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期刊名称:药物分析杂志
主管单位:中国科学技术协会
主办单位:中国药学会
承办:中国食品药品检定研究院
主编:金少鸿
地址:北京天坛西里2号
邮政编码:100050
电话:010-67012819,67058427
电子邮箱:ywfx@nicpbp.org.cn
国际标准刊号:ISSN 0254-1793
国内统一刊号:CN 11-2224/R
邮发代号:2-237
 

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重组抗CD38单克隆抗体的质量研究

Quality study of recombinant anti-CD38 monoclonal antibody

作者(英文):
分类号:R917
出版年·卷·期(页码):2019,39 (1):23-29
DOI: 10.16155/j.0254-1793.2017.01.01
-----摘要:-------------------------------------------------------------------------------------------

目的:针对抗白细胞分化抗原38(cluster of differentiation 38,CD38)单抗的关键质量属性建立质控方法。方法:采用肽图法进行鉴别;采用还原/非还原十二烷基磺酸钠毛细管电泳(capillary electrophoresis-sodium dodecyl sulfonate,CE-SDS)和尺寸排阻色谱(size exclusion-high performance liquid chromatography,SEC-HPLC)进行纯度控制;采用成像毛细管等电聚焦电泳(imaging capillary isoelectric focusing electrophoresis,iCIEF)测定电荷异质性;采用酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)测定结合活性;采用补体依赖的细胞毒作用(complement dependent cytotoxicity,CDC)和抗体依赖的细胞介导的细胞毒作用(antibody-dependent cell-mediated cytotoxicity,ADCC)的方法测定生物学活性。其他各项指标均应符合2015年版《中华人民共和国药典》以及其他相关要求。结果:肽图检测具有特征图谱,起到很好的鉴别作用。还原CE-SDS的轻链与重链峰面积之和百分比为(97.87±0.72)%,非还原CE-SDS主峰峰面积百分比为(97.70±0.35)%;SEC-HPLC单体的峰面积百分比为(99.15±0.01)%,高分子量物质的峰面积百分比为(0.47±0.002)%,低分子量物质的峰面积百分比为(0.37±0.01)%。iCIEF分析的主要亚型的峰面积百分比为(63.54±0.37)%,酸性亚型的峰面积百分比为(23.07±0.50)%,碱性亚型的峰面积百分比为(13.38±0.12)%。结合活性的半最大效应浓度(concentration for 50% of maximal effect,EC50)值为(7.35±0.06)ng·mL-1。CDC活性EC50值为(270.67±12.42)ng·mL-1,ADCC活性EC50值为(8.71±1.04)ng·mL-1结论:针对抗CD38单抗的质量属性,研究建立质控方法并进行质量研究,确保产品的安全、有效和质量可控,对抗CD38单抗的质量控制具有借鉴意义。

-----英文摘要:---------------------------------------------------------------------------------------

Objective:To establish the quality control methods for the key quality attributes of the anti-leukocyte differentiation antigen 38 (cluster of differentiation 38,CD38) monoclonal antibody(mAb). Methods:Peptide map analysis was used for identification of an anti-CD38 mAb. The purity was measured by reduced/non-reduced capillary electrophoresis-sodium dodecyl sulfonate(CE-SDS) and size exclusion-high performance liquid chromatography(SEC-HPLC). Charge heterogeneity was measured by imaging capillary isoelectric focusing electrophoresis (iCIEF). Binding potency was measured by ELISA assay. The biological activity was determined by complement dependent cytotoxicity (CDC) method and antibody-dependent cell-mediated cytotoxicity (ADCC) method. Other quality control items complied with corresponding requirements of Chinese Pharmacopeia 2015 edition and related guidelines. Results:Peptide mapping showed the specific spectrum of the anti-CD38 mAb,which could be adopted for the identification test. The sum percentage of light chain and heavy chain peak area detected by reduced CE-SDS was (97.87±0.72)%;the area percentage of main peak detected by non-reduced CE-SDS was (97.70±0.35)%. The area percentage of monomer peak,high molecular mass peak and low molecular mass peak detected by SEC-HPLC were (99.15±0.01)%,(0.47±0.002)% and (0.37±0.01)%,respectively. The area percentage of major subtypes peak,acidic subtypes peak and alkaline subtypes peak detected by iCIEF were (63.54±0.37)%,(23.07±0.50)% and (13.38±0.12)%,respectively. The concentration for 50% of maximal effect (EC50) of binding activity was (7.35±0.06)ng·mL-1. The EC50 value of CDC activity was (270.67±12.42)ng·mL-1 and the EC50 value of ADCC activity was (8.71±1.04)ng·mL-1. Conclusion:A series of quality control methods for anti-CD38 mAb have been established,which can be adaptable in ensuring the safety,effectiveness and quality controllability for the product. It has reference significance for the quality control of anti-CD38 mAb. 

