Chen Bei, Bautista Raquel, Yu Kwok, Zapata Gerardo A, Mulkerrin Michael G, Chamow Steven M
Department of Process Sciences, Abgenix, Inc., Fremont, CA 94555, USA.
Pharm Res. 2003 Dec;20(12):1952-60. doi: 10.1023/b:pham.0000008042.15988.c0.
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.
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 (Tg) of the solid cakes and melting temperatures (Tm) of the antibody in liquid formulations. Fourier-transform infrared (FTIR) spectroscopy was used to examine the overall secondary structure.
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 degrees C) after the formulations were exposed to stainless steel containers during bulk freezing-thawing.
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.
本研究旨在探讨组氨酸对全人源抗IL8单克隆抗体（ABX - IL8）水溶液和固体形式稳定性及物理性质的影响。
采用部分因子设计，我们测试了多种辅料，包括组氨酸、蔗糖及其他常用辅料，对液体和冻干形式抗体的稳定性及物理性质的影响。使用尺寸排阻高效液相色谱法（SEC - HPLC）、十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳法（SDS - PAGE）和粘度计评估抗体的稳定性和物理性质。用库仑卡尔费休滴定仪测定残留水分含量。差示扫描量热法（DSC）用于检测固体饼剂的玻璃化转变温度（Tg）和液体制剂中抗体的熔点温度（Tm）。采用傅里叶变换红外（FTIR）光谱法检测整体二级结构。
增加本体溶液中组氨酸浓度可抑制冻干和储存过程中高分子量（HMW）物种和聚集体的增加。此外，组氨酸本体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，这对于剂型制造是有利的。然而，液体制剂中高浓度的组氨酸会导致溶液变色，并且在本体冻融过程中制剂暴露于不锈钢容器后，在高温（40℃）储存时会产生高水平的聚集体。
组氨酸增强了ABX - IL8水溶液和冻干形式的稳定性。组氨酸还改善了物理性质，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。
