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Subsequently, the binding affinity of native semaglutide and GLP-1R ECD was calculated by Prodigy [45, 46] to be 3.4 10 6 M at 37 C, with the template being PDB entry: 4ZGM [29], where a total of 42 residue pairs were found to be located at the binding interface of semaglutide and GLP-1R ECD [29, 52], as listed in supplementary file 4zgm.ic