-----参考文献:---------------------------------------------------------------------------------------

[1] REINHERZ EL,KUNG PC,GOLDSTEIN G,et al.Discrete stages of human intrathymic differentiation:analysis of normal thymocytes and leukemic lymphoblasts of T cell lineage[J].Proc Natl Acad Sci USA,1980,77(3):1588
[2] HOWARD M,GRIMALDI JC,BAZAN JF,et al.Formation and hydrolysis of cyclic ADP-ribose catalyzed by lymphocyte antigen CD38[J].Science,1993,262(5136):1056
[3] CAMPANA D,SUZUKI T,TODISCO E,et al.CD38 in hematopoiesis[J].Chem Immunol,2000,75(9):169
[4] LIN P,OWENS R,TRICOT G,et al.Flow cytometricimmunophenotypic analysis of 306 cases of multiple myeloma[J].Am J Clin Pathol,2004,121(4):482
[5] SCHWONZEN M,POHL C,STEINMETZ T,et al.Immunophenotyping of low-grade B-cell lymphoma in blood and bone marrow:poor correlation between immune phenotype and cytological/histological classification[J].Br J Haematol,1993,83(2):232
[6] DOMINGO-DOMèNECH E,DOMINGO-CLARóS A,GONZáLEZ-BARCA E,et al.CD38 expression in B-chronic lymphocytic leukemia:association with clinical presentation and outcome in 155 patients[J].Haematologica,2002,87(10):1021
[7] KUMAR SK,LEE JH,LAHUERTA JJ,et al.Risk of progression and survival in multiple myeloma relapsing after therapy with IMiDs and bortezomib:a multicenter international myeloma working group study[J].Leukemia,2012,26(1):149
[8] USMANI S,AHMADI T,NG Y,et al.Analysis of real-world data on overall survival in multiple myeloma patients with ≥ 3 prior lines of therapy including a proteasome inhibitor (PI) and an immune modulatory drug (IMiD),or double refractory to a PI and an IMiD[J].Oncologist,2016,21:1335
[9] JIAN H,JIE J,YAN X,et al.Randomized,double-blind,placebo-controlled phase 3 study of ixazomib plus lenalidomide-dexamethasone in patients with relapsed/refractory multiple myeloma:China continuation study[J].J Hematol Oncol,2017,10:137
[10] BLAIR HA.Daratumumab:a review in relapsed and/or refractory multiple myeloma[J].Drugs,2017,77(18):2013
[11] TAMURA H.Immunopathogenesis and appropriate use of monoclonal antibody agents in multiple myeloma[J].Rinsho Ketsueki,2018,59(10):2169
[12] van de DONK NWCJ,USMANI SZ.CD38 antibodies in multiple myeloma:mechanisms of action and modes of resistance[J].Front Immunol,2018,9:2134
[13] 王军志.生物技术药物研究开发和质量控制[M].第3版.北京:科学出版社,2018 WANG JZ.Research,Development and Quality Control of Biopharmaceuticals[M].3rd Ed.Beijing:Science Press,2018
[14] ESPINOSA-de la GARZA CE,PERDOMO-ABúNDEZ FC,PADILLA-CALDERóN J,et al.Analysis of recombinant monoclonal antibodies by capillary zone electrophoresis[J].Electrophoresis,2013,34(8):1133
[15] CAO J,SUN W,GONG F,et al.Charge profiling and stability testing of biosimilar by capillary isoelectric focusing[J].Electrophoresis,2014,35(10):1461
[16] WU G,YU C,WANG W,et al.Interlaboratory method validation of icIEF methodology for analysis of monoclonal antibodies[J].Electrophoresis,2018,39(16):2091
[17] DU Y,WALSH A,EHRICK R,et al.Chromatographic analysis of the acidic and basic species of recombinant monoclonal antibodies[J].MAbs,2012,4(5):578
[18] CHENG ZJ,GARVIN D,PAGUIO A.Development of a robust reporter-based ADCC assay with frozen,thaw-and-use cells to measure Fc effector function of therapeutic antibodies[J].J Immunol Methods,2014,414:69
[19] LEI C,GONG R,YING T.Editorial:antibody Fc engineering:towards better therapeutics[J].Front Immunol,2018,9:2450
[20] READ EK,PARK JT,BRORSON KA.Industry and regulatory experience of the glycosylation of monoclonal antibodies[J].Biotechnol Appl Biochem,2011,58(4):213
[21] JEFFERIS R.Glycosylation as a strategy to improve antibody-based therapeutics[J].Nat Rev Drug Discov,2009,8(3):226
[22] YU C,GAO K,ZHU L,et al.At least two Fc Neu5Gc residues of monoclonal antibodies are required for binding to anti-Neu5Gc antibody[J].Sci Rep,2016,7:20029 

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