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Following cleavage with Precision Protease (Beyotime) to remove the GST tag, the eluted proteins underwent further purification by size-exclusion chromatography on a Superdex 200 10/300 GL column (GE Healthcare) using an elution buffer of 50 mM HEPES (pH 7.4) and 150 mM NaCl

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The comparator arm was placebo, a GLP-1 RA (subcutaneous administration of semaglutide 1 mg once-weekly) and basal insulin in three [18, 19, 22], one [24, 25] and two trials [20, 21], respectively, while one trial included both a placebo arm and a GLP-1 RA arm (dulaglutide 1.5 mg once-weekly) [23]