Yüce Meral, Sert Fatma, Torabfam Milad, Parlar Ayhan, Gürel Büşra, Çakır Nilüfer, Dağlıkoca Duygu E, Khan Mansoor A, Çapan Yılmaz
Sabanci University, SUNUM Nanotechnology Research and Application Center, 34956, Istanbul, Turkey.
Sabanci University, Faculty of Engineering and Natural Sciences, 34956, Istanbul, Turkey; ILKO ARGEM Biotechnology R&D Center, 34906, Pendik, Istanbul, Turkey.
Anal Chim Acta. 2021 Apr 1;1152:238189. doi: 10.1016/j.aca.2020.12.064. Epub 2021 Jan 12.
The similarity between originator and biosimilar monoclonal antibody candidates are rigorously assessed based on primary, secondary, tertiary, quaternary structures, and biological functions. Minor differences in such parameters may alter target-binding, potency, efficacy, or half-life of the molecule. The charge heterogeneity analysis is a prerequisite for all biotherapeutics. Monoclonal antibodies are prone to enzymatic or non-enzymatic structural modifications during or after the production processes, leading to the formation of fragments or aggregates, various glycoforms, oxidized, deamidated, and other degraded residues, reduced Fab region binding activity or altered FcR binding activity. Therefore, the charge variant profiles of the monoclonal antibodies must be regularly and thoroughly evaluated. Comparative structural and functional analysis of physically separated or fractioned charged variants of monoclonal antibodies has gained significant attention in the last few years. The fraction-based charge variant analysis has proved very useful for the biosimilar candidates comprising of unexpected charge isoforms. In this report, the key methods for the physical separation of monoclonal antibody charge variants, structural and functional analyses by liquid chromatography-mass spectrometry, and surface plasmon resonance techniques were reviewed.
基于一级、二级、三级、四级结构和生物学功能，对原创单克隆抗体候选药物与生物类似药单克隆抗体候选药物之间的相似性进行严格评估。这些参数中的微小差异可能会改变分子的靶点结合、效力、疗效或半衰期。电荷异质性分析是所有生物治疗药物的先决条件。单克隆抗体在生产过程中或生产后容易发生酶促或非酶促结构修饰，导致片段或聚集体的形成、各种糖型、氧化、脱酰胺以及其他降解残基，降低Fab区域结合活性或改变FcR结合活性。因此，必须定期且全面地评估单克隆抗体的电荷变体图谱。在过去几年中，对单克隆抗体物理分离或分级的电荷变体进行比较结构和功能分析受到了广泛关注。基于分级的电荷变体分析已被证明对包含意外电荷异构体的生物类似药候选药物非常有用。在本报告中，综述了单克隆抗体电荷变体物理分离、液相色谱 - 质谱联用和表面等离子体共振技术进行结构和功能分析的关键方法。
