首批HPLC-SEC系统适用性免疫球蛋白国家对照品研制

目的 研制一种高效液相分子排阻色谱(HPLC-SEC)系统适用性免疫球蛋白国家对照品,用于免疫球蛋白类产品等分子大小分布系统适用性确认。方法 制备符合技术指标要求的静注人免疫球蛋白产品作为对照品原料,合并后进行除菌过滤,最后分装。按照《中国药典》2020年版三部附录中“人免疫球蛋白类制品IgG单体加二聚体测定方法”,在8家实验室进行协作标定,同时进行稳定性监测。结果 8家实验室测定本批对照品单体与裂解物分离度(R)均＞1.5(均值为2.3),实验条件均符合《中国药典》2020年版三部中相关要求;8家实验室对照品裂解物峰面积百分比均值为3.83%,95%置信区间(3.48,4.19)%,均＞2.0%,满足预期技术要求。在2~8 ℃下持续对该对照品进行14个月稳定性监测,分离度(R)符合《中国药典》要求,裂解物峰面积百分比未发生趋势性变化,建议对照品按2~8 ℃保存和运输。结论 HPLC-SEC系统适用性免疫球蛋白国家对照品可以用于《中国药典》2020年版要求的系统适用性测试,有助于提升免疫球蛋白等产品分子大小分布检测结果可靠性。

Development of the First Batch of National Reference Substance of Immunoglobulin for HPLC-SEC System Suitability Test

OBJECTIVE To establish a national reference substance of immunoglobulin for HPLC-SEC system suitability test to confirm the system suitability of HPLC molecular size distribution analytical assay of immunoglobulin products. METHODS Intravenous human immunoglobulin products meeting the technical requirements were prepared as the raw materials of the reference substance, which were treated, combined, sterilized and filtered, and then sub packaged. According to the 2020 edition of Chinese Pharmacopoeia part III human immunoglobulin IgG monomer and dimer determination method, eight laboratories were involved for collaborative calibration. Stability monitoring was also carried out meanwhile. RESULTS The mean value of resolution of monomer vs fragments from the eight laboratories was 2.3, all of which were greater than 1.5, which verified that the experimental conditions met the requirements of Chinese Pharmacopoeia 2020 edition. The inter-lab mean value of peak area% of the national reference substance for immunoglobulin HPLC-SEC system suitability test was 3.83% (95% CI:3.48%-4.19%), all of which were greater than 2.0%, meeting the expected technical requirements. The stability of national reference substance for immunoglobulin HPLC-SEC system suitability test was continuously monitored at 2-8 ℃ for 14 m. The resolution data complied with the requirements of Chinese Pharmacopoeia (R＞1.5) and the peak area% of fragments data showed no abnormal trends. It was suggested that the reference substance be stored and transported at 2-8 ℃. CONCLUSIONT The national reference substance of immunoglobulin for HPLC-SEC system suitability test can achieve the system applicability test required by the Chinese Pharmacopoeia 2020 edition, which is helpful to improve the reliability of the molecular size distribution test results of immunoglobulin and other products.