尼妥珠单抗毛细管区带电泳鉴别方法的建立与验证

Establishment and validation of nimotuzumab identification method with capillary zone electrophoresis

目的:建立尼妥珠单抗的毛细管区带电泳鉴别分析方法。方法:利用基于荷质比的毛细管区带电泳技术建立尼妥珠单抗的鉴别分析方法,并对所建立的方法进行系统适用性、专属性、精密度等方法学验证。结果:利用该方法对各单抗参比品进行测定,系统适用性中参比品的主峰迁移时间差均<0.1 min,该方法能够对尼妥珠单抗和其他单抗进行有效的区分,不同单抗的主峰迁移时间差均>0.1 min,在毛细管区带电泳色谱行为上显示出产品专属性。在日内精密度试验中,重复制备样品及重复进样的主峰迁移时间RSD值分别为0.081%和0.035%;在日间精密性评价中,主峰迁移时间的RSD值为0.566%。结论:建立了简便、快速、准确的尼妥珠单抗毛细管区带电泳鉴别方法,可用于尼妥珠单抗及其他抗体类产品的常规检定。

Objective： To develop the identification method for nimotuzumab using capillary zone electrophoresis（ CZE）. Methods： Firstly,the identification method of nimotuzumab was established using capillary zone electrophoresis. Then,its system suitability,specificity and precision were validated. Results： Other 8 monoclonal antibodies（ mAbs） as the physicochemical reference were determined using the established CZE method. For the system suitability study,all the difference of main peak migration time between the repeated reference injections was smaller than 0. 1 min. For specificity study,the CZE method could effectively distinguish nimotuzumab from other mAbs,and all the difference of main peak migration time between these samples was greater than 0. 1 min,which showed the product specificity on chromatographic behavior of CZE method. For intra-day precision,the RSD of the main peak migration time for repeated preparation samples and repeated injection samples was 0. 081%and 0. 035%,respectively. For inter-day precision,the RSD of the main peak migration time was 0. 566%. These results showed that the established CZE method had good precision. Conclusion： The identification of nimotuzumab analyzed by the CZE method developed in this study is simple,fast and accurate. Thus,it could offer a promising method for quality control and evaluation of nimotuzumab and other antibody-based biotherapeutics.