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Of note is that tirzepatide was previously reported to cause biased signaling at GLP-1R in favor of cAMP response over -arrestin recruitment in the presence of 0.1% casein 39 , which was confirmed in the present study showing that non-acylated tirzepatide elicited better cAMP responses (EC 50 = 0.90 pM for 0.1% casein and 4.66 pM for 0.1% BSA) than that of tirzepatide (EC 50 = 3.05 pM for 0.1% casein and 103.10 pM for 0.1% BSA) (Supplementary Table 11) highlighting the impact of acylation on their pharmacological properties