The Establishment of GLP-1 Analogue Detection Cell Model and its Functional Analysis

Abstract: In order to explore the factors influencing the stability and repeatability of the activity of GLP-1 analogue assay by cell models， a eukaryotic expression vector pEGFP-N1/hGLP-1R-EGFP was constructed and transfected into Chinese hamster ovary (CHO) cells. After limited selection by G418， the combination selection of flow cytometry and limiting dilution was performed and a stable and highly expressed monoclonal cell line was obtained. Inverted fluorescence analysis， flow cytometry analysis and RT-PCR results showed that the gene was transcribed and translated， and the expressed receptor protein was located on the cell membrane side. Liraglutide and Exendin-4 were used as a model drug for the activity assay， respectively. The results showed that the cell model has high drug sensitivity， relatively stable activity assay with different model cell generations and initially explored the receptor expression of model cells， fetal bovine serum and its single component BSA has a significant effect on the drug EC50 measurements. In summary， the established cell model can be well applied to the activity test of GLP-1 analogue， providing a simple and reliable model for the screening and activity evaluation of GLP-1 analogues with good activity stability and repeatability.

CSTR: 32200.14.cjcb.2018.06.0016