Fig. 1: Development of mGlyR-selective nanobodies.

a Schematic of the nanobody development pipeline. A phage library was constructed from leukocytes of a llama immunized with mGlyR, followed by 3 rounds of panning on mGlyR to enrich for specific binders, which were isolated and tested. b Schematic of the detection strategy in flow cytometry experiments. c Analysis of nanobody binding by flow cytometry of HEK cells transiently expressing mGlyR incubated with or without Nb20 and anti-myc-APC conjugated antibody. Percentages of cells in each quadrant are indicated. d Dose-response profiles of representative Nb20 binding experiment to cells expressing mGlyR in flow cytometry experiments. Concentrations of Nb20 are shown. e Quantification of data in panel D. Error bars are SEM values (n = 3 independent experiments). f Schematic of the surface plasmon resonance (SPR) assays that detect Nb20 binding to the chip containing immobilized recombinant ectodomain of mGlyR (Ecto-mGlyR). g SPR sensorgram of binding and dissociation of Nb20.