中国新药杂志

治疗性单克隆抗体类药物分析技术及其应用

PDF下载

张国林 ¹ , 景荣先 ² , 郭乐 ³ , 邢以文 ¹ , 薛满 ¹

中国新药杂志 | 综述 2021,30(6): 528-534

收起

中国新药杂志 |综述 2021 , 30 (6) : 528 -534

治疗性单克隆抗体类药物分析技术及其应用

全屏

张国林¹, 景荣先², 郭乐³, 邢以文¹, 薛满¹

作者信息

¹苏州市药品检验检测研究中心，苏州　215000

²苏州市立医院，苏州　215000

³宁夏医科大学，银川　750004

作者简介:

张国林，男，博士，副主任药师，主要从事药品检测质量控制方面的研究。联系电话：（0512）66090229，E-mail：zhangguolin2006@163.com。

Analysis technology and application progress of therapeutic monoclonal antibody drugs

Guo-lin ZHANG¹, Rong-xian JING², Le GUO³, Yi-wen XING¹, Man XUE¹

Affiliations

¹Suzhou Institute for Drug Control, Suzhou 215000, China

²Suzhou Municipal Hospital, Suzhou 215000, China

³Ningxia Medical University, Yinchuan 750004, China

出版时间: 2021-03-30

文章导航

摘要

收起

治疗性单克隆抗体（单抗）类药物是高度均一的生物大分子，抗原表位、理化性质和生物活性等方面的均一性程度高，能够通过特异性地与靶标分子结合发挥治疗作用。单抗类药物治疗过程中不良反应发生率低、患者耐受性高。单抗类药物是生物药物领域的热点和焦点，已成为现代生物制药行业中占比最大、增长最快的细分领域。根据生产技术和工艺的不同，单抗类药物发展已经经历了4代。我国最近批准上市的信迪利单抗注射液、特瑞普利单抗注射液、注射用卡瑞利珠单抗和替雷利珠单抗注射液等均属于第3代人源化单克隆抗体药物。单抗类药物分子结构复杂，容易发生二聚体、多聚体、末端氨基酸突变等不均一性变化，严重影响临床用药的安全性和有效性。对单抗类药物的原液和成品进行有效的质量控制是确保该类产品安全性和有效性的重要措施。目前，单抗类产品质量控制与分析的主要方法包括高效液相色谱（HPLC）法、酶联免疫吸附测定（ELISA）法、毛细管区带电泳（CZE）法、毛细管等电聚焦电泳（cIEF）法、成像毛细管等电聚焦电泳（iCIEF）法和十二烷基硫酸钠-毛细管电泳（CE-SDS）法等。本文就单抗类药物质量控制分析方法及应用进行综述，为相关研究及产品质量控制提供参考。

关键词

单克隆抗体 / 高效液相色谱法 / 酶联免疫吸附测定法 / 毛细管区带电泳法 / 细管等电聚焦电泳法 / 十二烷基硫酸钠-毛细管电泳

Abstract

收起

Therapeutic monoclonal antibody (mAb) drugs are highly homogeneous bio-macromolecules, with high degree of homogeneity in antigen epitopes, physical and chemical properties and biological activities, which can specifically bind to the target molecules. In the course of treatment, the incidence of adverse reactions is low while the tolerance of patients is high. Monoclonal antibodies are the focus in the field of bio-pharmaceutics, which have become the most important and fastest growing area in modern biopharmaceutical industry. According to the different production process and technology, the development of monoclonal antibodies has experienced four generations. Sintilimab injection, treprizumab injection, carrizumab for injection and tirizumab injection recently approved in China are all the third generation of humanized monoclonal antibody drugs. The molecular structure of monoclonal antibody drugs is complex, which is easy to change the heterogeneity of dimer, polymer and terminal amino acid mutation. These changes may cause serious effects on the safety and effectiveness of clinical medication. It is an important measure to control the quality of the original and finished products of monoclonal anti-tumor drugs to ensure the safety and effectiveness of such products. At present, the main methods of quality control and analysis of monoclonal antibody products include HPLC, ELISA, CZE, cIEF, iCIEF and CE-SDS. In this paper, the analysis methods for quality control and application status of monoclonal antibodies were reviewed, which can provide reference for related research and product quality control.

