Division of Infection Medicine, Department of Clinical Sciences Lund, Faculty of Medicine, Lund University, Lund, Sweden.
Methods Mol Biol. 2023;2674:251-259. doi: 10.1007/978-1-0716-3243-7_17.
Antibody binding to bacterial surfaces plays a crucial role in immunity, and a key characteristic of this protein-protein interaction is the binding affinity. Determining the affinity of an antibody binding to its antigen is the first step in predicting the function in a physiological environment where other competing protein interactions may be present. Antibody-antigen affinity is often evaluated with isolated proteins. It is informative to also be able to assess antibody binding to a bacterial surface where many antigens might be present, including multiple copies of the specific antigen the antibody recognizes, and in a context where the antigen might be in a more natural conformation. In this chapter, we present a flow cytometry-based assay to measure and calculate the cell surface binding affinity or avidity of any mono- or polyclonal antibody solution.
抗体与细菌表面的结合在免疫中起着至关重要的作用，这种蛋白质-蛋白质相互作用的一个关键特征是结合亲和力。确定抗体与其抗原的结合亲和力是预测在存在其他竞争蛋白相互作用的生理环境中功能的第一步。抗体-抗原亲和力通常使用分离的蛋白质进行评估。能够评估抗体与细菌表面的结合也很有意义，因为在这种情况下，可能存在许多抗原，包括抗体识别的特定抗原的多个拷贝，并且抗原可能处于更自然的构象。在本章中，我们介绍了一种基于流式细胞术的测定法，用于测量和计算任何单克隆或多克隆抗体溶液的细胞表面结合亲和力或亲合力。
