Incorporation of Halogenated Amino Acids into Antibody Fragments at Multiple Specific Sites Enhances Antigen Binding
| Autor: | Haruna Sato, Kensaku Sakamoto, Akiko Hayashi, Takuhiro Ito, Kenichiro Ito, Haruna Kenichi, Chisato Makino |
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| Rok vydání: | 2020 |
| Předmět: |
Models
Molecular Halogenation Constant domain 010402 general chemistry 01 natural sciences Biochemistry Antibody fragments Antigen-Antibody Reactions Antigen Molecule Amino Acids Antigens Molecular Biology chemistry.chemical_classification Binding Sites biology 010405 organic chemistry Chemistry Organic Chemistry Antibodies Monoclonal Antigen binding 0104 chemical sciences Amino acid biology.protein Molecular Medicine Antibody |
| Zdroj: | Chembiochem : a European journal of chemical biology. 22(1) |
| ISSN: | 1439-7633 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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