化学
圆二色性
阳离子聚合
动态光散射
离子强度
化学稳定性
差示扫描量热法
色谱法
异构化
分子
蛋白质聚集
单克隆抗体
生物物理学
立体化学
结晶学
有机化学
生物化学
催化作用
抗体
化学工程
水溶液
物理
生物
纳米颗粒
工程类
热力学
免疫学
作者
Tingting Wang,Ozan S. Kumru,Yi Li,Y. John Wang,Jennifer Zhang,Jae Hyun Kim,Sangeeta B. Joshi,C. Russell Middaugh,David B. Volkin
摘要
ABSTRACT
Monoclonal antibody (mAb) fragments are emerging as promising alternatives to full-length mAbs as protein therapeutic candidates. Antigen-binding fragments (Fabs) are the most advanced with three Fab-based drug products currently approved. This work presents preformulation characterization data on the effect of pH, NaCl concentration, and various cationic excipients on the physical and chemical stability of a Fab molecule with multiple negatively charged Asp residues in the complementarity-determining region. Conformational stability was evaluated using an empirical phase diagram approach based on circular dichroism, intrinsic Trp and extrinsic 8-anilino-1-naphthalene sulfonate (ANS) fluorescence, and static light scattering measurements. The effect of NaCl concentration, various cationic excipients and pH on the Fab molecule's conformational stability, aggregation propensity, and chemical stability (Asp isomerization) was determined by differential scanning calorimetry, optical density measurements at 350nm (OD350), and ion-exchange chromatography, respectively. Increasing NaCl concentration increased the overall conformational stability, decreased aggregation rates, and lowered the rates of Asp isomerization. No such trends were noted for pH or cationic excipients. The potential interrelationships between protein conformational and chemical stability are discussed in the context of designing stable protein formulations.
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