Waters Laura J, Whiteley Joseph, Small William, Mellor Steve
School of Applied Sciences, University of Huddersfield, Queensgate, Huddersfield, HD1 3DH, UK.
Croda Europe Ltd, Cowick Hall, Snaith, Goole, DN14 9AA, UK.
Heliyon. 2023 Oct 26;9(11):e21712. doi: 10.1016/j.heliyon.2023.e21712. eCollection 2023 Nov.
Protein stability is fundamental to maintain pharmaceutical efficacy in the nascent field of biologics. One particular property that is essential for therapeutic effect is retention of the folded 3-dimensional conformation, i.e. once unfolding has occurred the biologic is often rendered inactive. In this work we propose a modified form of a recently published UV spectroscopic method that identifies protein unfolding. In this study we determine concentration limits to avoid protein unfolding of two model surfactants, namely polysorbate 20 and polysorbate 80, by correlating surfactant concentration with percentage 'unfolded' for three model proteins. For each scenario two distinct regions were observed, firstly surfactant concentrations at which no unfolding had occurred, followed by a second region whereby unfolding steadily increased with surfactant concentration. In general for the combinations analysed in this study, this second region began to appear around ten times below the critical micellar concentration of each surfactant, regardless of the protein or polysorbate chosen. It is therefore proposed that this adapted method could be used by researchers in the early stages of formulation development as a convenient and simple screening tool to confirm the 'onset of unfolding' concentration for protein-surfactant formulations, thus helping to optimise surfactant concentration selection in pharmaceutical formulations to maintain the benefits of surfactants yet avoid inadvertent unfolding.
在生物制品这个新兴领域，蛋白质稳定性对于维持药物疗效至关重要。对于治疗效果而言，一个特别重要的特性是保持折叠的三维构象，也就是说，一旦发生解折叠，生物制品通常就会失去活性。在这项工作中，我们提出了一种对最近发表的用于识别蛋白质解折叠的紫外光谱方法的改进形式。在本研究中，我们通过将三种模型蛋白质的表面活性剂浓度与“解折叠”百分比相关联，来确定两种模型表面活性剂（即聚山梨酯20和聚山梨酯80）避免蛋白质解折叠的浓度极限。对于每种情况，都观察到两个不同的区域，首先是未发生解折叠的表面活性剂浓度区域，其次是解折叠随表面活性剂浓度稳步增加的第二个区域。一般来说，对于本研究中分析的组合，无论选择何种蛋白质或聚山梨酯，这个第二个区域开始出现的浓度大约是每种表面活性剂临界胶束浓度的十分之一。因此，建议研究人员在制剂开发的早期阶段可以使用这种改进的方法，作为一种方便简单的筛选工具，来确定蛋白质 - 表面活性剂制剂的“解折叠起始”浓度，从而有助于优化药物制剂中表面活性剂浓度的选择，在保持表面活性剂益处的同时避免意外的解折叠。
