Abstract:
:Spontaneous isopeptide bond formation, a stabilizing posttranslational modification that can be found in gram-positive bacterial cell surface proteins, has previously been used to develop a peptide-peptide ligation technology that enables the polymerization of tagged-proteins catalyzed by SpyLigase. Here we adapted this technology to establish a novel modular antibody labeling approach which is based on isopeptide bond formation between two recognition peptides, SpyTag and KTag. Our labeling strategy allows the attachment of a reporting cargo of interest to an antibody scaffold by fusing it chemically to KTag, available via semi-automated solid-phase peptide synthesis (SPPS), while equipping the antibody with SpyTag. This strategy was successfully used to engineer site-specific antibody-drug conjugates (ADCs) that exhibit cytotoxicities in the subnanomolar range. Our approach may lead to a new class of antibody conjugates based on peptide-tags that have minimal effects on protein structure and function, thus expanding the toolbox of site-specific antibody conjugation.
journal_name
Sci Repjournal_title
Scientific reportsauthors
Siegmund V,Piater B,Zakeri B,Eichhorn T,Fischer F,Deutsch C,Becker S,Toleikis L,Hock B,Betz UA,Kolmar Hdoi
10.1038/srep39291subject
Has Abstractpub_date
2016-12-16 00:00:00pages
39291issn
2045-2322pii
srep39291journal_volume
6pub_type
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