Improved Solution-State Properties of Monoclonal Antibodies by Targeted Mutations
Improved Solution-State Properties of Monoclonal Antibodies by Targeted Mutations
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DOI：
10.1021/acs.jpcb.7b09126
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发表时间：
2017-12-07
影响因子：
3.3
通讯作者：
Schaefer, Lars V.
中科院分区：
文献类型：
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作者：
Kuhn, Alexander B.;Kube, Sebastian;Schaefer, Lars V.
基于单克隆抗体（mAb）的治疗剂通常需要高浓度制剂。不幸的是，高度浓缩的抗体溶液通常具有不利于治疗开发的生物物理性质，例如高粘度、溶解度限制、沉淀问题或液液相分离。在这项工作中，我们提出了一个计算合理的设计原则，通过有针对性的点突变提高单克隆抗体解决方案的热力学稳定性。两种在高浓度下表现出高粘度的公开可得的IgG1单克隆抗体被用作模型系统。通过结合分子动力学模拟与三维参考相互作用位点模型理论的计算有效的方法的指导下，在可变Fv区域中引入带电残基的点突变，以这种方式优化水合自由能。然后通过瞬时表达产生两个选择的点突变体并进行实验表征。与相应的野生型相比，两种工程化mAb在高浓度下具有降低的粘度、较小的负第二维里系数和改善的溶解度。与建议的简单的设计原则得到的结果强调溶剂化效应的理解，并最终优化，高浓度的单克隆抗体溶液的性质的相关性，也可能对其他生物分子系统的影响。
Monoclonal antibody (mAb)-based therapeutics often require high-concentration formulations. Unfortunately, highly concentrated antibody solutions often have biophysical properties that are disadvantageous for therapeutic development, such as high viscosity, solubility limitations, precipitation issues, or liquid liquid phase separation. In this work, we present a computational rational design principle for improving the thermodynamic stability of mAb solutions through targeted point mutations. Two publicly available IgG1 monoclonal antibodies that exhibit high viscosity at high concentrations were used as model systems. Guided by a computationally efficient approach that combines molecular dynamics simulations with three-dimensional reference interaction site model theory, point mutations of charged residues were introduced in the variable Fv regions in such a manner that the hydration free energy was optimized. Two selected point mutants were then produced by transient expression and characterized experimentally. Both engineered mAbs have reduced viscosity at high concentration, less negative second virial coefficient, and improved solubility compared to the respective wild-types. The results obtained with the suggested straightforward design principle underline the relevance of solvation effects for understanding, and ultimately optimizing, the properties of highly concentrated mAb solutions, with possible implications also for other biomolecular systems.