Understanding the effects of disulfide bond reduction on the structure, function and stability of monoclonal antibodies used in therapy

最先进的生物治疗药物，如单克隆抗体（mab）含有二硫键。单克隆抗体是使用重组细胞表达系统制造的，该系统可以将硫醇还原酶分泌到培养基中。这些酶可以减少抗体中的二硫键，在制造过程中引起结构和功能的变化。此外，通常用这种抗体治疗的癌症和炎症性疾病等疾病也会分泌大量的还原酶，这意味着在给药后，这种抗体可以在体内被修饰。本项目的目的是表征二硫键还原对单克隆抗体配制后结构、功能和稳定性的影响。最先进的生物物理分析（质谱分析）和功能生物分析相结合，将阐明还原对结构和功能的影响。这将提供基本的结构知识，以提高生物治疗药物的安全性和有效性，并将为改进基于抗体的产品的制造过程和配方提供途径。这项工作与EPSRC制造未来的主题一致，并且是应对人口老龄化发展医疗保健挑战的战略契合。该项目涉及与国家生物标准与控制研究所（NIBSC）以及LGC的专业知识和合作，以补充伦敦大学学院生化工程系的分析，结构，计算和配方专业知识。

State of the art biotherapeutics such as monoclonal antibodies (mAbs) contain disulfide bonds. MAbs are manufactured using recombinant cellular expression systems, which can secrete thiol reductase enzymes into the culture medium. These enzymes can reduce disulfide bonds in the antibodies, causing structural and functional changes during manufacturing. Furthermore, diseases such as cancer and inflammatory disorders that are commonly treated by such antibodies also secrete large amounts of reductases, meaning the antibodies can be modified in vivo after administration of the formulated drug. The aim of this project is to characterise the effects of disulfide bond reduction on the structure, function and stability after formulation of mAbs. A combination of state-of-the-art biophysical analysis by mass spectrometry, and bioassays for function, will elucidate the impact of reduction on both structure and function. This will provide fundamental structural knowledge to improve safety and efficacy of biotherapeutics in general and will inform routes to improve the manufacturing process and the formulation of antibody-based products. This work aligns to the EPSRC Manufacturing the Future theme, and is a strategic fit to the challenge of developing healthcare for an ageing population.The project involves expertise and collaboration with the National Institute of Biological Standards and Control (NIBSC), and also LGC, to complement the analytical, structural, computational and formulation expertise of the UCL Department of Biochemical Engineering.