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Abstract：
Covalent target modulation with small molecules has been emerging as a promising strategy for drug discovery. However, covalent inhibitory antibody remains unexplored due to the lack of efficient strategies to engineer antibody with desired bioactivity. Herein, we developed an intracellular selection method to generate covalent inhibitory antibody against human rhinovirus 14 (HRV14) 3C protease through unnatural amino acid mutagenesis along the heavy chain complementarity-determining region 3 (CDR-H3). A library of antibody mutants was thus constructed and screened in vivo through co-expression with the target protease. Using this screening strategy, six covalent antibodies with proximity-enabled bioactivity were identified, which were shown to covalently target HRV14-3C protease with high inhibitory potency and exquisite selectivity. Compared to structure-based rational design, this library-based screening method provides a simple and efficient way for the discovery and engineering of covalent antibody for enzyme inhibition.
摘要：
用小分子进行共价靶标调节已经成为一种有希望的药物发现策略。然而,共价抑制性抗体仍未被开发,因为缺乏有效的策略来工程化具有所需生物活性的抗体。在这里，我们开发了一种细胞内选择方法，通过沿着重链互补决定区3（CDR-H3）的非天然氨基酸诱变来产生针对人鼻病毒14（HRV14）3C蛋白酶的共价抑制性抗体。因此构建了抗体突变体的文库,并通过与靶蛋白酶共表达在体内筛选。使用这种筛查策略，鉴定了六种具有邻近使能生物活性的共价抗体，显示其共价靶向HRV14-3C蛋白酶，具有高抑制效力和精细选择性。与基于结构的合理设计相比，这种基于文库的筛选方法为酶抑制共价抗体的发现和工程化提供了一种简单有效的方法。
