The nonglycosylated variant in therapeutic monoclonal antibodies preferentially forms large aggregates under typical thermal stresses used in forced degradation studies.

IF 7.3 2区 医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL
mAbs Pub Date : 2025-12-01 Epub Date: 2025-08-10 DOI:10.1080/19420862.2025.2543768
Gao-Yuan Liu, Jenny Kim Kim, Shuli Tang, Yuetian Yan, Mandi Hopkins, Dalia Laredo, Teng-Chieh Yang, James Mutino, Douglas E Kamen, Kenneth S Graham, Mohammed Shameem, Shunhai Wang, Ning Li
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引用次数: 0

Abstract

Monoclonal antibodies (mAbs) feature a conserved N-linked glycosylation site in the CH2 domain, which exhibits heterogeneities in both occupancy and glycan structures. Previous studies have suggested that the unoccupied (nonglycosylated) variant exhibits decreased thermal stability, potentially impacting the overall stability of mAb products. This hypothesis, however, has remained largely unconfirmed, due to the low abundance of nonglycosylated variants in typical mAb products and the lack of effective analytical tools for detailed characterization of large aggregates with glycoform-specific information. Here, we used a postcolumn denaturation-assisted size exclusion chromatography mass spectrometry technique (SEC-PCD-MS) to reevaluate the effects of the nonglycosylated mAb variant on the thermal stability of mAb drugs during forced degradation studies. Our findings confirmed the compromised thermal stability of the nonglycosylated variant and its increased propensity to form large aggregates at elevated temperatures relevant to mAb-forced degradation studies. We also showed that this thermal stress-induced, nonglycosylation-mediated aggregation pathway could be widely observed in a diverse group of mAb molecules with varying properties. This study offers valuable insights into the rationale of selecting the appropriate temperature for mAb-forced degradation studies and highlights key considerations for data interpretation.

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治疗性单克隆抗体的非糖基化变体在强制降解研究中使用的典型热应力下优先形成大聚集体。
单克隆抗体(mab)在CH2结构域具有一个保守的n -链糖基化位点,其占有和糖基结构均具有异质性。先前的研究表明,未占据(非糖基化)的变体表现出较低的热稳定性,可能影响单抗产品的整体稳定性。然而,这一假设在很大程度上仍未得到证实,因为在典型的单抗产品中,非糖基化变体的丰度很低,而且缺乏有效的分析工具来详细表征具有糖型特异性信息的大聚集体。在这里,我们使用柱后变性辅助尺寸排除色谱-质谱技术(SEC-PCD-MS)重新评估非糖基化mAb变体在强制降解研究中对mAb药物热稳定性的影响。我们的研究结果证实了非糖基化变体的热稳定性受到损害,并且在与单克隆抗体强制降解研究相关的高温下形成大聚集体的倾向增加。我们还发现,这种热应力诱导的、非糖基化介导的聚集途径可以在不同性质的mAb分子中广泛观察到。这项研究为选择合适温度进行单克隆抗体强制降解研究的基本原理提供了有价值的见解,并强调了数据解释的关键考虑因素。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
mAbs
mAbs 工程技术-仪器仪表
CiteScore
10.70
自引率
11.30%
发文量
77
审稿时长
6-12 weeks
期刊介绍: mAbs is a multi-disciplinary journal dedicated to the art and science of antibody research and development. The journal has a strong scientific and medical focus, but also strives to serve a broader readership. The articles are thus of interest to scientists, clinical researchers, and physicians, as well as the wider mAb community, including our readers involved in technology transfer, legal issues, investment, strategic planning and the regulation of therapeutics.
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