Human Anti-GIPR Recombinant Antibody (clone Gipg013) (CAT#: PABL-103)

Figure 1 Antagonism of GIP induced cAMP production in GIPr over-expressing cell lines.

Gipg013 and Gipg133 were characterised for GIPr antagonism in cell based cAMP HTRF® assay and data plotted using nonlinear regression. Panel A and B show antagonistic profiles from HEK293s cells overexpressing human, mouse, rat, and, dog GIPr using Gipg013 and Gipg133 IgGs, respectively,. Panel C and D show antagonistic and agonistic profiles in the human GIPr assay for GIP7-30 and Pro3GIP and GIP. Values have been normalised to the maximum activity of GIPr, which is defined by total cellular cAMP produced in the agonism assay on in the absence of peptide/IgG in the antagonism assay. ●, human; ■, mouse; ▼, rat; ▲, dog; ♦, isotype control IgG1; *, GIP; +, GIP7-30; ×, Pro3GIP.

Ravn, P., Madhurantakam, C., Kunze, S., Matthews, E., Priest, C., O'brien, S.,... & Benthem, L. (2013). Structural and pharmacological characterisation of novel potent and selective monoclonal antibody antagonists of glucose-dependent insulinotropic polypeptide receptor. Journal of Biological Chemistry, jbc-M112.

Figure 2 Antibody cell binding and inhibition of ligand binding.

A, Binding of Gipg013 to GIPr and related receptors on overexpressing cells. Direct binding of 0.25 µg/ml Gipg013 IgG to h, human; d, dog; m, mouse; or r, rat orthologs of the glucagon (GCGr), GLP-1 (GLP-1r) or GIP (GIPr) receptors. Control for non-specific binding on GIPr orthologs, NIP228_TM, at 0.25 µg/ml. Data shown is representative of two separate experiments for the human receptors and a single experiment for the rodent and canine receptors. B, receptor ligand competition assay showing IC50 determination of Gipg013 IgG binding to GIPr overexpressing cells. □, Gipg013; ▲, isotype control IgG1.

Ravn, P., Madhurantakam, C., Kunze, S., Matthews, E., Priest, C., O'brien, S.,... & Benthem, L. (2013). Structural and pharmacological characterisation of novel potent and selective monoclonal antibody antagonists of glucose-dependent insulinotropic polypeptide receptor. Journal of Biological Chemistry, jbc-M112.

Figure 3 Analysis of Gipg013 antagonism of GIPr.

A, GIP dose response curves in the presence of Gipr013. Non linear regression plot of dose effect curve for GIP was determined in the presence of various concentrations of Gipg013 IgG. ∇, 3750 nM; ∆, 1250 nM; □, 417 nM; ○, 139 nM; ♦, 46 nM; ▼15 nM; ▲, 5 nM; ■, 1.7 nM; ●, 0 Gipg013. B, Schild plot analysis of dose response curves. The Schild plot intersects the abscissa at pA2 (=KD).

Ravn, P., Madhurantakam, C., Kunze, S., Matthews, E., Priest, C., O'brien, S.,... & Benthem, L. (2013). Structural and pharmacological characterisation of novel potent and selective monoclonal antibody antagonists of glucose-dependent insulinotropic polypeptide receptor. Journal of Biological Chemistry, jbc-M112.

Figure 4 Inhibition of GIP induced insulin secretion.

Panels A and B show inhibition of GIP stimulation in a glucose stimulated insulin secretion assay with dispersed rat islets. A, The the effect of GIP on top of the glucose stimulated insulin secretion (open bars) and the inhibition of this effect with 600 nM Gipg013 (filled bars). B, The GIP enhanced insulin secretion was inhibited with Gipg013 at concentrations of both 300 nM and 600 nM. Panel C shows Inhibition of GIP induced insulin secretion in vivo. Insulin response measured as the relative insulin AUC0-30 min between period one and two for GIP induced insulin secretion in anesthetized rats, AUC1 before and AUC2 30 minutes after Gipg013 infusion. Insulin response = AUC2/AUC1*100. n=6 for vehicle and n=3 for Gipg013. *p<0.05 between vehicle and Gipg013.

Ravn, P., Madhurantakam, C., Kunze, S., Matthews, E., Priest, C., O'brien, S.,... & Benthem, L. (2013). Structural and pharmacological characterisation of novel potent and selective monoclonal antibody antagonists of glucose-dependent insulinotropic polypeptide receptor. Journal of Biological Chemistry, jbc-M112.