Impact of dextran on thermal properties, product quality attributes, and monoclonal antibody stability in freeze-dried formulations
Impact of dextran on thermal properties, product quality attributes, and monoclonal antibody stability in freeze-dried formulations
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DOI：
10.1016/j.ejpb.2019.12.010
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发表时间：
2020-02-01
影响因子：
4.9
通讯作者：
Allmendinger, Andrea
中科院分区：
文献类型：
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作者：
Haeuser, Christina;Goldbach, Pierre;Allmendinger, Andrea
冷冻干燥通常用于改善不稳定生物药物的液体制剂的稳定性。冻干保护剂和冷冻保护剂如蔗糖传统上用作赋形剂，但具有低的玻璃化转变温度（T-g ′）和塌陷温度（T-c）。因此，这些制剂需要低的初级干燥温度，使得冻干循环耗时且昂贵。我们研究了不同的葡聚糖（1，40，150，和500 kDa）和混合物的葡聚糖与蔗糖作为替代赋形剂。以牛血清白蛋白为模型蛋白，研究了右旋糖酐对固体状态下热性质、饼外观和其他质量属性的影响。特别是在葡聚糖与蔗糖的较高重量比下，40 - 500 kDa的中至高分子量（M-w）的葡聚糖显示出比蔗糖高20 ℃的T-c，这反映在优雅的冻干物中。然而，这导致复溶时间较慢。与蔗糖相比，葡聚糖的添加导致较低的残留水分水平和较高的T-g值。我们确认了两种单克隆抗体（mAb）在具有不同M-w的蔗糖和葡聚糖的两种重量比下的热性质，并测试了在40 ° C下14天的稳定性。虽然储存后未观察到抗体相对效价的损失，但尺寸排阻色谱和等电聚焦显示高分子量物质（HMW）和酸性物质的强烈增加，这取决于葡聚糖的Mw。通过硼酸盐亲和色谱和质谱分析对所选制剂（右旋糖酐1 kDa）进行进一步表征，我们证明HMW是右旋糖酐游离末端葡萄糖糖化的结果。当加入蔗糖时，这种化学修饰强烈减少，蔗糖可能通过屏蔽其表面来保护蛋白质。我们的研究结果表明，制剂科学家需要谨慎使用右旋糖酐作为冻干mAb制剂的辅料。然而，适当比例的蔗糖和葡聚糖的二元混合物可能优于纯蔗糖制剂，允许更快的冷冻干燥循环，从而产生优雅的冻干物和良好的蛋白质稳定性。
Freeze-drying is commonly used to improve stability of liquid formulations of labile biopharmaceuticals. Lyo- and cryoprotectants such as sucrose are traditionally utilized as excipients, but have low glass transition (T-g') and collapse temperatures (T-c), Consequently, these formulations require low primary drying temperatures making the lyophilization cycle time-consuming and costly. We investigated different dextrans (1, 40, 150, and 500 kDa) and mixtures of dextran with sucrose as alternative excipients. The influence of dextran on thermal properties, cake appearance, and other quality attributes in the solid state was studied using bovine serum albumin as model protein. Especially at higher weight ratios of dextran to sucrose, dextrans of medium to high molecular weight (M-w) of 40-500 kDa showed up to 20 degrees C higher T-c compared to sucrose, which was reflected in elegant lyophilisates. However, this resulted in slower reconstitution times. Addition of dextran led to lower residual moisture levels and higher T-g values compared to sucrose. We confirmed the thermal properties for two monoclonal antibodies (mAb) at two weight ratios of sucrose and dextran with different M-w, and tested for stability at 40 degrees C for 14 days. While no loss in relative potency of the antibodies was observed after storage, size exclusion chromatography and isoelectric focusing revealed a strong increase in high molecular weight species (HMWs) and acidic species, which were dependent on the M w of the dextrans. With further characterization of selected formulations (dextran 1 kDa) by boronate affinity chromatography and mass spectrometry analysis, we demonstrated that HMWs were a result of glycation by free terminal glucose of the dextran. This chemical modification was strongly reduced when adding sucrose, which protects the protein possibly by shielding its surface. Our results demonstrate that formulation scientists need to use dextrans as excipients in freeze-dried mAb formulations with caution. A binary mixture of sucrose and dextran in adequate ratio however might potentially be superior to pure sucrose formulations allowing for faster freeze-drying cycles resulting in elegant lyophilisates and good protein stability.