Recombinant Human Anti-Nicotine Antibody (Nic12)

(A) Inhibition of nicotine entry into brain. Mice were injected i.p. with 0.5 mg of the indicated human IgG2. One day later, mice were injected i.v. with 750 ng of tritium-labeled
nicotine. Mice were killed after 5 min and nicotine concentrations in brains were measured. Individual mice are shown; bars indicate average values. IgG, control human IgG. *, P < 0.001 vs. IgG; **, P < 0.003 vs. IgG. (B) Dose-response of inhibition of nicotine entry into brain by IgG2-Nic12. Mice were injected i.p. with 0.1, 0.3, 1, or 3mg of mAb-Nic12. One day later, mice were injected i.v. with 750 ng of tritium-labeled nicotine. Mice were killed after 5 min and nicotine concentrations in brains were measured. Antibody concentrations in sera were determined and plotted against the percentage nicotine-reduction in the brain. (C) Time course of inhibition of nicotine entry into brain by IgG2-Nic12. Mice were injected with 0.5mgantibody, challenged with750ngof tritium-labeled nicotine and nicotine in the brain was measured at the indicated times. Averages of 4 to 5 mice are given with standard deviations. IgG, control human IgG. *, P < 0.001 vs. IgG. (D) Specificity of mAb-Nic12. The binding of tritium-labeled-nicotine (56 nM) to mAb-Nic12 (50 nM) was measured by equilibrium dialysis in the presence of increasing concentrations of unlabeled (-)-nicotine, (-)-cotinine, or acetylcholine. Averages of two independent experiments are given with standard deviations.

Beerli, R. R., Bauer, M., Buser, R. B., Gwerder, M., Muntwiler, S., Maurer, P., ... & Bachmann, M. F. (2008). Isolation of human monoclonal antibodies by mammalian cell display. Proceedings of the National Academy of Sciences, 105(38), 14336-14341.