Key words

monoclonal antibody / high performance liquid chromatography / enzyme linked immunosorbent assay / capillary zone electrophoresis / capillary isoelectric polycarbonate electrophoresis / sodium dodecyl sulfate capillary electrophoresis

引用本文

张国林, 景荣先, 郭乐, 邢以文, 薛满. 治疗性单克隆抗体类药物分析技术及其应用. 中国新药杂志, 2021 , 30 (6) : 528 -534 .

Guo-lin ZHANG, Rong-xian JING, Le GUO, Yi-wen XING, Man XUE. Analysis technology and application progress of therapeutic monoclonal antibody drugs[J]. Chinese Journal of New Drugs, 2021 , 30 (6) : 528 -534 .

基金

收起

江苏省市场监督管理局科技项目(KJ207561)
苏州市科技发展计划资助项目(SYS201425)

参考文献

收起

文献

收起

参考文献引证文献

排序方式：

[1]

TAKAHASHI Y, YASUI T, MINAMI K, et al. Radiation enhances the efficacy of antitumor immunotherapy with an immunocomplex of interleukin-2 and its monoclonal antibody[J]. Anticancer Res, 2017, 37(12): 6799-6806.

[2]

NAPOLITANO S, MARTINI G, MARTINELLI E, et al. Antitumor efficacy of triple monoclonal antibody inhibition of epidermal growth factor receptor (EGFR) with MM151 in EGFR-dependent and in cetuximab-resistant human colorectal cancer cells[J]. Oncotarget, 2017, 8(47): 82773-82783.

[3]

JUTATHIS K, PONGKITWITOON B, SRITULARAK B, et al. Development of monoclonal antibody-based enzyme-linked immunosorbent assay for quantitative quality control of derris scandens (roxb.) benth[J]. J Immunoassay Immunochem, 2019, 40(4): 407-418.

[4]

NEBIJA D, NOE CR, URBAN E, et al. Quality control and stability studies with the monoclonal antibody, trastuzumab: Application of 1D- vs. 2D-gel electrophoresis[J]. Int J Mol Sci, 2014, 15(4): 6399-6411.

[5]

KATO Y, OHISHI T, YAMADA S, et al. Anti-human epidermal growth factor receptor 2 monoclonal antibody H₂Mab -41 exerts antitumor activity in a mouse xenograft model of colon cancer[J]. Monoclon Antib Immunodiagn Immunother, 2019, 38(4): 157-161.

[6]

辛中帅, 张彦彦, 晋小雁, 等. 我国已上市治疗用单抗类产品分析[J]. 中国药事, 2019, 33(9): 986-990.

[7]

YIN Z, GAO M, CHU S, et al. Antitumor activity of a newly developed monoclonal antibody against ROR1 in ovarian cancer cells[J]. Oncotarget, 2017, 8(55): 94210-94222.

[8]

THEY L, MICHAUD HA, BECQUART O, et al. PD-1 blockade at the time of tumor escape potentiates the immune-mediated antitumor effects of a melanoma-targeting monoclonal antibody[J]. Oncoimmunology, 2017, 6(10): e1353857.

[9]

朱丹, 李月阳, 宋燕青, 等. PD-1抑制剂信迪利单抗的临床研究进展[J]. 中国医院药学杂志, 2020, 40(1): 120-123.

[10]

唐碧霞, 斯璐, 郭军. 特瑞普利单抗治疗黑色素瘤的适应证分析[J]. 中国肿瘤临床, 2019, 46(16): 857-860.

[11]

FADERL S, FERRAJOLI A, WIERDA W, et al. Alemtuzumab by continuous intravenous infusion followed by subcutaneous injection plus rituximab in the treatment of patients with chronic lymphocytic leukemia recurrence[J]. Cancer, 2010, 116(10): 2360-2365.

