NeuroMab™ Anti-PADI2 BBB Shuttle Antibody, Clone #33

Figure 1. The culture supernatant of mAb #1-35 was tested on human recombinant (hr) PAD2-coated plates.

hrPAD2 was diluted in 2-fold steps from 500 ng/mL. Shown is the absorbance at the coating concentration of 32 ng/mL. HRP-conjugated rabbit anti-mouse, diluted 1:1000, was added for 1 hour at RT, and the plates were developed with OPD substrate. The levels are given as OD490-650 nm-units.

Figure 2. mAbs reacted with hrPAD2 in western blotting.

Figure 3. Culture supernatant from mAb #1-35 tested on hrPAD2/hrPAD4 coated plates - 50 ng/mL.

HRP Rabbit anti-mouse (p0260) was added 1:1000 for 1 hour at RT and plates were developed with OPD substrate. The levels are given as OD490-650 nm-units.

Figure 4. Anti-PAD2 epitope mapping. Different splice variants of PAD2 were assessed by western blotting.

Shown is the reactivity of three selected mAbs (#2, #6 and #34) with WT (full length wild type human PAD2), C254 (amino acids 1-254 of human PAD2), 1385-463 (whole length human PAD2 without the catalytic site), N165 (from amino acid 165 to the C-terminus), N343 (from amino acid 343 to the C-terminus). The mAbs were found to bind in the N-terminal region among amino acids 1-165.

Figure 5. Inhibitory ability of anti-PAD2 monoclonal antibody. The ability of selected anti-PAD2 mAbs to inhibit fibrinogen citrullination was tested using human recombinant PAD2 (hrPAD2) as a catalyst.

Test of the inhibitory capacity of mAbs #2, #3 and #33. A mAb against human complement component 4 C4 (anti-C4) was used as negative control.

Figure 6. Inhibitory ability of anti-PAD2 mAb. The ability of selected anti-PAD2 mAbs to inhibit fibrinogen citrullination was tested using human recombinant PAD2 (hrPAD3) as a catalyst.

Test of the inhibitory capacity of mAbs #6, #8 and #10. Anti-C4 was used as control.

Figure 7. Inhibitory ability of anti-PAD2 mAb. The ability of selected anti-PAD2 mAbs to inhibit fibrinogen citrullination was tested using human recombinant PAD2 (hrPAD4) as a catalyst.

Test of the inhibitory capacity of culture supernatants (cs) from mAb #9, #12, #31 and #34 was tested. mAbs against chicken complement component 3 (chC3) and SCUBE1 (signal peptide, CUB domain, epidermal growth factor-like protein 1) was used as controls.