抄録
Development of protein therapeutics or biosensors often requires in vitro affinity maturation. Here we report a robust affinity engineering strategy using a custom designed library. The strategy consists of two steps beginning with identification of beneficial single amino acid substitutions then combination. A high quality combinatorial library specifically customized to a given binding-interface can be rapidly designed by high-throughput mutational scanning of single substitution libraries. When applied to the optimization of a model antibody Fab fragment, the strategy created a diverse panel of high affinity variants. The most potent variant achieved 2110-fold affinity improvement to an equilibrium dissociation constant (Kd) of 3.45. pM with only 7 amino acid substitutions. The method should facilitate affinity engineering of a wide variety of protein-protein interactions due to its context-dependent library design strategy.
本文言語 | English |
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ページ(範囲) | 395-400 |
ページ数 | 6 |
ジャーナル | Biochemical and Biophysical Research Communications |
巻 | 428 |
号 | 3 |
DOI | |
出版ステータス | Published - 2012 11月 23 |
外部発表 | はい |
ASJC Scopus subject areas
- 生物理学
- 生化学
- 分子生物学
- 細胞生物学