单克隆抗体药物中微量蛋白质含量测定方法的建立  

Establishment of method for determining trace protein content in monoclonal antibody drugs

作　　者：崔春博 武刚[1] 俞小娟 刘春雨[1] 李萌[1] 倪永波 崔永霏 于传飞[1] 王兰[1] CUI Chunbo;WU Gang;YU Xiaojuan;LIU Chunyu;LI Meng;NI Yongbo;CUI Yongfei;YU Chuanfei;WANG Lan(Division of Monoclonal Antibody Products,National Institutes for Food and Drug Control,Key Laboratory of the Ministry of Health for Research on Quality and Standardization of Biotech Products,Key Laboratory of the National Medical Products Administration for Research on Quality Study and Evaluation of Biological Products,Products,Beijing 102629,China)

机构地区：[1]中国食品药品检定研究院单克隆抗体产品室,卫生部生物技术产品检定及标准化重点实验室,国家药品监督管理局生物制品质量研究与评价重点实验室,北京102629

出　　处：《山西医科大学学报》2025年第3期290-293,共4页Journal of Shanxi Medical University

基　　金：国家科技重大专项“重大新药创制”项目(2019ZX09732-002);中山市重大科技专项-战略性新兴产业技术公关专题(210204163866513)。

摘　　要：目的建立并验证一种测定单克隆抗体药物中微量蛋白质含量的方法。方法采用高效液相色谱-荧光检测器(high performance liquid chromatography-fluorescence detector,HPLC-FLD)法建立单抗药物中蛋白质含量的检测方法,并对方法的专属性、线性、重复性、中间精密度、准确度和耐用性进行验证。结果该方法专属性强。在250~750 ng进样量范围内,蛋白质进样量与峰面积呈良好的线性关系,决定系数R^(2)>0.99。两名实验人员分别在3个工作日对同一份供试品溶液进行18次独立测定,其蛋白质含量的变异系数为3.03%,蛋白质含量验证溶液(550 ng)的回收率在90%~110%内。6次蛋白质含量验证溶液(550 ng)蛋白质含量测定值的RSD为2.88%。不同色谱柱检测蛋白质含量验证溶液(550 ng)的变异系数为1.86%。结论成功建立了检测单克隆抗体药物中微量蛋白质含量的高效液相色谱法,该方法专属性强、重复性好、精密度及准确度高,可对单克隆抗体药物中微量蛋白质含量进行质量控制。Objective To establish and validate a method for the detection of trace protein content in monoclonal antibody drugs.Methods The detection method for protein content in monoclonal antibody drugs was established by high performance liquid chromatography-fluorescence detector(HPLC-FLD)method,and the specificity,linearity,repeatability,intermediate precision,accuracy and robustness of the method were validated.Results The specificity of the method was proved to be good.The injection volume and the peak area of proteins showed a good linear relationship in the range of 250-750 ng injection volume,with a coefficient of determination R^(2)>0.99.The coefficients of variation for the 18 independent tests of the same sample by two staffs over three days was 3.03%,and the recovery rates of the protein content validation solutions(550 ng)were in the range of 90%-110%.The RSD of measured value of validation solution at 550 ng injection volume was 2.88%after six times injection.The coefficient of variation of the validation solution(550 ng)for protein content was 1.86%for different columns.Conclusion A HPLC method for trace protein content determination in monoclonal antibody drugs is successfully developed with good repeatability,specificity,precision and accuracy,which can be used for quality control of trace protein contents in monoclonal antibody drugs.

关 键 词：高效液相色谱 荧光检测器 单克隆抗体药物 微量 含量测定 质量控制 

分 类 号：R737.25[医药卫生—肿瘤]