Improving sensitivity and drug tolerance of assays for neutralizing anti-drug antibodies to semaglutide and native GLP-1

The purpose of this work was to optimize the sensitivity and the drug tolerance of two cell-based assays for the detection of neutralizing antibodies (nAbs) to semaglutide (a GLP-1 analogue) and to endogenous GLP-1. The two assays were developed and validated in three distinct iterations. Enhancements in sensitivity and drug tolerance were achieved through platform optimization, sample pre-treatment and the alteration of control antibodies. The sensitivity and drug tolerance improved gradually with the different versions of the assays. For detection of nAbs to semaglutide, sensitivity was improved from 3,400 ng to 98 ng/ml antibody, and drug tolerance was improved from 2.5 nM semaglutide when detecting 3,400 ng/ml antibodies to 4.8–5.6 nM semaglutide when detecting 1,000 ng/ml antibody. For the endogenous GLP-1 assay, sensitivity was improved from 6,900 ng/ml to 46 ng/ml antibody, and drug tolerance improved from 1.0 nM semaglutide when detecting antibody concentration of 8,800 ng/ml to 2.5 nM semaglutide when detecting 1,000 ng/ml antibody. Key factors for enhancing the sensitivity and drug tolerance of the assays included the concentration of the drug standard for receptor activation, pre-treatment of the samples, and better understanding of the binding properties of the control antibody. This study aimed to improve two cell-based tests that measure the presence of neutralizing antibodies against semaglutide, a medication similar to glucagon-like peptide-1 (GLP-1), and against the body’s own GLP-1. The researchers created and refined these tests in three different stages. They improved how sensitive the tests were and how much of the drug the tests could handle by optimizing the testing process, preparing samples in specific ways, and changing the control antibodies used. The results showed significant improvements. Overall, the study found that making changes to drug amounts, properly preparing samples, and understanding how control antibodies bind to the drugs were key for these improvements.

本研究旨在优化两种基于细胞的检测方法（cell-based assays）的灵敏度与耐药物性，用于检测针对司美格鲁肽（semaglutide，一种GLP-1类似物（GLP-1 analogue））以及内源性GLP-1的中和抗体（neutralizing antibodies, nAbs）。两种检测方法历经三轮独立迭代完成开发与验证。通过平台优化、样本前处理（sample pre-treatment）以及对照抗体（control antibody）的调整，实现了灵敏度与耐药物性的提升。随着检测方法版本迭代，其灵敏度与耐药物性逐步改善。针对司美格鲁肽中和抗体的检测，灵敏度从3400 ng/ml抗体提升至98 ng/ml；耐药物性则从检测3400 ng/ml抗体时可耐受2.5 nM司美格鲁肽，优化至检测1000 ng/ml抗体时可耐受4.8~5.6 nM司美格鲁肽。针对内源性GLP-1的检测，灵敏度从6900 ng/ml抗体提升至46 ng/ml；耐药物性则从检测8800 ng/ml抗体时可耐受1.0 nM司美格鲁肽，优化至检测1000 ng/ml抗体时可耐受2.5 nM司美格鲁肽。提升该类检测方法灵敏度与耐药物性的关键因素包括受体激活所用药物标准品的浓度、样本前处理流程，以及对对照抗体结合特性的更深入认知。本研究旨在优化两种基于细胞的检测手段，以检测针对司美格鲁肽——一种与胰高血糖素样肽-1（glucagon-like peptide-1, GLP-1）结构相似的药物——以及人体自身GLP-1的中和抗体。研究人员通过三个不同阶段完成了此类检测方法的构建与优化：通过优化检测流程、采用标准化样本制备方式以及调整所用对照抗体，提升了检测的灵敏度与药物耐受能力。实验结果显示检测性能获得了显著改善。综上，本研究证实，调整药物标准品用量、规范样本前处理流程以及深入解析对照抗体的结合特性，是实现上述性能提升的关键因素。