Influence of histidine on the stability and physical properties of a fully human antibody in aqueous and solid forms
Influence of histidine on the stability and physical properties of a fully human antibody in aqueous and solid forms
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DOI：
10.1023/b:pham.0000008042.15988.c0
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发表时间：
2003-12-01
影响因子：
3.7
通讯作者：
Chamow, SM
中科院分区：
文献类型：
--
作者：
Chen, B;Bautista, R;Chamow, SM
目的。本研究的目的是研究组氨酸对水性和固体形式的全人抗 IL8 单克隆抗体 (ABX-IL8) 的稳定性和物理性质的影响。方法。使用部分因子设计，我们测试了许多赋形剂，包括组氨酸、蔗糖和其他常用赋形剂，以了解液体和冻干形式抗体的稳定性和物理性质。使用尺寸排阻高效液相色谱（SEC-HPLC）、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）和粘度计评估抗体稳定性和物理性质。残余水分含量通过卡尔费休库仑滴定仪测定。使用差示扫描量热法 (DSC) 检测固体块的玻璃化转变温度 (T-g) 和液体制剂中抗体的熔化温度 (T-m)。使用傅里叶变换红外（FTIR）光谱来检查整体二级结构。结果。增加本体溶液中的组氨酸浓度可抑制冻干和储存时高分子量 (HMW) 物质和聚集体的增加。此外，大量组氨酸增强了抗体在冷冻和热应激条件下的溶液稳定性，聚集体水平较低就证明了这一点。此外，组氨酸降低了抗体溶液的粘度，这对于剂型的制造是理想的。然而，在批量冻融过程中将制剂暴露于不锈钢容器后，液体制剂中的高浓度组氨酸导致溶液着色，并且在高温（40℃）储存时出现高水平的聚集体。结论。组氨酸增强了 ABX-IL8 水剂和冻干剂的稳定性。组氨酸还可以改善物理性质，例如降低溶液粘度。含有高浓度组氨酸的液体制剂不应在高温下储存在不锈钢罐中。
Purpose. The aim of the study was to investigate the effect of histidine on the stability and physical properties of a fully human anti-IL8 monoclonal antibody (ABX-IL8) in aqueous and solid forms.Methods. Using a fractional factorial design, we tested many excipients, including histidine, sucrose, and other commonly used excipients, on the stability and physical properties of the antibody in both liquid and lyophilized forms. Antibody stability and physical properties were evaluated using size-exclusion high-performance liquid chromatography (SEC-HPLC), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and a viscometer. Residual moisture content was determined by coulometric Karl Fischer titrator. Differential scanning calorimetry (DSC) was used to detect the glass transition temperatures (T-g) of the solid cakes and melting temperatures (T-m) of the antibody in liquid formulations. Fourier-transform infrared (FTIR) spectroscopy was used to examine the overall secondary structure.Results. Increasing the histidine concentration in the bulk solution inhibited the increases of high-molecular-weight (HMW) species and aggregates upon lyophilization and storage. In addition, histidine bulk enhanced solution stability of the antibody under freezing and thermal stress conditions, as evidenced by the lower levels of aggregates. Furthermore, histidine reduced viscosity of the antibody solution, which is desirable for the manufacture of the dosage form. However, high concentrations of histidine in liquid formulations led to coloration of the solution and high levels of aggregates on storage at elevated temperature (40 degreesC) after the formulations were exposed to stainless steel containers during bulk freezing-thawing.Conclusions. Histidine enhanced the stability of ABX-IL8 in both aqueous and lyophilized forms. Histidine also improved the physical properties such as reducing the solution viscosity. Liquid formulations containing high concentrations of histidine should not be stored in stainless steel tanks at elevated temperatures.