NeuroMab™ Anti-Transferrin Receptor Antibody(NRP-0422-P1313)

Figure 1 Inhibition of protease-mediated PCSK9 cleavage by mutagenesis of "loop 218" residues and antibody 3D5.

PCSK9 (1.9 μΜ) was preincubated with antibodies 3D5 or 7G7 for 20 min before treatment with 40 nM hepsin or with 80 nM furin for 6 h. Results indicate that antibody 3D5 completely inhibited PCSK9 cleavage by either protease, whereas antibody 7G7 did not.

Figure 2 Inhibition of PCSK9 function by Ab-3D5.

Inhibition of PCSK9 binding to LDLR by biolayer interferometry. LDLR ectodomain was immobilized on the sensor of an Octet Red384 system (ForteBio; Menlo Park, CA) and binding to PCSK9 alone (Ctrl) or preincubated with Ab-3D5 or the non-blocking Ab-7G7 was measured. Results are the average of n=3 ± SD.

Figure 3 Inhibition of PCSK9 function by Ab-3D5.

Ab-3D5 prevents LDLR degradation in HepG2 cells. HepG2 cells were treated for 4 h with buffer alone (Ctrl), with PCSK9 alone (-Ab) or with PCSK9 preincubated with increasing concentrations of the non-blocking Ab-7G7 or with Ab-3D5. Surface LDLR levels were quantified by FACS analysis and expressed as percent of control levels.

Figure 4 shows the effect of antibody 3D5 on the activity of intact and hepsin-cleaved PCSK9 in the HepG2 assay.

Figure 5 shows that antibody 3D5 neutralizes PCSK9 activity in a mouse model. Mice received 20 mg/kg antibody two hours before injection of 30 μg PCSK9 1 hour.