Development and verification of a method for determining the binding affinity of human serum albumin with semaglutide based on surface plasmon resonance technique
10.13200/j.cnki.cjb.004719
- VernacularTitle:基于表面等离子共振技术的人血白蛋白与司美格鲁肽结合亲和力检测方法的建立及验证
- Author:
Fan WU
- Publication Type:Journal Article
- Keywords:
Surface plasmon resonance(SPR) technique;
Human serum albumin(HSA);
Glucagon-like peptide-1(GLP-1);
Semaglutide
- From:
Chinese Journal of Biologicals
2026;39(06):728-735
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish and verify a surface plasmon resonance(SPR) technique for determining the binding affinity of human serum albumin(HSA) with glucagon-like peptide-1(GLP-1) analog semaglutide,in order to provide a reli-able detection method for the initial screening and quality control of GLP-1 analog drug development.Methods HSA was immobilized on the surface of the CM5 chip using the amino coupling method,semaglutide was injected into the chip channel at a constant flow rate,and the coupling amount(7 800,670 RU),buffer composition(1 × HBS-EP~+,1 × HBS-N,1 × PBS,1 ×PBS + 5% DMSO) and temperature(25 and 37 ℃) were optimized.The specificity,precision and robustness of the method were verified.The established method was used to detect the binding affinity of semaglutide produced by two manufacturers to HAS.Results The optimal coupling amount was 670 RU,the buffer was 1 × PBS + 5% DMSO,and the temperature was25 ℃.The established method could specifically identify the binding effect of semaglutide and HSA,and effectively elimi-nate the interference of irrelevant components such as matrix and buffer.The CVs of precision and robustness verification were both less than 10%.The established method was used to detect the mean value of binding affinity constants(KD) of semaglutide produced by two manufacturers and HSA,which were 2.418 × 10~(-6)and 2.864 × 10~(-6)μmol/L,respectively,and the CVs were both less than 10%.Conclusion The established method based on SPR technology for detecting the binding affinity of HSA and semaglutide has good specificity,precision and robustness,and can be used for the research related to the development of GLP-1 analog drugs.
