One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1

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 shor...

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Main Authors:	Harmand, Thibault J., Bousbaine, Djenet, Chan, Alix, Zhang, Xiaohong, Liu, David R., Tam, James P., Ploegh, Hidde L.
Other Authors:	School of Biological Sciences
Format:	Article
Language:	English
Published:	2020
Subjects:	Science::Biological sciences::Biochemistry Immunology Peptides and Proteins
Online Access:	https://hdl.handle.net/10356/143935
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Institution:	Nanyang Technological University
Language:	English

id	sg-ntu-dr.10356-143935
record_format	dspace
spelling	sg-ntu-dr.10356-1439352020-10-02T02:05:11Z One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1 Harmand, Thibault J. Bousbaine, Djenet Chan, Alix Zhang, Xiaohong Liu, David R. Tam, James P. Ploegh, Hidde L. School of Biological Sciences Science::Biological sciences::Biochemistry Immunology Peptides and Proteins 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. 2020-10-02T02:05:11Z 2020-10-02T02:05:11Z 2018 Journal Article Harmand, T. J., Bousbaine, D., Chan, A., Zhang, X., Liu, D. R., Tam, J. P., & Ploegh, H. L. (2018). One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1. Bioconjugate Chemistry, 29(10), 3245-3249. doi:10.1021/acs.bioconjchem.8b00563 1520-4812 https://hdl.handle.net/10356/143935 10.1021/acs.bioconjchem.8b00563 30231608 10 29 3245 3249 en Bioconjugate chemistry © 2018 American Chemical Society. All rights reserved.
institution	Nanyang Technological University
building	NTU Library
country	Singapore
collection	DR-NTU
language	English
topic	Science::Biological sciences::Biochemistry Immunology Peptides and Proteins
spellingShingle	Science::Biological sciences::Biochemistry Immunology Peptides and Proteins Harmand, Thibault J. Bousbaine, Djenet Chan, Alix Zhang, Xiaohong Liu, David R. Tam, James P. Ploegh, Hidde L. One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1
description	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.
author2	School of Biological Sciences
author_facet	School of Biological Sciences Harmand, Thibault J. Bousbaine, Djenet Chan, Alix Zhang, Xiaohong Liu, David R. Tam, James P. Ploegh, Hidde L.
format	Article
author	Harmand, Thibault J. Bousbaine, Djenet Chan, Alix Zhang, Xiaohong Liu, David R. Tam, James P. Ploegh, Hidde L.
author_sort	Harmand, Thibault J.
title	One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1
title_short	One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1
title_full	One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1
title_fullStr	One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1
title_full_unstemmed	One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1
title_sort	one-pot dual labeling of igg 1 and preparation of c-to-c fusion proteins through a combination of sortase a and butelase 1
publishDate	2020
url	https://hdl.handle.net/10356/143935
_version_	1681057929417981952

One-pot dual labeling of IgG 1 and preparation of C-to-C fusion proteins through a combination of sortase A and butelase 1

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