Human Anti-APS Recombinant Antibody (clone CL1) (CAT#: HPAB-1960-FY)

Figure 1 Four monoclonal aCL bind to human β2GP1 alone.

Antibodies were used at 1 mg/ml (filled and open bars) or 10 mg/ml (slashed bars; denoted Ab 10X), except for CL15 and CL24 (which were used at 2 mg/ml and 20 mg/ml, respectively).

Zhu, M. , Olee, T. , Le, D. T. , Roubey, R. A. S. , Hahn, B. H. , & Woods, J. V. L. , et al. (1999). Characterization of igg monoclonal anti-cardiolipin/anti-beta2gp1 antibodies from two patients with antiphospholipid syndrome reveals three species of antibodies. British Journal of Haematology, 105(1), 102-109.

Figure 2 Characterization of the serum cofactors for monoclonal aCL.

All antibodies were used at 1 mg/ml, except for CL15 and CL24 which were used at 2 mg/ml.

Zhu, M. , Olee, T. , Le, D. T. , Roubey, R. A. S. , Hahn, B. H. , & Woods, J. V. L. , et al. (1999). Characterization of igg monoclonal anti-cardiolipin/anti-beta2gp1 antibodies from two patients with antiphospholipid syndrome reveals three species of antibodies. British Journal of Haematology, 105(1), 102-109.

Figure 3 Effect of anti-β2-glycoprotein I (anti-β2GPI) IgG on the release of neutrophil extracellular traps (NETs) by neutrophils.

NET release from neutrophils stimulated with the indicated antibodies. Control neutrophils were treated with purified monoclonal antiphospholipid antibodies (aPL; 10 mg/ml) for 3 hours in the presence or absence of 10% autologous serum. IS4, aPL, CL1, and CL24 are known to bind b2GPI, while IS1 and IS2 do not.

Yalavarthi, S., Gould, T. J., Rao, A. N., Mazza, L. F., Morris, A. E., Núñez‐Álvarez, C., ... & Knight, J. S. (2015). Release of neutrophil extracellular traps by neutrophils stimulated with antiphospholipid antibodies: a newly identified mechanism of thrombosis in the antiphospholipid syndrome. Arthritis & rheumatology, 67(11), 2990-3003.

Figure 4 Stimulation of thrombin generation by purified antiphospholipid antibodies (aPL), as well as plasma from patients with antiphospholipid syndrome (APS), in a neutrophil- and DNA-dependent manner.

Representative thrombin (IIa) generation plot demonstrating enhanced generation when neutrophils are exposed to platelet-poor plasma (from a healthy control) supplemented with aPL CL1 (10 mg/ml).

Yalavarthi, S., Gould, T. J., Rao, A. N., Mazza, L. F., Morris, A. E., Núñez‐Álvarez, C., ... & Knight, J. S. (2015). Release of neutrophil extracellular traps by neutrophils stimulated with antiphospholipid antibodies: a newly identified mechanism of thrombosis in the antiphospholipid syndrome. Arthritis & rheumatology, 67(11), 2990-3003.

Figure 5 Blocking of antiphospholipid antibody (aPL)-mediated neutrophil extracellular trap (NET) release by inhibition of Toll-like receptor 4 (TLR-4) but not TLR-2.

Yalavarthi, S., Gould, T. J., Rao, A. N., Mazza, L. F., Morris, A. E., Núñez‐Álvarez, C., ... & Knight, J. S. (2015). Release of neutrophil extracellular traps by neutrophils stimulated with antiphospholipid antibodies: a newly identified mechanism of thrombosis in the antiphospholipid syndrome. Arthritis & rheumatology, 67(11), 2990-3003.

Figure 1 Human Anti-APS Recombinant Antibody (clone CL1) (HPAB-1960-FY) in SDS-PAGE

SDS-PAGE analysis of HPAB-1960-FY in reduced (Lane 1) and non-reduced (Lane 2) conditions. Gel stained for 30 minutes with Coomassie Blue. As a result, different β-mercaptoethanol-reduced proteins (Heavy chain and Light chain) migrate as about 50 kDa and 25 kDa, respectively.

The purity of HPAB-1960-FY was greater than 95% as determined by SDS-PAGE.

Figure 2 Human Anti-APS Recombinant Antibody (clone CL1) (HPAB-1960-FY) in HPLC

The purity of HPAB-1960-FY was greater than 95% as determined by SEC-HPLC.