玻璃内包材界面修饰对机械应力诱导的单克隆抗体聚集体形成的影响

Effect of glass primary container surface modification on monoclonal antibody aggregates induced by mechanical stress

利用十八烷基三氯硅烷(octadecyltrichlorosilane,OTS)对中硼硅玻璃管制注射剂瓶内表面进行化学改性,使用接触角仪(水接触角从50°±1°增加至90°±2°)、原子力显微镜(表面粗糙度从0.448±0.086增加至1.282±0.117)、红外(出现—CH_(2)—和—CH_(3)特征峰)对表面进行验证,成功制备了具有疏水性能的玻璃表面。给玻璃瓶中的单克隆抗体施加机械应力,使用微流成像技术、尺寸排阻-高效液相色谱法(size exclusion-high performance liquid chromatography,SE-HPLC)、外源荧光对单抗制剂的稳定性进行全面表征。结果表明,OTS处理的疏水性界面可以减少机械应力诱导的蛋白质聚集体的产生。由于疏水表面与蛋白质强的疏水相互作用,OTS处理的疏水表面可有效抑制机械应力诱导的抗体分子的聚集。

The inner surface of middle borosilicate glass tubing was modified by octadecyltrichlorosilane(OTS).The contact angles of modified surfaces which were measured by contact angle goniometer increased from 50°±1°to 90°±2°.The surface roughness which was measured by atomic force microscope increased from 0.448±0.086 to 1.282±0.117,and the result of Fourier transform infrared spectroscopy(FTIR)indicated that the OTS was successfully coated on the glass surfaces.The monoclonal antibody in glass containers were subjected to mechanical stress.Microflow imaging,size exclusion-high performance liquid chromatography(SE-HPLC)and extrinsic fluorescence dye were used to characterize the stability of monoclonal antibody.The results showed that the OTS-treated hydrophobic interface could reduce the generation of mechanical stress-induced protein aggregates.Due to the strong hydrophobic interaction between hydrophobic surface and protein,the hydrophobic surface could effectively reduce the aggregation of antibody molecules induced by mechanical stress.