Thermostability detection and optimization of glycoengineered antibodies and antibody-drug conjugates based on differential scanning flouremitry analysis

doi:10.1016/j.bioorg.2019.103391

	Thermostability detection and optimization of glycoengineered antibodies and antibody-drug conjugates based on differential scanning flouremitry analysis
	Qin, Ken 1,2; Shi, Wei 1,2; Zhao, Lei 1; Li, Mingjie 1; Tang, Yubo 1; Faridoon 1; Jiang, Bofeng 1; Tang, Feng 1; Huang, Wei 1,2
刊名	BIOORGANIC CHEMISTRY
	2020
卷号	94 页码:10
关键词	DSF Thermostability Glycosylation Glyco-engineered antibodies ADCs
ISSN号	0045-2068
DOI	10.1016/j.bioorg.2019.103391
通讯作者	Tang, Feng(tangfeng2013@simm.ac.cn) ; Huang, Wei(huangwei@simm.ac.cn)
英文摘要	Thermostability of monoclonal antibodies (mAbs) and antibody-drug conjugates (ADCs), as a critical property of biotherapeutics, is important for their physicochemical processes, pharmacodynamics, and pharmacokinetics. Fc glycosylation of mAbs plays a crucial role in antibody functions including thermostability, however, due to the lack of homogeneous glycosylation for comparison, the precise impact of glycoforms on thermostability of mAbs and ADCs remains challenging to elucidate. In this paper, we employed the technique of differential scanning fluorimetry (DSF) to investigate the thermostability of Fc domains, glycoengineered mAbs, and ADCs, carrying well-defined N-glycan structures for comparison. The results revealed that high-mannose-type N-glycans dramatically reduce the T-m value of Fc, compared to complex-type N-glycans. We also found that core-fucose contributes to the thermostability of mAbs, and the unnatural modification on non-reducing end of biantennary N-glycan can compensate the reduced stability of afucosylated mAbs and maintain the advantage of enhanced antibody-dependent cell-mediated cytotoxicity (ADCC). DSF analysis of lysine-linked and glycosite-specific ADCs indicated that thermostability of glycan-linked ADCs is reduced, but it could be improved by using an optimized linkage. This work provides an in-depth analysis on thermostability of mAbs and ADCs with homogeneous glycoforms, and also proposes new strategies for optimizing glycoengineered mAbs and glycosite-specific ADCs using unnatural glycan and stabilized linkage.
资助项目	National Natural Science Foundation of China (NNSFC)[21572244] ; National Natural Science Foundation of China (NNSFC)[21877116] ; National Science & Technology Major Project Key New Drug Creation and Manufacturing Program of China[2018ZX09711002-006] ; China Postdoctoral Science Foundation Postdoctoral Innovative Talent Support Program[BX20180339]
WOS关键词	THERMAL-STABILITY ; MONOCLONAL-ANTIBODY ; IGG ANTIBODIES ; GLYCOSYLATION ; AGGREGATION ; GLYCOFORM ; GLYCANS
WOS研究方向	Biochemistry & Molecular Biology ; Chemistry
语种	英语
出版者	ACADEMIC PRESS INC ELSEVIER SCIENCE
WOS记录号	WOS:000505596300047
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/282282]
专题	中国科学院上海药物研究所
通讯作者	Tang, Feng; Huang, Wei
作者单位	1.Chinese Acad Sci, CAS Ctr Excellence Mol Cell Sci, CAS Key Lab Receptor Res, Ctr Biotherapeut Discovery Res,Shanghai Inst Mat, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China 2.Univ Chinese Acad Sci, 19A Yuquan Rd, Beijing 100049, Peoples R China
推荐引用方式 GB/T 7714	Qin, Ken,Shi, Wei,Zhao, Lei,et al. Thermostability detection and optimization of glycoengineered antibodies and antibody-drug conjugates based on differential scanning flouremitry analysis[J]. BIOORGANIC CHEMISTRY,2020,94:10.
APA	Qin, Ken.,Shi, Wei.,Zhao, Lei.,Li, Mingjie.,Tang, Yubo.,...&Huang, Wei.(2020).Thermostability detection and optimization of glycoengineered antibodies and antibody-drug conjugates based on differential scanning flouremitry analysis.BIOORGANIC CHEMISTRY,94,10.
MLA	Qin, Ken,et al."Thermostability detection and optimization of glycoengineered antibodies and antibody-drug conjugates based on differential scanning flouremitry analysis".BIOORGANIC CHEMISTRY 94(2020):10.