One-step immobilization and purification of genetic engineering CBD fusion EndoS on cellulose for antibodies Fc-glycan remodeling

doi:10.1016/j.bioorg.2019.103114

	One-step immobilization and purification of genetic engineering CBD fusion EndoS on cellulose for antibodies Fc-glycan remodeling
	Zhao, Kai 1; Tang, Feng 2; Shi, Wei 2; Hong, Haofei 1; Zhou, Zhifang 1; Huang, Wei 2; Wu, Zhimeng 1
刊名	BIOORGANIC CHEMISTRY
	2019-10-01
卷号	91 页码:8
关键词	Endoglycosidase Cellulose binding domain (CBD) Cellulose Immobilization Antibody glycan remodeling
ISSN号	0045-2068
DOI	10.1016/j.bioorg.2019.103114
通讯作者	Tang, Feng(sztf05@126.com) ; Wu, Zhimeng(zwu@jiangnan.edu.cn)
英文摘要	The endoglycosidase (EndoS and its glycosynthase mutants D233A, D233Q) gene was fused with cellulose binding domain (CBD) using pET-35b vector and the fusion enzymes were successfully expressed in Escherichia coli. Then a simplified approach for one-step immobilization and purification of EndoS enzymes using cellulose as matrices were developed and excellent loading efficiency (81-90%) was achieved in optimal condition. The cellulose immobilized CBD-EndoS and the glycosynthase mutants presented high catalytic activity and were successfully applied in a two-step antibody Fc N-glycan remodeling, generating a therapeutic antibody with homogeneous glycoform in high efficiency. The cellulose immobilized CBD-EndoS and its mutants (D233A and D233Q) displayed excellent storage stability when stored at 4 degrees for one month. Reusability studies demonstrated that the cellulose immobilized CBD-EndoS and its mutants could be recycled for five times without obvious activity loss.
资助项目	National Natural Science Foundation of China (NNSFC)[21472070] ; National Natural Science Foundation of China (NNSFC)[21572244] ; China Postdoctoral Science Foundation[BX20180339] ; Priority Academic Program Development of Jiangsu Higher Education Institutions ; 111 Project[111-2-06] ; Open Foundation of Key Laboratory of Carbohydrate Chemistry & Biotechnology Ministry of Education[KLCCB-KF201804] ; Jiangsu province Collaborative Innovation Center for Advanced Industrial Fermentation industry development program
WOS关键词	STREPTOCOCCUS-PYOGENES ; BINDING PROPERTIES ; ENDOGLYCOSIDASE ; DOMAIN ; IGG ; EXPRESSION ; PROTEIN
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:000487812000013
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/288555]
专题	中国科学院上海药物研究所
通讯作者	Tang, Feng; Wu, Zhimeng
作者单位	1.Jiangnan Univ, Sch Biotechnol, Key Lab Carbohydrate Chem & Biotechnol, Minist Educ, 1800 Lihu Rd, Wuxi 214122, Jiangsu, Peoples R China 2.Chinese Acad Sci, CAS Ctr Excellence Mol Cell Sci, Shanghai Inst Mat Med, CAS Key Lab Receptor Res, 555 Zuchongzhi Rd, Shanghai 201203, Peoples R China
推荐引用方式 GB/T 7714	Zhao, Kai,Tang, Feng,Shi, Wei,et al. One-step immobilization and purification of genetic engineering CBD fusion EndoS on cellulose for antibodies Fc-glycan remodeling[J]. BIOORGANIC CHEMISTRY,2019,91:8.
APA	Zhao, Kai.,Tang, Feng.,Shi, Wei.,Hong, Haofei.,Zhou, Zhifang.,...&Wu, Zhimeng.(2019).One-step immobilization and purification of genetic engineering CBD fusion EndoS on cellulose for antibodies Fc-glycan remodeling.BIOORGANIC CHEMISTRY,91,8.
MLA	Zhao, Kai,et al."One-step immobilization and purification of genetic engineering CBD fusion EndoS on cellulose for antibodies Fc-glycan remodeling".BIOORGANIC CHEMISTRY 91(2019):8.