase clone 31A.v11 to human tryptases.
(B and C) Tryptase activity using peptide substrate S-2288 for βI-, βII-, βIII-tryptases with anti-tryptase clone 31A.v11 (B) and for βI-tryptase with anti-tryptase clone 31A.v11 Fab or IgG4 antibody (C).
(D) Tryptase activity using ES002 peptide substrate with degranulated foreskin MC releasate with anti-tryptase clone 31A.v11 Fab or IgG4.

(H) Primary human SMC proliferation stimulated by βI-tryptase with or without anti-tryptase.
(I) Primary human SMC contraction induced by βI-tryptase with or without anti-tryptase.
(J) Collagen secretion by primary human lung fibroblasts induced by recombinant βI-tryptase treated with anti-tryptase or medium.

The activity-based probe (ABP) specifically binds active tryptase. Recombinant cynomolgus monkey active tryptase was pretreated with a tryptase small molecule inhibitor (10 μM BMS-262084) or 10 μg/mL dissociating anti-tryptase antibody (31A.v11) for 30 min prior to labeling with 10 μM biotinylated-ABP (Bio-PK-DPP (activity-based probe 4) for 20 min. Samples were run on a 4%–12% Bis-Tris denaturing gel and visualized by western blotting with SA-HRP (top panel) to show levels of active tryptase. Blot was stripped and re-blotted (bottom panel) with an antibody that recognizes total tryptase (AA1).