[12]

DING S, XIONG J, LEI D, et al. Recombinant nanocomposites by the clinical drugs of Abraxane^® and Herceptin^® as sequentially dual-targeting therapeutics for breast cancer[J]. J Cancer, 2018, 9(3): 502-511.

[13]

CHEN P, LI X, ZHANG R, et al. Combinative treatment of β-elemene and cetuximab is sensitive to KRAS mutant colorectal cancer cells by inducing ferroptosis and inhibiting epithelial-mesenchymal transformation[J]. Theranostics, 2020, 10(11): 5107-5119.

[14]

PATIL VM, NORONHA V, JOSHI A, et al. A randomized phase 3 trial comparing nimotuzumab plus cisplatin chemoradiotherapy versus cisplatin chemoradiotherapy alone in locally advanced head and neck cancer[J]. Cancer, 2019, 125(18): 3184-3197.

[15]

YOSHIDA H, KIM YH, SAKAMORI Y, et al. A randomized phase II study of maintenance bevacizumab, pemetrexed or bevacizumab plus pemetrexed for advanced non-squamous non-small cell lung cancer[J]. Anticancer Res, 2020, 40(5): 2981-2987.

[16]

LIU J, WANG CY, CAO L, et al. Treatment of advanced intrahepatic cholangiocarcinoma with sintilimab combined with tegafur-gimeracil-oteracil potassium capsules (S-1): a case report[J]. Ann Palliat Med, 2020, 9(2): 497-503.

[17]

国家药品监督管理局. 治疗霍奇金淋巴瘤的首个国产PD-1抗体药物信迪利单抗注射液获批上市[EB/OL]. (2018-12-28). http://www.nmpa.gov.cn/WS04/CL2056/334158.html.

[18]

国家药品监督管理局. 首个国产PD-1抗体药物特瑞普利单抗注射液获批上市[EB/OL]. (2018-12-17). http://www.nmpa.gov.cn/WS04/CL2094/333924.html.

[19]

国家药品监督管理局. 2020年03月05日药品批件发布通知[EB/OL]. (2018-3-5). http://www.nmpa.gov.cn/WS04/CL2455/375293.html.

[20]

国家药品监督管理局. 2020年04月13日药品批件发布通知[EB/OL]. (2020-4-13). http://www.nmpa.gov.cn/WS04/CL2455/376455.html.

[21]

WANG WH, CHEUNG-LAU J, CHEN Y, et al. Specific and high-resolution identification of monoclonal antibody fragments detected by capillary electrophoresis-sodium dodecyl sulfate using reversed-phase HPLC with top-down mass spectrometry analysis[J]. mAbs, 2019, 11(7): 1233-1244.

[22]

NAVAS N, HERRERA A, MARTÍNEZ-ORTEGA A, et al. Quantification of an intact monoclonal antibody, rituximab, by (RP) HPLC/DAD in compliance with ICH guidelines[J]. Anal Bioanal Chem, 2013, 405(29): 9351-9363.

[23]

GUAN YG, WU XL, MENG HC. Indirect competitive ELISA based on monoclonal antibody for the detection of 5-hydroxymethyl-2-furfural in milk, compared with HPLC[J]. J Dairy Sci, 2013, 96(8): 4885-4890.

[24]

BOND MD, PANEK ME, ZHANG ZR, et al. Evaluation of a dual-wavelength size exclusion HPLC method with improved sensitivity to detect protein aggregates and its use to better characterize degradation pathways of an IgG1 monoclonal antibody[J]. J Pharm Sci, 2010, 99(6): 2582-2597.

[25]

秦海艳, 安晨, 宋兰兰, 等. 阳离子交换高效液相色谱检测重组人源化抗CD52单克隆抗体的电荷异质性[J]. 微生物学免疫学进展, 2018, 46(4): 19-25.

[26]

柳军凯, 贺鹏飞, 赖燕华, 等. 人乳头瘤病毒18型L1抗原的肽图特征峰分析[J]. 药物分析杂志, 2019, 39(10): 1836-1843.

[27]

郭莎, 王开芹, 武刚, 等. 抗PD-L1单抗的质量控制研究[J]. 药物分析杂志, 2019, 39(1): 13-22.

