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

Autor: Harmand TJ; Program in Cellular and Molecular Medicine , Boston Children's Hospital, Harvard Medical School , Boston , Massachussets 02115 , United States., Bousbaine D; Program in Cellular and Molecular Medicine , Boston Children's Hospital, Harvard Medical School , Boston , Massachussets 02115 , United States.; Department of Microbiology , Massachusetts Institute of Technology , Cambridge , Massachussets 02139 , United States., Chan A; The Broad Institute of Harvard and MIT, Howard Hughes Medical Institute, and the Department of Chemistry and Chemical Biology , Harvard University , Cambridge , Massachussets 02142 , United States., Zhang X; School of Biological Sciences , Nanyang Technological University , 637551 , Singapore., Liu DR; The Broad Institute of Harvard and MIT, Howard Hughes Medical Institute, and the Department of Chemistry and Chemical Biology , Harvard University , Cambridge , Massachussets 02142 , United States., Tam JP; School of Biological Sciences , Nanyang Technological University , 637551 , Singapore., Ploegh HL; Program in Cellular and Molecular Medicine , Boston Children's Hospital, Harvard Medical School , Boston , Massachussets 02115 , United States.
Jazyk: angličtina
Zdroj: Bioconjugate chemistry [Bioconjug Chem] 2018 Oct 17; Vol. 29 (10), pp. 3245-3249. Date of Electronic Publication: 2018 Sep 21.
DOI: 10.1021/acs.bioconjchem.8b00563
Abstrakt: 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.
Databáze: MEDLINE
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