Department of Biochemical Engineering, University College London, Gower Street, London WC1E 6BT, United Kingdom.
National Measurement Laboratory at LGC Ltd., Queens Road, Teddington TW11 0LY, United Kingdom.
Mol Pharm. 2020 Dec 7;17(12):4637-4651. doi: 10.1021/acs.molpharmaceut.0c00877. Epub 2020 Oct 28.
Assuring the stability of therapeutic proteins is a major challenge in the biopharmaceutical industry, and a better molecular understanding of the mechanisms through which formulations influence their stability is an ongoing priority. While the preferential exclusion effects of excipients are well known, the additional presence and impact of specific protein-excipient interactions have proven to be more elusive to identify and characterize. We have taken a combined approach of in silico molecular docking and hydrogen deuterium exchange-mass spectrometry (HDX-MS) to characterize the interactions between granulocyte colony-stimulating factor (G-CSF), and some common excipients. These interactions were related to their influence on the thermal-melting temperatures () for the nonreversible unfolding of G-CSF in liquid formulations. The residue-level interaction sites predicted in silico correlated well with those identified experimentally and highlighted the potential impact of specific excipient interactions on the of G-CSF.
确保治疗性蛋白质的稳定性是生物制药行业面临的主要挑战，更好地理解制剂影响其稳定性的机制是当前的重点。虽然赋形剂的优先排除效应是众所周知的，但特定蛋白质-赋形剂相互作用的额外存在和影响更难以确定和表征。我们采用了计算机分子对接和氘氢交换-质谱（HDX-MS）相结合的方法，来描述粒细胞集落刺激因子（G-CSF）与一些常见赋形剂之间的相互作用。这些相互作用与它们对 G-CSF 在液体制剂中不可逆展开的热熔融温度（）的影响有关。计算机预测的残基水平相互作用位点与实验中鉴定的相互作用位点很好地相关，突出了特定赋形剂相互作用对 G-CSF 的影响。
