One-Pot Dual Labeling of IgG 1 and Preparation of C-to-C Fusion Proteins Through a Combination of Sortase A and Butelase 1

被引：73

作者：

Harmand, Thibault J. ^{[1
]}

Bousbaine, Djenet ^{[1
,2
]}

Chan, Alix ^{[3
,4
]}

Zhang, Xiaohong ^{[5
]}

Liu, David R. ^{[3
,4
]}

Tam, James P. ^{[5
]}

Ploegh, Hidde L. ^{[1
]}

机构：

[1] Harvard Med Sch, Program Cellular & Mol Med, Boston Childrens Hosp, Boston, MA 02115 USA

[2] MIT, Dept Microbiol, 77 Massachusetts Ave, Cambridge, MA 02139 USA

[3] Harvard Univ, Howard Hughes Med Inst, Brd Inst Harvard & MIT, Cambridge, MA 02142 USA

[4] Harvard Univ, Dept Chem & Chem Biol, Cambridge, MA 02142 USA

[5] Nanyang Technol Univ, Sch Biol Sci, Singapore 637551, Singapore

来源：

BIOCONJUGATE CHEMISTRY | 2018年 / 29卷 / 10期

基金：

瑞士国家科学基金会;

关键词：

ANTIBODY DRUG CONJUGATE; UNNATURAL AMINO-ACIDS; CHEMICAL-SYNTHESIS; FRET;

D O I：

10.1021/acs.bioconjchem.8b00563

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Site-specific chemical modification of proteins can assist in the study of their function. Furthermore, these methods are essential to develop biologicals for diagnostic and therapeutic use. Standard protein engineering protocols and recombinant expression enable the production of proteins with short peptide tags recognized by enzymes capable of site-specific modification. We report here the application of two enzymes of orthogonal specificity, sortase A and butelase 1, to prepare non-natural C-to-C fusion proteins. Using these enzymes, we further demonstrate site-selective installation of different chemical moieties at two sites in a full-size antibody molecule.

引用

页码：3245 / 3249

页数：5

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