酵母展示引导的 pH 依赖性配体的选择。
Yeast Display Guided Selection of pH-Dependent Binders.
机构信息
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC, USA.
Golden LEAF Biomanufacturing Training and Education Center (BTEC), North Carolina State University, Raleigh, NC, USA.
出版信息
Methods Mol Biol. 2022;2491:293-311. doi: 10.1007/978-1-0716-2285-8_16.
Abstract
pH-dependent antigen binding has proven useful in engineering next-generation therapeutics specifically via antibody recycling technology. This technology allows for half-life extension, thereby lowering the amount and frequency of dosing of therapeutics. Cell sorting, coupled with display techniques, has been used extensively for the selection of high-affinity binders. Herein, we describe a cell sorting methodology utilizing yeast surface display for selection of binding proteins with strong binding at physiological pH and weak to no binding at acidic pH. This methodology can be readily applied to engineer proteins and/or antibodies that do not have pH-dependent binding or for selection of de novo pH-dependent binders using library-based methods.
摘要
pH 依赖性抗原结合已被证明在工程下一代治疗剂方面非常有用，特别是通过抗体回收技术。该技术允许半衰期延长，从而降低治疗剂的剂量和频率。细胞分选与展示技术已广泛用于高亲和力结合物的选择。本文描述了一种利用酵母表面展示进行细胞分选的方法，用于选择在生理 pH 下具有强结合性而在酸性 pH 下弱结合或不结合的结合蛋白。该方法可以很容易地应用于工程化不具有 pH 依赖性结合的蛋白质和/或抗体，或者用于使用基于文库的方法选择新的 pH 依赖性结合物。
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