Analytical Development Biologicals, Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach an der Riss, Germany.
Institute of Pharmaceutical Biotechnology, Ulm University, Ulm, Germany.
MAbs. 2024 Jan-Dec;16(1):2400414. doi: 10.1080/19420862.2024.2400414. Epub 2024 Sep 8.
During the development process of therapeutic monoclonal antibodies (mAbs), it is crucial to control (critical) quality attributes such as N-glycosylation influencing pharmacokinetics (PK) and Fc effector functions. Previous reports have shown that mAbs containing high-mannose N-glycans are cleared faster from blood circulation, leading to reduced half-lives. The high-mannose N-glycan content of mAbs can be influenced during the cell culture process by factors such as cell lines, process conditions, and media. Furthermore, mAbs have either one high mannose N-glycan (asymmetrical high-mannose glyco-pair) or two high mannose N-glycans (symmetrical high-mannose glyco-pair). The hypothesis that the mannose receptor (MR, CD206) accelerates clearance by facilitating their internalization and subsequent lysosomal degradation is widespread. However, the interaction between MR and mAbs has not been explicitly demonstrated. This study aimed to investigate this interaction, providing the first systematic demonstration of MR binding to the Fc region of mAbs with high-mannose N-glycans. Two novel analytical methods, MR surface plasmon resonance and MR affinity chromatography, were developed and applied to investigate the MR-mAb interaction. The interaction is found to be dependent on high-mannose content, but is independent of the mAb format or sequence. However, different glyco-pairs exhibited varying binding affinities to the MR, with the symmetrical high-mannose glyco-pair showing the strongest binding properties. These findings strengthen the hypothesis for the MR-mediated mAb interaction and contribute to a deeper understanding of the MR-mAb interaction, which could affect the criticality of high-mannose containing mAbs development strategies of IgG-based molecules and improve their PK profiles.
在治疗性单克隆抗体 (mAb) 的开发过程中，控制影响药代动力学 (PK) 和 Fc 效应功能的关键质量属性（如 N-糖基化）至关重要。先前的报告表明，含有高甘露糖 N-糖的 mAb 从血液循环中更快地清除，导致半衰期缩短。mAb 的高甘露糖 N-糖含量可通过细胞系、工艺条件和培养基等因素在细胞培养过程中受到影响。此外，mAb 要么含有一个高甘露糖 N-糖（不对称高甘露糖糖对），要么含有两个高甘露糖 N-糖（对称高甘露糖糖对）。甘露糖受体 (MR, CD206) 通过促进其内化和随后的溶酶体降解来加速清除的假设已被广泛接受。然而，MR 与 mAb 之间的相互作用尚未得到明确证明。本研究旨在研究这种相互作用，首次系统地证明了 MR 与含有高甘露糖 N-糖的 mAb 的 Fc 区域结合。开发了两种新的分析方法，即 MR 表面等离子体共振和 MR 亲和层析，并应用于研究 MR-mAb 相互作用。结果表明，这种相互作用依赖于高甘露糖含量，但与 mAb 结构或序列无关。然而，不同的糖对与 MR 的结合亲和力不同，对称高甘露糖糖对表现出最强的结合特性。这些发现支持了 MR 介导的 mAb 相互作用的假设，并有助于更深入地了解 MR-mAb 相互作用，这可能会影响基于 IgG 分子的高甘露糖含量 mAb 开发策略的关键性，并改善其 PK 特征。
