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Specific Conjugation of the Hinge Region for Homogeneous Preparation of Antibody Fragment-Drug Conjugate: A Case Study for Doxorubicin-PEG-anti-CD20 Fab' Synthesis
Zhou, Zhan1,2; Zhang, Jing1; Zhang, Yan1; Ma, Guanghui1; Su, Zhiguo1
2016
Source PublicationBIOCONJUGATE CHEMISTRY
ISSN1043-1802
Volume27Issue:1Pages:238-246
AbstractConventional preparation strategies for antibody-drug conjugates (ADCs) result in heterogeneous products with various molecular sizes and species. In this study, we developed a homogeneous preparation strategy by site-specific conjugation of the anticancer drug with an antibody fragment. The model drug doxorubicin (DOX) was coupled to the Fab' fragment of anti-CD20 IgG at its permissive sites through a heterotelechelic PEG linker, generating an antibody fragment-drug conjugate (AFDC). Anti-CD20 IgG was digested and reduced specifically with beta-mercaptoethylamine to generate the Fab' fragment with two free mercapto groups in its hinge region. Meanwhile, DOX was conjugated with alpha-succinimidylsuccinate omega-maleimide polyethylene glycol (NHS-PEG-MAL) to form MAL-PEG-DOX, which was subsequently linked to the free mercapto containing Fab' fragment to form a Fab'-PEG-DOX conjugate. The dual site-specific bioconjugation was achieved through the combination of highly selective reduction of IgG and introduction of heterotelechelic PEG linker. The resulting AFDC provides an utterly homogeneous product, with a definite ratio of one fragment to two drugs. Laser confocal microscopy and cell ELISA revealed that the AFDC could accumulate in the antigen-positive Daudi tumor cell. In addition, the Fab'-PEG-DOX retained appreciable targeting ability and improved antitumor activity, demonstrating an excellent therapeutic effect on the lymphoma mice model for better cure rate and significantly reduced side effects.
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
DOI10.1021/acs.bioconjchem.5b00626
Indexed BySCI
Language英语
WOS KeywordMONOCLONAL-ANTIBODY ; THERAPEUTIC INDEX ; CANCER-THERAPY ; DOXORUBICIN ; EXPRESSION ; ANTI-CD20 ; GROWTH ; POTENT ; CELLS
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Chemistry, Multidisciplinary ; Chemistry, Organic
Funding OrganizationNational Natural Science Foundation of China(21106163 ; Science and Technology Service Network Initiative(KFJ-EW-STS-027) ; National High Technology Research and Development Program of China (863 Program)(2012AA021202 ; National Key Basic Research Program of China (973 Program)(2013CB733600) ; National Key Foundation for Exploring Scientific Instrument of China(2013YQ14040508) ; 21306207 ; 2012AA02A406 ; 21336010) ; 2014AA021006)
WOS IDWOS:000368651600026
Citation statistics
Cited Times:4[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.ipe.ac.cn/handle/122111/20598
Collection生化工程国家重点实验室
Affiliation1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, 1 Beierjie St, Beijing 100190, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Zhou, Zhan,Zhang, Jing,Zhang, Yan,et al. Specific Conjugation of the Hinge Region for Homogeneous Preparation of Antibody Fragment-Drug Conjugate: A Case Study for Doxorubicin-PEG-anti-CD20 Fab' Synthesis[J]. BIOCONJUGATE CHEMISTRY,2016,27(1):238-246.
APA Zhou, Zhan,Zhang, Jing,Zhang, Yan,Ma, Guanghui,&Su, Zhiguo.(2016).Specific Conjugation of the Hinge Region for Homogeneous Preparation of Antibody Fragment-Drug Conjugate: A Case Study for Doxorubicin-PEG-anti-CD20 Fab' Synthesis.BIOCONJUGATE CHEMISTRY,27(1),238-246.
MLA Zhou, Zhan,et al."Specific Conjugation of the Hinge Region for Homogeneous Preparation of Antibody Fragment-Drug Conjugate: A Case Study for Doxorubicin-PEG-anti-CD20 Fab' Synthesis".BIOCONJUGATE CHEMISTRY 27.1(2016):238-246.
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