[28]

付志浩, 陈伟, 李萌, 等. 重组抗CD38单克隆抗体的质量研究[J]. 药物分析杂志, 2019, 39(1): 23-29.

[29]

乔玉玲, 黄铮, 秦海艳, 等. 抗CD52单克隆抗体HPLC-肽图分析方法的建立[J]. 生物技术通报, 2018, 34(11): 216-222.

[30]

郭秋菊, 李雪, 徐志远, 等. 重组人源化抗程序性死亡受体-1单克隆抗体关键质控方法的建立[J]. 中国生物制品学杂志, 2018, 31(4): 376-382.

[31]

ORBEGOZO-MEDINA RA, MARTÍNEZ-SERNÁNDEZ V, PERTEGUER MJ, et al. In-plate recapturing of a dual-tagged recombinant Fasciola antigen (FhLAP) by a monoclonal antibody (US9) prevents non-specific binding in ELISA[J]. PLoS One, 2019, 14(2): e0211035.

[32]

PRASATKAEW W, NANTHANAWAT P, KHONGCHAREONPORN N, et al. A monoclonal antibody against lates calcarifer vitellogenin and a competitive ELISA to evaluate vitellogenin induction after exposure to xenoestrogen[J]. J Environ Sci (China), 2019, 75(11): 325-333.

[33]

詹明硕, 刘方杰, 张亚兰. 噬菌体抗体库技术筛选抗VEGF165单克隆抗体[J]. 生物技术通讯, 2019, 30(3): 348-353.

[34]

宋媛媛. 抗肿瘤药物奥希替尼和重组人鼠嵌合型抗CD20单克隆抗体的药物浓度分析和药效学研究[D]. 北京协和医学院, 2018.

[35]

乔玉玲, 黄峥, 秦海艳, 等. 抗CD52单克隆抗体中残留蛋白AELISA定量检测方法的验证[J]. 微生物学免疫学进展, 2018, 46(1): 25-29.

[36]

郑逢佳, 贾志君, 谢新遥, 等. 抗体类药物的生物分析方法研究概况[J]. 中国临床药理学与治疗学, 2020, 25(2): 227-232.

[37]

RUSTANDI RR, WANG Y. Use of CE-SDS gel for characterization of monoclonal antibody hinge region clipping due to copper and high pH stress[J]. Electrophoresis, 2011, 32(21): 3078-3084.

[38]

王建锋, 乔玉玲, 秦海燕, 等. 应用CE-SDS测定抗CD52人源化单克隆抗体参比品单体纯度及非糖化重链比例[J]. 现代生物医学进展, 2018, 18(15): 2836-2840, 2828.

[39]

陈泓序, 屈锋. 毛细管电泳技术在单克隆抗体药物分析中的应用[J]. 色谱, 2018, 36(3): 195-208.

[40]

SÄNGER-VAN DE GRIEND CE. CE-SDS method development, validation, and best practice-An overview[J]. Electrophoresis, 2019, 40(18-19): 2361-2374.

[41]

KAHLE J, MAUL KJ, WÄTZIG H. The next generation of capillary electrophoresis instruments: Performance of CE-SDS protein analysis[J]. Electrophoresis, 2018, 39(2): 311-325.

[42]

ESTERMAN AL, KATIYAR A, KRISHNAMURTHY G. Implementation of USP antibody standard for system suitability in capillary electrophoresis sodium dodecyl sulfate (CE-SDS) for release and stability methods[J]. J Pharm Biomed Anal, 2016, 128: 447-454.

[43]

DADA OO, RAO R, JONES N, et al. Comparison of SEC and CE-SDS methods for monitoring hinge fragmentation in IgG1 monoclonal antibodies[J]. J Pharm Biomed Anal, 2017, 145: 91-97.

[44]

DAI J, LAMP J, XIA QW, et al. Capillary isoelectric focusing-mass spectrometry method for the separation and online characterization of intact monoclonal antibody charge variants[J]. Anal Chem, 2018, 90(3): 2246-2254.

