Synthesis of epitope-targeting nanobody based on native protein–protein interactions for FtsZ filamentation suppressor

Hikaru Nakazawa, Taiji Katsuki, Takashi Matsui, Atsushi Tsugita, Takeshi Yokoyama, Tomoyuki Ito, Sakiya Kawada, Yoshikazu Tanaka, Mitsuo Umetsu

研究成果: ジャーナルへの寄稿学術論文査読

抄録

Phage display and biopanning are powerful tools for generating binding molecules for a specific target. However, the selection process based only on binding affinity provides no assurance for the antibody's affinity to the target epitope. In this study, we propose a molecular-evolution approach guided by native protein–protein interactions to generate epitope-targeting antibodies. The binding-site sequence in a native protein was grafted into a complementarity-determining region (CDR) in the nanobody, and a nonrelated CDR loop (in the grafted nanobody) was randomized to create a phage display library. In this construction of nanobodies by integrating graft and evolution technology (CAnIGET method), suitable grafting of the functional sequence added functionality to the nanobody, and the molecular-evolution approach enhanced the binding function to inhibit the native protein–protein interactions. To apply for biological tool with growth screening, model nanobodies with an affinity for filamenting temperature-sensitive mutant Z (FtsZ) from Staphylococcus aureus were constructed and completely inhibited the polymerization of FtsZ as a function. Consequently, the expression of these nanobodies drastically decreased the cell division rate. We demonstrate the potential of the CAnIGET method with the use of native protein–protein interactions for steady epitope-specific evolutionary engineering.

本文言語英語
論文番号2300039
ジャーナルBiotechnology Journal
18
11
DOI
出版ステータス出版済み - 2023 11月

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