摘要
采用生物膜层干涉技术建立了一种准确评估单克隆抗体与Fc受体结合力的方法。通过优化关键参数,包括配体固化浓度、高度、结合时间和分析物浓度,确定了最佳的实验条件:配体固化浓度为10μg/mL,固化高度控制低于0.5 nm,单克隆抗体与Fc受体结合时间为90 s,检测分析物浓度低于100 nmol/L。实验结果显示,该方法能形成良好的结合解离曲线,不同浓度范围内呈现明显的跨度,并且具有良好的实验重复性。这种无需标记的单克隆抗体与Fc受体结合力检测方法,对于研究蛋白相互作用以及复杂生物样品的检测具有重要意义。
A precise method for evaluating the binding affinity between monoclonal antibodies and Fc receptors has been established using biolayer interferometry(BLI)technique.By optimizing key parameters,including ligand immobilization concentration,height,binding time,and analyte concentration,the optimal experimental conditions were determined:a ligand immobilization concentration of 10μg/mL,immobilization height controlled below 0.5 nm,a monoclonal antibody-Fc receptor binding time of 90 s,and analyte concentration detection below 100 nmol/L.Well-defined binding and dissociation curves were formed by this method,with significant spans observed across different concentrations,demonstrating excellent experimental reproducibility.This label-free method for assessing the binding affinity between monoclonal antibodies and Fc receptors holds great importance in studying proteinprotein interactions and analyzing complex biological samples.
作者
陈玉茹
罗明
陈涛
张甲
颜娟
李兰英
Chen Yuru;Luo Ming;Chen Tao;Zhang Jia;Yan Juan;Li Lanying(College of Food Science and Technology,Shanghai Ocean University;Key Laboratory of Bioanalysis and Metrology for State Market Regulation,Shanghai Institute of Measurement and Testing Technology;Sartorius(Shanghai)Trading Co.,Ltd.)
出处
《上海计量测试》
2024年第3期2-6,10,共6页
Shanghai Measurement and Testing
基金
国家质量基础设施体系专项(2021YFF0600705)
上海市市场监督管理局科技项目(2022-13)。
关键词
单克隆抗体
生物膜层干涉法
结合力
传感器
monoclonal antibody
biolayer interferometry
binding affinity
biosensor
