Development of an electrochemiluminescence immunoassay for quantitative determination of semaglutide in human serum.

Semaglutide, a long-acting glucagon-like peptide-1 receptor agonist (GLP-1RA), is widely used in the treatment of patients with type 2 diabetes mellitus and obesity. A sensitive and reliable quantification method is essential for bioanalysis of semaglutide in clinical studies. To develop a sensitive and specific immunoassay, we have prepared a pair of highly specific anti-semaglutide monoclonal antibodies and subsequently employed them as critical reagents in the development of an electrochemiluminescence immunoassay (ECLIA) for the quantification of semaglutide levels in human serum. The assay validation data presented here demonstrate that the performance of the ECLIA method meets prespecified criteria for all validation parameters within the quantitative range of 0.5 ng/mL to 200.0 ng/mL. The intra- and inter-assay precision (%CV) are both ≤14.4%, and accuracy (%RE) ranges from -12.5% to 15.3%. No interference is found with recombinant human GLP-1 as the endogenous counterpart, confirming the assay specificity. Additional assay performance parameters, including selectivity, dilutional linearity, hook effect, and sample stability, all meet the acceptance criteria preset in accordance with international guidelines for bioanalytical method validation. The method is suitable for bioanalysis of semaglutide in human serum.