Method for detecting binding force of monoclonal antibody to RANKL and application thereof

• Main Authors: YIN XIAOJIE, DAI HU, TIAN ZHENZHEN, HAN ARONG, DING MANSHENG, SHEN HONG
• Format: Patent
• Language: Chinese; English
• Published: 25.04.2023
• Subjects: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIRCHEMICAL OR PHYSICAL PROPERTIES; MEASURING; PHYSICS; TESTING

Aiming at the technical problems that an indirect ELISA method for detecting the antigen binding activity of an antibody in the preparation process of an anti-RANKL monoclonal antibody is complex in operation, multiple in cascade amplification steps and time-consuming and labor-consuming in plate washing, after a to-be-detected antibody is added for plate washing, a universal secondary antibody and Multi-Species SP ads-HRP are synchronously added, the cascade amplification step and the signal molecule introduction step are combined, and the detection sensitivity is high. The tedious operation caused by layer-by-layer cascade signal amplification is reduced, and the accuracy and repeatability of a detection result are ensured. 本发明针对抗RANKL单克隆抗体制备过程中用于检测抗体对抗原结合活性的间接ELISA法操作复杂、级联放大步骤多、洗板费时费力的技术问题，在加入待测抗体洗板后，同步加入通用二抗和Multi-Species SP ads-HRP，将级联放大步骤和信号分子引入步骤合并，不仅减少了逐层级联放大信号导致的繁琐操作、而且确保了检测结果的准确度和重复性。