Department of Bioengineering, School of Engineering, The University of Tokyo, Tokyo, Japan.
AIDS Research Center, National Institute of Infectious Diseases, Tokyo, Japan.
Methods Mol Biol. 2023;2552:409-433. doi: 10.1007/978-1-0716-2609-2_23.
In the computational design of antibodies, the interaction analysis between target antigen and antibody is an essential process to obtain feedback for validation and optimization of the design. Kinetic and thermodynamic parameters as well as binding affinity (K) allow for a more detailed evaluation and understanding of the molecular recognition. In this chapter, we summarize the conventional experimental methods which can calculate K value (ELISA, FP), analyze a binding activity to actual cells (FCM), and evaluate the kinetic and thermodynamic parameters (ITC, SPR, BLI), including high-throughput analysis and a recently developed experimental technique.
在抗体的计算设计中，目标抗原与抗体之间的相互作用分析是获得设计验证和优化反馈的必要过程。动力学和热力学参数以及结合亲和力 (K) 可以更详细地评估和理解分子识别。在本章中，我们总结了可以计算 K 值的常规实验方法（ELISA、FP）、分析与实际细胞的结合活性（FCM）以及评估动力学和热力学参数（ITC、SPR、BLI），包括高通量分析和最近开发的实验技术。
