Incorporation of Halogenated Amino Acids into Antibody Fragments at Multiple Specific Sites Enhances Antigen Binding.
Fab
Rituximab
bromotyrosine
shape complementarity
unnatural amino acid incorporation
Journal
Chembiochem : a European journal of chemical biology
ISSN: 1439-7633
Titre abrégé: Chembiochem
Pays: Germany
ID NLM: 100937360
Informations de publication
Date de publication:
05 01 2021
05 01 2021
Historique:
received:
01
07
2020
revised:
08
08
2020
pubmed:
21
8
2020
medline:
24
9
2021
entrez:
21
8
2020
Statut:
ppublish
Résumé
Expansion of the amino-acid repertoire with synthetic derivatives introduces novel structures and functionalities into proteins. In this study, we improved the antigen binding of antibodies by incorporating halogenated tyrosines at multiple selective sites. Tyrosines in the Fab fragment of an anti-EGF-receptor antibody 059-152 were systematically replaced with 3-bromo- and 3-chlorotyrosines, and simultaneous replacements at four specific sites were found to cause a tenfold increase in the affinity toward the antigen. Structure modeling suggested that this effect was due to enhanced shape complementarity between the antigen and antibody molecules. On the other hand, we showed that chlorination in the constant domain, far from the binding interface, of Rituximab Fab also increased the affinity significantly (up to 17-fold). Our results showed that antigen binding is tunable with the halogenation in and out of the binding motifs.
Identifiants
pubmed: 32815262
doi: 10.1002/cbic.202000429
doi:
Substances chimiques
Amino Acids
0
Antibodies, Monoclonal
0
Antigens
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
120-123Informations de copyright
© 2020 Wiley-VCH GmbH.
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