Anti-Human CD14 Recombinant Antibody (rl8D11) (CAT#: TAB-1585CL)

Figure 1 The Ab concentration in cell culture supernatant (mg/ml; s) and the Ab production rate (pg/cell/day; d) during a fed-batch expression period of 12 d were determined with ELISA.

Data are given as mean and SEM for rMil2 (A; n = 12), r18D11 (B; n = 5) and raNIP (C; n = 2).

Lau, C., Gunnarsen, K. S., Høydahl, L. S., Andersen, J. T., Berntzen, G., Pharo, A.,... & Nielsen, E. W. (2013). Chimeric anti-CD14 IGG2/4 hybrid antibodies for therapeutic intervention in pig and human models of inflammation. The Journal of Immunology, 191(9), 4769-4777.

Figure 2 Functional characterization of anti-human CD14 Ab r18D11.

(A) Binding of increasing concentrations of r18D11 (△), raNIP (□), 18D11 F(ab)′2 (▴) or a control F(ab)′2 (▪) to monocytes was determined by the ability of the Abs to displace 10 μg/ml of the original clone 18D11 mIgG1 from its CD14 binding site in human whole blood. Mean fluorescence intensity derived from a PE-conjugated anti-mouse IgG Ab was measured using flow cytometry. Fluorescence intensity in presence of original clone 18D11 alone was set to 100%. (B-D) Release of the proinflammatory cytokines IL-1β (B), TNF (C), and IL-6 (D) from human whole blood was induced with 100 ng/ml ultrapure LPS from E. coli O111:B4 in the presence of increasing concentrations of r18D11 (△) or the original clone 18D11 (▴). raNIP (□) and mIgG1 isotype control (▪) (10 μg/ml) served as negative controls. (E) Monocyte oxidative burst was measured with flow cytometry after adding the different Ab preparations to human whole blood. Data are given as mean and SEM (n = 3 independent experiments). *p < 0.05 as compared with the negative PBS control.

Lau, C., Gunnarsen, K. S., Høydahl, L. S., Andersen, J. T., Berntzen, G., Pharo, A.,... & Nielsen, E. W. (2013). Chimeric anti-CD14 IGG2/4 hybrid antibodies for therapeutic intervention in pig and human models of inflammation. The Journal of Immunology, 191(9), 4769-4777.

Figure 3 In vitro binding of recombinant IgG2/4 hybrid Abs to complement and Fc-receptors.

Increasing concentrations of rMil2 (s), r18D11 (O) or raNIP (N) were incubated with (A) immobilized human C1q, (B) the human Fcg receptors FcgRI, (C) FcgRIIa (allotype His131), (D) FcgRIIb, (E) FcgRIIIa (allotype Val158), (F) FcgRIIIb, and (G, H) human (hFcRn) or (I, J) porcine FcRn (pFcRn) at acidic (pH 6.0) and neutral pH (pH 7.4). Human IgG1 ()) (A-H) or porcine IgG (pIgG; ♦) (I, J) served as positive controls. Binding was measured as absorbance at 450 nm. Data are given as mean and SD (n = 3 independent experiments).

Lau, C., Gunnarsen, K. S., Høydahl, L. S., Andersen, J. T., Berntzen, G., Pharo, A.,... & Nielsen, E. W. (2013). Chimeric anti-CD14 IGG2/4 hybrid antibodies for therapeutic intervention in pig and human models of inflammation. The Journal of Immunology, 191(9), 4769-4777.