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タイトル: | Peptide barcoding for one-pot evaluation of sequence–function relationships of nanobodies |
著者: | Matsuzaki, Yusei Aoki, Wataru https://orcid.org/0000-0002-3118-9390 (unconfirmed) Miyazaki, Takumi Aburaya, Shunsuke Ohtani, Yuta Kajiwara, Kaho Koike, Naoki Minakuchi, Hiroyoshi Miura, Natsuko Kadonosono, Tetsuya Ueda, Mitsuyoshi |
著者名の別形: | 松崎, 友星 青木, 航 宮崎, 拓己 油屋, 駿介 大谷, 優太 梶原, 果穂 小池, 直暉 水口, 博義 三浦, 夏子 門之園, 哲哉 植田, 充美 |
キーワード: | Antibody therapy High-throughput screening Mass spectrometry |
発行日: | 2021 |
出版者: | Springer Nature |
誌名: | Scientific Reports |
巻: | 11 |
論文番号: | 21516 |
抄録: | Optimisation of protein binders relies on laborious screening processes. Investigation of sequence–function relationships of protein binders is particularly slow, since mutants are purified and evaluated individually. Here we developed peptide barcoding, a high-throughput approach for accurate investigation of sequence–function relationships of hundreds of protein binders at once. Our approach is based on combining the generation of a mutagenised nanobody library fused with unique peptide barcodes, the formation of nanobody–antigen complexes at different ratios, their fine fractionation by size-exclusion chromatography and quantification of peptide barcodes by targeted proteomics. Applying peptide barcoding to an anti-GFP nanobody as a model, we successfully identified residues important for the binding affinity of anti-GFP nanobody at once. Peptide barcoding discriminated subtle changes in KD at the order of nM to sub-nM. Therefore, peptide barcoding is a powerful tool for engineering protein binders, enabling reliable one-pot evaluation of sequence–function relationships. |
記述: | 遊離型抗体の構造活性相関解析を迅速に評価可能とする新手法を開発. 京都大学プレスリリース. 2021-11-08. |
著作権等: | © The Author(s) 2021 This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. |
URI: | http://hdl.handle.net/2433/265862 |
DOI(出版社版): | 10.1038/s41598-021-01019-6 |
PubMed ID: | 34728738 |
関連リンク: | https://www.kyoto-u.ac.jp/ja/research-news/2021-11-08-2 |
出現コレクション: | 学術雑誌掲載論文等 |
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