Kim Dae Young, Ding Wen, Tanha Jamshid
Institute for Biological Sciences, National Research Council Canada, Ottawa, ON, Canada.
Methods Mol Biol. 2012;911:355-72. doi: 10.1007/978-1-61779-968-6_21.
Solubility and stability are amongst the factors contributing to the therapeutic efficacy of biologics. Human antibody heavy chain variable domains, VHs, are one class of biologics; improving VH biophysical properties is the focus of significant protein engineering efforts. Here, we describe an efficacy engineering approach which involves the introduction of a disulfide linkage in the VH core and which improves both VH solubility and stability. More specifically, we describe protocols for generation of disulfide engineered human VHs and their characterization in terms of disulfide linkage formation, non-aggregation, and stability. Our solubility/stability engineering approach may be applied to other VHs.
溶解度和稳定性是影响生物制品治疗效果的因素之一。人源抗体重链可变区（VH）是一类生物制品；改善VH的生物物理特性是大量蛋白质工程研究工作的重点。在此，我们描述了一种效能工程方法，该方法涉及在VH核心区域引入二硫键，从而提高VH的溶解度和稳定性。更具体地说，我们描述了生成二硫键工程化人源VH的方案，以及根据二硫键形成、非聚集性和稳定性对其进行表征的方法。我们的溶解度/稳定性工程方法可应用于其他VH。
