Institute for Analytical and Bioanalytical Chemistry, University of Ulm, Ulm D-89069, Germany.
Boehringer Ingelheim Pharma GmbH & Co. KG, Analytical Development Biologicals, Biberach D-88397, Germany.
J Phys Chem B. 2024 Oct 24;128(42):10408-10416. doi: 10.1021/acs.jpcb.4c05358. Epub 2024 Oct 10.
We study the influence of urea on the stability of monoclonal antibodies (mAbs) using molecular dynamics (MD) simulations in combination with differential scanning fluorimetry (DSF). We show that a denaturing cosolute such as urea binds strongly to the protein, which can lead to denaturation and enhanced aggregation behavior at high temperatures. The interaction between protein and urea crucially depends on the surface properties of the individual mAb domains and therefore affects the general binding to the protein differently. The study of these mechanisms for proteins with multiple domains, such as mAbs, encounters significant limitations in experimental analysis methods due to their complexity. Using computational and experimental methods, we are able to separate the protein-urea interaction by domain and show that Lennard-Jones interactions are mainly responsible for significant binding effects. Our results emphasize the potential of MD simulations in combination with Kirkwood-Buff theory to study the interactions between proteins with multiple domains and cosolutes as formulation excipients for drug discovery and development.
我们使用分子动力学 (MD) 模拟结合差示扫描荧光法 (DSF) 研究了尿素对单克隆抗体 (mAb) 稳定性的影响。结果表明，变性共溶剂（如尿素）与蛋白质结合紧密，这可能导致在高温下变性和增强聚集行为。蛋白质与尿素之间的相互作用取决于单抗结构域的表面特性，因此会对整体与蛋白质的结合产生不同的影响。对于具有多个结构域的蛋白质（如 mAb），由于其复杂性，实验分析方法在研究这些机制方面存在显著的局限性。通过使用计算和实验方法，我们能够按结构域分离蛋白质-尿素相互作用，并表明伦纳德-琼斯相互作用主要负责显著的结合效应。我们的结果强调了 MD 模拟与 Kirkwood-Buff 理论相结合在研究具有多个结构域的蛋白质与共溶剂之间相互作用的潜力，这对于药物发现和开发中的配方赋形剂具有重要意义。
