Xu Yan, Pan Yuli, Li Lingnuo, Zhou Ming
College of Bioengineering, Chongqing University, Chongqing 400044, P. R. China.
Shenzhen Lifotronic Technology Co., Ltd., 1008 Songbai Road, Shenzhen, Guangdong 518055, P. R. China.
ACS Omega. 2020 Dec 11;5(50):32591-32596. doi: 10.1021/acsomega.0c04877. eCollection 2020 Dec 22.
The capability of biotinylated molecules to bind streptavidin may be a more functional measure of the success of target biotinylation than titration of total bound biotins per molecule. It was demonstrated that the binding capability could be assessed by a competitive assay, in which a biotinylated antibody (BA) (or protein, ligand, receptor etc.) of interest competed with a reference antibody (or a protein) dually labeled with biotin and electrochemiluminescence (ECL) moieties for the binding sites of streptavidin coated on the surface of magnetic beads. Inversely related to the ECL signal, the binding capability of a biotinylated antibody can be reproducibly evaluated by multiple sets of easily acquired data series rather than by a single measurement. This method can be employed in an ordinary laboratory with an automated ECL analyzer or other readout instruments for routine characterization of any biotinylated species, such as proteins, ligands, receptors, and polypeptides.
与滴定每个分子结合的生物素总量相比，生物素化分子与链霉亲和素结合的能力可能是衡量目标生物素化成功与否的更具功能性的指标。结果表明，结合能力可以通过竞争性测定来评估，其中感兴趣的生物素化抗体（BA）（或蛋白质、配体、受体等）与双标记有生物素和电化学发光（ECL）部分的参考抗体（或蛋白质）竞争结合包被在磁珠表面的链霉亲和素的结合位点。与ECL信号呈负相关，生物素化抗体的结合能力可以通过多组易于获取的数据系列进行可重复评估，而不是通过单次测量。该方法可在配备自动ECL分析仪或其他读出仪器的普通实验室中用于常规表征任何生物素化物种，如蛋白质、配体、受体和多肽。
