Method Validation of Methodology Similarity between Rituximab and Its Biosimilar by ELISA
WANG Meng-jia1, DONG Li-hou2, LI Wan-wan3, LÜ Xiao-long3, WANG Bian-zhen3*, GE Zhi-qiang1*
1. School of Chemical Engineering and Technology, Tianjing University, Tianjin 300072, China; 2. Institute of Military Medicine, Academy of Military Sciences, Beijing 102206, China; 3. United-Power Pharma Tech Co., Ltd., Beijing 102206, China
Abstract：OBJECTIVE To validate the methodology similarity of the biosimilar (recombinant anti-human CD20 chimeric monoclonal antibody, B) and the reference (rituximab, R). METHODS The biosimilar and the reference were measured by a sandwich enzyme-linked immunosorbent assay (ELISA), and the concentration of the analytes were back-calculated with Watson LIMSTM V.7.3.0.01 (Thermo Scientific Inc.). The comparison of similarity between B and T were investigated with a balanced experiment design, and the results of the standard curves were evaluated with a statistic method, square of sum, using the statistic tool, Allfit. Systematic validation of the method from accuracy, precision, specificity, sensitivity, selectivity, dilution linearity, and stability etc. aspects. RESULTS The results of the standard curves from the balanced designed experiments of the biosimilar and the reference were compared, and the statistical result was P=0.050 7, which showed no significant statistic difference (P>0.05). The systematic validation results showed that the intra-assay accuracy and precision of the method between the biosimilar and the reference were within the range of 5.6%-7.7% and -9.0%--0.8%, respectively; the inter-assay accuracy and precision of the method between the B and the T were within the range of 6.5%-7.2% and -7.3%--3.2%, respectively; the total error of the method was not more than 13.9%, which met the methodological acceptance criteria. The results of selectivity, specificity, dilution linearity, and stability were within the acceptance criteria. CONCLUSION The ELISA method, which was systematically validated with the balanced experimental design meets the acceptance criteria of biosimilar methodology requirement, and could be used to support the pharmacokinetic study of rituximab and its biosimilar.
王梦佳, 董立厚, 李弯弯, 吕晓龙, 王变珍, 葛志强. 运用ELISA对利妥昔单抗及其生物类似药在方法学上进行相似性验证[J]. 中国药学杂志, 2020, 55(19): 1622-1628.
WANG Meng-jia, DONG Li-hou, LI Wan-wan, LÜ Xiao-long, WANG Bian-zhen, GE Zhi-qiang. Method Validation of Methodology Similarity between Rituximab and Its Biosimilar by ELISA. Chinese Pharmaceutical Journal, 2020, 55(19): 1622-1628.
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