目的 验证受试品(抗人CD20嵌合单克隆抗体,biosimilar,B)与参比品(利妥昔单抗,rituximab,美罗华,reference,R)在方法学上的类似性。方法 受试品和参比品的测定采用双抗夹心酶联免疫吸附分析法(ELISA),检测结果用Watson LIMSTM V.7.3.0.01(Thermo Scientific公司)软件进行处理,方法类似性比较采用均衡实验设计并用Allfit软件中平方和的统计方式比较标准曲线类似性,方法从准确度、精密度、特异性、灵敏度、选择性、稀释线性及稳定性等方面进行系统验证。结果 受试品与参比品标准曲线平衡实验结果进行类似性比较,统计结果P=0.050 7无统计学差异(P>0.05);系统性验证结果表明,受试品和参比品批内准确度和精密度分别在5.6%～7.7%,-9.0%～-0.8%内,受试品和参比品批间准确度和精密度分别在6.5%～7.2%,-7.3%～-3.2%内,受试品和参比品的总误差≤13.9%,满足方法学接受标准;选择性、特异性、稀释线性和稳定性在方法学接受范围内。结论 经平衡实验设计和系统验证的ELISA法,满足生物类似物方法学接受标准,可用于利妥昔单抗生物类似药药动学研究。
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.
关键词
酶联免疫吸附分析法 /
单克隆抗体 /
生物类似药 /
利妥昔单抗 /
方法类似性验证
{{custom_keyword}} /
Key words
enzyme-linked immunosorbent assay /
monoclonal antibody /
biosimilar /
rituximab /
methodology similarity validation
{{custom_keyword}} /
中图分类号：
R917
{{custom_clc.code}}
({{custom_clc.text}})
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] CFDA. Guideline on Development and Evaluation of Biosimilars ( 2015) . 2015.
[2] EMA. Guideline on similar biological medicinal products. . (2005-10-30). http://www.ema.europa.eu/docs/en_GB/document_library/Scientific_guideline/2009/-09/WC500003517.pdf.
[3] GAO C Y, XIE S M, BAI Y, et al. VConsiderations on clinical study of biosimilars. Chin Pharm J(中国药学杂志), 2015,50(6):494-496.
[4] LIU B N, BAI Y, LUO J H. Biosimilarity study regarding product quality of candidate monoclonal antibodies as biosimilars. Chin Pharm J(中国药学杂志), 2017, 52(13):1194-1200.
[5] WHO. Guidelines on Evaluation of Similar Biotherapeutic Products (SBPs). . (2009-10). http://www.who int/biologicals/areas/biological_therepeutics/BIOTHERA-PEUTICS_TOR_WEB_22APRIL2010.pdf.
[6] FDA. Scientific considerations in demonstrating biosimilarity to a reference product. . (2015-04) . https://www.fda.gov/downloads/ drugs/guidance compliance regulat-ory information/guidances/ucm 291128.pdf.
[7] EMA. Guideline on similar biological medicinal products containing biotechnology-derived proteins as active substance: quality issues (revision 1) (EMA/CHMP/BWP/247713/2012) . (2015-04). . http://www. ema. europa. eu/docs/en_GB/document_librar-y/Scientific_guideline/2014/06/WC500167838. pdf.
[8] US Food and Drugs Administration. Scientific considerations in demonstrating biosimilarity to a reference product: Guidance for industry . 2015. https://www.fda.g-ov/downloads/ drugs/guidances/ucm291128.pdf. Accessed 31May2018.
[9] ANTONIA M S MÜLLER, IHORST G, MERTELSMANN R, et al. Epidemiology of non-Hodgkin′s lymphoma (NHL):trends, geographic distribution, and etiology. Ann Hematol, 2005, 84(1):1-12.
[10] HALLEK, MICHAEL. Chronic lymphocytic leukemia: 2013 update on diagnosis, risk stratification and treatment. Am J Hematol, 2013, 88(9):803-816.
[11] BANCHEREAU J. Human B lymphocytes: phenotype, proliferation, and differentiation. Adv Immunol, 1992, 52:125-262.
[12] MALONEY D G. Anti-CD20 antibody therapy for B-cell lymphomas. N Engl J Med, 2012, 366:2008-2016.
[13] ZELENETZ A D, GORDON L I, ABRAMSON, et al. NCCN guidelines insights: B-cell lymphomas, version 3.2019. J Natl Comprehensive Cancer Network, 2019, 17(6):650-661.
[14] DREYLING M, GHIELMINI M, RULE S, et al. Newly diagnosed and relapsed follicular lymphoma: ESMO clinical practice guidelines for diagnosis, treatment and follow-up. Ann Oncol, 2016, 27(15):83-90.
[15] International Pharmaceutical Regulators Forum Biosimilars working Group. Reflection paper on extrapolation of indications in authorization of biosimilar products version1. 3 . (2016). . www.i-p-r-f.org/files/1614/7315/4963/160725 IPRF BWG Reflection paper final draftver 1.3 IPRF MC Circulation.pdf.
[16] MARINI J C, ANDERSON M, CAI X Y, et al. Systematic verification of bioanalytical similarity between a biosimilar and a reference biotherapeutic: committee recommendations for the development and validation of a single ligand-binding assay to support pharmacokinetic assessments. AAPS, 2014, 16(6):1149-1158.
[17] FDA. Assay development and validation for immunogenicity testing of therapeutic protein products. . April 2016. https://www.fda.gov.
[18] UNITED STATES PHARMACOPEIA. Immunogenicity assays-design and validation of immunoassays to detect antidrug neutralizing antibody. . http://www.usp.org.
[19] SHANKAR G, DEVANARAYAN V, AMARAVADI L, et al. Recommendations for the validation of immunoassays used for detection of host antibodies against biotechnology products. J Pharm Biomed Anal, 2008, 48(5):1267-1281.
[20] GUPTA S, DEVANARAYAN V, FINCO D, et al. Recommendations for the validation of cell-based assays used for the detection of neutralizing antibody immune responses elicited against biological therapeutics. J Pharm Biomed Anal, 2011, 55(5):878-888.
[21] EMEA/CHMP/BMWP/14327/2006 Rev.1Guideline on immunogenicity assessment of biotechnology-derived therapeutic Proteins. . . https://www.ema.europa.eu/en/documents/scientific-guideline
[22] DESILVA B, SMITH W, WEINER R, et al. Recommendations for the bioanalytical method validation of ligand-binding assays to support pharmacokinetic assessments of macromolecules. Pharm Res, 2003, 20(11):1885-1900.
[23] CAI X Y, GOUTY D, BAUGHMAN S, et al. Recommendations and requirements for the design of bioanalytical testing used in comparability studies for biosimilar drug development. Bioanalysis, 2011, 3(5):535-540.
{{custom_fnGroup.title_cn}}
脚注
{{custom_fn.content}}
基金
国家“重大新药创制”科技重大专项资助 (2015ZX09501008)
{{custom_fund}}
PDF(4725 KB)
