Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal.
Novartis Pharma AG, 4056 Basel, Switzerland.
J Pharm Biomed Anal. 2021 Jan 5;192:113650. doi: 10.1016/j.jpba.2020.113650. Epub 2020 Sep 25.
Identification of molecular liabilities and implementation of mitigation strategies are key aspects that need to be considered by pharmaceutical companies developing therapeutic proteins. In the field of monoclonal antibodies, an efficient and streamlined process known as developability assessment is used for the selection of the "fittest" candidate. Other protein modalities, have in most cases only a limited number of possible candidates, requiring a paradigm change to a concept of candidate enabling. The assessment of liabilities at early project phases with the possibility to re-engineer candidates becomes essential for the success of these projects. Each protein possesses a unique stability profile resulting from the interplay of conformational, colloidal, chemical and physical stability attributes. All of these attributes strongly depend on external factors. Conformational and colloidal stability profiles of three non-immunoglobulin domain based proteins, namely Carbonic anhydrase, Ovalbumin and Thyroglobulin, and of two monoclonal antibodies were assessed in dependence of solution pH, ionic strength and varying buffering agents. The impact of screened external factors on proteins' stability attributes varied significantly, indicating presence of molecule specific liabilities. Screening of such a broad space of conditions at early project phases is only feasible using low-material consuming, high-throughput analytical methods as exemplified in this study.
鉴定分子缺陷并实施缓解策略是制药公司开发治疗性蛋白时需要考虑的关键方面。在单克隆抗体领域，有一种称为可开发性评估的高效简化流程，用于选择“最合适”的候选物。其他蛋白质形式在大多数情况下只有数量有限的可能候选物，需要将概念从候选物选择转变为候选物设计。在早期项目阶段评估缺陷，并有可能对候选物进行重新设计，这对于这些项目的成功至关重要。每种蛋白质都具有独特的稳定性特征，这是由构象、胶体、化学和物理稳定性属性相互作用的结果。所有这些属性都强烈依赖于外部因素。本文评估了三种非免疫球蛋白结构域蛋白（即碳酸酐酶、卵清蛋白和甲状腺球蛋白）和两种单克隆抗体的构象和胶体稳定性特征，这些蛋白的稳定性特征取决于溶液 pH 值、离子强度和不同的缓冲剂。筛选出的外部因素对蛋白质稳定性属性的影响差异很大，表明存在分子特异性缺陷。只有使用低材料消耗、高通量的分析方法（如本研究所示），才能在早期项目阶段筛选如此广泛的条件。
