Rapid Screening of a Cell-based Assay for GLP-1 Receptor Using a Natural Product Library

Tag-lite GLP-1 binding assay validated on Biotek's Synergy NEO HTS multi-mode Microplate reader.

Larson B, Pierre N, Graham S, Tardieu JL, Degorce F, Banks P


BioTek Instruments, Winooski , VT, USA. Isbio US, Inc., Bedford, MA, USA.

Miptec 2012, Basel, Switzerland / SLAS 2013, Orlando USA

Glucagon-like peptide-1 receptor (GLP-1R) is a G-protein coupled receptor that is present in insulin-secreting beta cells. Intact GLP-1(1-37) is produced by posttranslationalprocessing of proglucagon precursor and converted into active form of GLP-1 [(7-36) amide and (7-37)] with N-terminal truncation. Then, active forms of GLP-1 are deactivated by the further fragmentation with peptidases1,2. Its defining action is augmentation of glucose-induced insulin secretion following intake of carbohydrates and lipids. It also inhibits glucagon secretion and food intake. For this reason, GLP-1R is an interesting target for type-2 diabetes intervention. Here we will demonstrate the detection of GLP-1R binding through the incorporation of a homogeneous cell-based binding assay using Tag-lite® technology. The assay was automated using a non-contact liquid dispenser commonly used in highthroughput settings. Detection of the two fluorescent emissions was accomplishedsimultaneously using the matched PMTs of an HTS multi-mode microplate reader. Optimization experiments were performed to validate the proper conditions to use for automated assay processing. A small primary screen of natural products was then performed under high throughput screening conditions, followed by the creation of dose-response curves using known GLP-1R binders.

GPCR research from A to Z , HTRF® Technology, HTRF microplate readers