Imamura Hiroshi, Honda Shinya
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan; Department of Applied Chemistry, College of Life Sciences, Ritsumeikan University, Kusatsu, Japan.
Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan.
Methods Enzymol. 2019;622:329-345. doi: 10.1016/bs.mie.2019.02.021. Epub 2019 Mar 12.
Antibody unfolding and aggregation have been gaining increasing concern as the need for pharmaceutical antibodies with high quality has increased. Assays for continuous application of severe stress to antibodies, which are typically employed to evaluate tolerance to aggregation or to understand the mechanism of aggregation, induce both unfolding and aggregation simultaneously. In this chapter, we describe the pH-shift stress on antibodies that uses a low pH solution to unfold the antibody and a neutralizing solution to induce folding and the aggregation. This pH-shift technique with light scattering analysis will be a useful assay to evaluate antibody aggregation at a neutral pH and ambient temperature without generation of unfolded molecules during aggregation.
随着对高质量药用抗体需求的增加，抗体的解折叠和聚集越来越受到关注。用于持续对抗体施加严苛压力的检测方法，通常用于评估抗体对聚集的耐受性或理解聚集机制，会同时诱导抗体解折叠和聚集。在本章中，我们描述了针对抗体的pH值变化应激，该方法使用低pH溶液使抗体解折叠，并使用中和溶液诱导折叠和聚集。这种结合光散射分析的pH值变化技术，将成为一种有用的检测方法，可在中性pH值和环境温度下评估抗体聚集情况，且在聚集过程中不会产生解折叠分子。
