NeuroMab™ Anti-CD14 BBB Shuttle Antibody(NRZP-1022-ZP3701)

Figure 1 shows the expression of recombinant IgG2/4 hybrid antibody.

(AC) Ab concentration (mg/ml; s) and Ab productivity (pg/cell/day; d) in cell culture supernatants were determined by ELISA during 12 days of fed-batch expression. Data are presented as mean and SEM for rMil2 (A; n=12), r18D11 (B; n=5) and raNIP (C; n=2). (D) 500 ng of each purified antibody was subjected to reducing or non-reducing SDS-PAGE and stained with Coomassie blue. (E) Non-reducing SDS-PAGE and immunoblotting of the same sample (10 ng) using an Ab specific for the human IgG2 hinge region.

Figure 2 shows the functional characterization of anti-human D14 Ab r18D11.

(A) Binding of increasing concentrations of r18D11 (O), raNIP (N), 18D11 F(ab)92 (▴) or control F(ab)92 (n) to monocytes was determined by the ability of Abs to displace 10 mg/ ml Original clone 18D11 mIgG1, CD14 binding site from human whole blood. (BD) Using 100 ng/ml ultrapure LPS from E. coli O111:B4 at increasing concentrations of r18D11 (O) or the original clone 18D11 (▴). (E) Flow cytometry measurement of monocyte oxidative burst after addition of different Ab preparations to human whole blood.

Figure 3 shows the in vitro binding of recombinant IgG2/4 hybrid antibodies to complement and Fc receptors.

Increasing concentrations of rMil2 (s), r18D11 (0) or raNIP (N) were mixed with (A) immobilized human C1q, (B) human Fcg receptor FcgRI, (C) FcgRIIa (allotype His131), (D) FcgRIIb, (E) FcgRIIIa (allotype Val158), (F) FcgRIIIb and (G, H) human (hFcRn) or (I, J) porcine FcRn (pFcRn), acidic (pH 6.0) and neutral pH (pH 7.4).