[45]

HOSKEN BD, LI C, MULLAPPALLY B, et al. Isolation and characterization of monoclonal antibody charge variants by free flow isoelectric focusing[J]. Anal Chem, 2016, 88(11): 5662-5669.

[46]

SUBA D, URBÁNYI Z, SALGÓ A. Capillary isoelectric focusing method development and validation for investigation of recombinant therapeutic monoclonal antibody[J]. J Pharm Biomed Anal, 2015, 114: 53-61.

[47]

NICHOLS P, LI L, KUMAR S, et al. Rational design of viscosity reducing mutants of a monoclonal antibody: hydrophobic versus electrostatic inter-molecular interactions[J]. mAbs, 2015, 7(1): 212-230.

[48]

王文波, 武刚, 于传飞, 等. 治疗性单克隆抗体电荷异质性分析方法比较[J]. 药物分析杂志, 2017, 37(8): 1383-1388.

[49]

刘振东, 韩国华, 杨勇, 等. 全柱成像毛细管等电聚焦(iCIEF)技术分析重组人IgG VEGFR: Fc-融合蛋白电荷异质性[J]. 药物分析杂志, 2018, 38(3): 539-544.

[50]

孟晓光, 甄里, 刘悦玫, 等. 全柱成像毛细管等电聚焦电泳分析蛋白质药物电荷异质性[J]. 分析测试学报, 2017, 36(3): 343-348.

[51]

LIN J, LAZAR AC. Determination of charge heterogeneity and level of unconjugated antibody by imaged cIEF[J]. Methods Mol Biol, 2013, 1045: 295-302.

[52]

ZHU GJ, SUN LL, WOJCIK R, et al. A rapid cIEF-ESI-MS/MS method for host cell protein analysis of a recombinant human monoclonal antibody[J]. Talanta, 2012, 98: 253-256.

[53]

王文波, 于传飞, 张峰, 等. 基于毛细管区带电泳的IgG2型单抗电荷异质性分析方法的建立[J]. 中国药学杂志, 2018, 53(8): 627-631.

[54]

陈继军, 南建军, 乔玉玲, 等. 重组抗CD52单克隆抗体亲和层析纯化工艺的优化[J]. 中国生物制品学杂志, 2019, 32(4): 439-444.

[55]

赵祖波, 陈继军, 赵晓瑞, 等. 抗CD52单克隆抗体外源DNA残留量检测方法的建立及验证[J]. 中国生物制品学杂志, 2017, 30(12): 1305-1310.

2021年第30卷第6期

PDF下载

引用本文

BibTeX

文章信息

首发时间：2026-10-29
出版时间：2021-03-30

补充材料

相关文章

文章信息

作者

出版历史

录用日期：2020-04-07

基金

江苏省市场监督管理局科技项目(KJ207561)

苏州市科技发展计划资助项目(SYS201425)

作者信息

¹苏州市药品检验检测研究中心，苏州　215000

²苏州市立医院，苏州　215000

³宁夏医科大学，银川　750004

参考文献

分享链接

https://castjournals.cast.org.cn/joweb/zgxyzz/CN/1322627634878599628

分享至

全文二维码

扫描看全文

引用本文

BibTeX

本文的引用情况

2种不同金属材料的力学参数

科 Family	属数 Number of genus	种数 Number of species	占总种数比例 Percentage of total species (%)	属 Genus	种数 Number of species	占总种数比例 Percentage of total species (%)
鹅膏菌科Amanitaceae	2	11	5.26	鹅膏菌属 Amanita	10	4.78
小菇科 Mycenaceae	2	12	5.74	丝盖伞属 Inocybe	5	2.39
多孔菌科 Polyporaceae	8	14	6.70	蜡蘑属 Laccaria	5	2.39
红菇科 Russulaceae	3	23	11.00	小皮伞属 Marasmius	6	2.87
				小菇属 Mycena	11	5.26
				光柄菇属 Pluteus	5	2.39
				红菇属 Russula	17	8.13
				栓菌属 Trametes	5	2.39

关闭全屏

BibTeX
EndNote
RefWorks
TxT