Anti-Human Hepsin Recombinant Antibody scFv Fragment (Mab2.7.35) (CAT#: TAB-030CT-S(P))

Figure 1 shows a further sensogram of an epitope mapping experiment.

Here anti-hepsin antibody 2.7.35, 2.3.35 and 2.14.5 were immobilized on a CM5 chip and 50 nM hepsin was added. Then antibody 2.7.35 was injected with a concentration of 50 nM.

Figure 2 Comparison of inhibition of hepsin antibodies.

Inhibition of hepsin activity by antibody mouse Mab 2.7.35 (= mH35) (squares), chimeric Mab 2.7.35 (= chH35) (circles) and Mab55 (= hH35) (triangles). After preincubation of antibodies with hepsin for 30 minutes, hydrolysis reaction was started with addition of Ac-KQLR-AMC peptide. Fluorescence increase was measured after 40 minutes incubation. The IC50 values were calculated after fitting the % inhibition data to a four-parameter equation.

Figure 3 Binding of Mab55 (= hH35) to cell surface hepsin.

HEK293 cell lines that stably overexpress full length hepsin with a C-terminal GFP fusion tag were analyzed by flow cytometry. By incubation with increasing amounts of Mab55, specific and saturable surface staining could be detected.

Figure 4 Activity of hepsin antibodies against other serine proteases.

Hepsin (1 nM), Matriptase (2 nM), HAT (2 nM), Enteropeptidase (0.7 nM), Trypsin (1.8 nM) were incubated with antibody (500 nM) for 30 minutes and reaction was started with 5 μΜ Ac-KQLR-AMC peptide.

Figure 5 Activity of antibody Mab55 analyzed in a FRET activity assay against human hepsin (blue), cynomolgus (brown) and mouse (magenta).

Cross reactivity was found against cynomolgus, but not against mouse hepsin.

Figure 6 Progress curve of hepsin inhibition by Mab55 (= hH35).

Hepsin (1 nM) was added to a mixture of Ac-KQLR-AMC peptide (10 μΜ) and 0, 111.1, 333.3 and 100 nΜ of Mab55. The increase of fluorescence was monitored at 10-second intervals over 200 seconds. The amidolytic activity of hepsin in absence of antibody shows a straight line (R² = 0.9946).