Fig 1: Pharmacological characterization of GLP1R, GIPR and GCGR binders.a, Antagonism of miniproteins dGI1_024 and mGI1_008 at the GLP1R were tested using the reporter cell line BHK21/GLP1R/Cre-luc. Cells were treated with varying concentrations of miniproteins prior to treatment with 15 pM of semaglutide, a GLP1R agonist. Both binders antagonized GLP1R signaling with IC50 values of 61 ± 20 nM for dGI1_024 and 39 ± 18 nM for mGI1_008. Exendin 9–39 was used as a positive control and had an IC50 13.9 ± 1.07 nM. Data are shown as mean ± SD (n=2). Binding affinity of miniprotein binders targeting b GIPR and c GCGR was measured by surface plasmon resonance (SPR). SPR sensorgram of binders with the highest affinity is shown. Computational design models are highlighted in blue and the target receptor in yellow.