Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates

doi:10.1038/nprot.2017.058

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	Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates
	Tang, Feng 2,3; Wang, Lai-Xi 1; Huang, Wei 2,3
刊名	NATURE PROTOCOLS
	2017-08
卷号	12 期号:8 页码:1702-1721
ISSN号	1754-2189
DOI	10.1038/nprot.2017.058
文献子类	Article
英文摘要	Glycoengineered therapeutic antibodies and glycosite-specific antibody-drug conjugates (gsADCs) have generated great interest among researchers because of their therapeutic potential. Endoglycosidase-catalyzed in vitro glycoengineering technology is a powerful tool for IgG Fc (fragment cystallizable) N-glycosylation remodeling. In this protocol, native heterogeneously glycosylated IgG N-glycans are first deglycosylated with a wild-type endoglycosidase. Next, a homogeneous N-glycan substrate, presynthesized as described here, is attached to the remaining N-acetylglucosamine (GlcNAc) of IgG, using a mutant endoglycosidase (also called endoglycosynthase) that lacks hydrolytic activity but possesses transglycosylation activity for glycoengineering. Compared with in vivo glycoengineering technologies and the glycosyltransferase-enabled in vitro engineering method, the current approach is robust and features quantitative yield, homogeneous glycoforms of produced antibodies and ADCs, compatibility with diverse natural and non-natural glycan structures, convenient exploitation of native IgG as the starting material, and a well-defined conjugation site for antibody modifications. Potential applications of this method cover a broad scope of antibody-related research, including the development of novel glycoengineered therapeutic antibodies with enhanced efficacy, site-specific antibody-drug conjugation, and site-specific modification of antibodies for fluorescent labeling, PEGylation, protein cross-linking, immunoliposome formation, and so on, without loss of antigen-binding affinity. It takes 5-8 d to prepare the natural or modified N-glycan substrates, 3-4 d to engineer the IgG N-glycosylation, and 2-5 d to synthesize the small-molecule toxins and prepare the gsADCs.
资助项目	National Natural Science Foundation of China (NNSFC)[21372238] ; National Natural Science Foundation of China (NNSFC)[21572244] ; SIMM institute fund[CASIMM0120153004] ; 'Personalized Medicines: Molecular Signature-based Drug Discovery and Development' Strategic Priority Research Program of the Chinese Academy of Sciences[XDA12020311]
WOS关键词	DEPENDENT CELLULAR CYTOTOXICITY ; IN-VITRO GALACTOSYLATION ; HIGH-MANNOSE GLYCANS ; FC-GAMMA-RIII ; ANTIINFLAMMATORY ACTIVITY ; THERAPEUTIC ANTIBODIES ; N-ACETYLGLUCOSAMINE ; EFFECTOR FUNCTIONS ; IMMUNOGLOBULIN-G ; COPPER-FREE
WOS研究方向	Biochemistry & Molecular Biology
语种	英语
出版者	NATURE PUBLISHING GROUP
WOS记录号	WOS:000406408200003
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/272548]
专题	生物技术药物研发中心（筹）药物安全性评价中心
通讯作者	Huang, Wei
作者单位	1.Univ Maryland, Dept Chem & Biochem, College Pk, MD 20742 USA 2.Univ Chinese Acad Sci, Beijing, Peoples R China; 3.Chinese Acad Sci, Shanghai Inst Mat Med, CAS Ctr Excellence Mol Cell Sci, CAS Key Lab Receptor Res, Shanghai, Peoples R China;
推荐引用方式 GB/T 7714	Tang, Feng,Wang, Lai-Xi,Huang, Wei. Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates[J]. NATURE PROTOCOLS,2017,12(8):1702-1721.
APA	Tang, Feng,Wang, Lai-Xi,&Huang, Wei.(2017).Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates.NATURE PROTOCOLS,12(8),1702-1721.
MLA	Tang, Feng,et al."Chemoenzymatic synthesis of glycoengineered IgG antibodies and glycosite-specific antibody-drug conjugates".NATURE PROTOCOLS 12.8(2017):1702-1721.