School of Molecular and Cellular Biology, Faculty of Biological Science, University of Leeds, Leeds LS2 9JT, UK.
BioScreening Technology Group, Faculty of Biological Science, University of Leeds, Leeds LS2 9JT, UK.
Sci Signal. 2017 Nov 14;10(505):eaan0868. doi: 10.1126/scisignal.aan0868.
Some 30 years after its discovery, phage display remains one of the most widely used methods of in vitro selection. Initially developed to revolutionize the generation of therapeutic antibodies, phage display is now the first choice for screening artificial binding proteins. Artificial binding proteins can be used as reagents to study protein-protein interactions, target posttranslational modifications, and distinguish between homologous proteins. They can also be used as research and affinity reagents, for diagnostic purposes, and as therapeutics. However, the ability to identify isoform-specific reagents remains highly challenging. We describe an adapted phage display protocol using an artificial binding protein (Affimer) for the selection of isoform-selective binding proteins.
噬菌体展示技术发现至今约 30 年，仍是体外筛选中应用最广泛的方法之一。噬菌体展示技术最初是为了彻底改变治疗性抗体的产生而开发的，如今已成为筛选人工结合蛋白的首选方法。人工结合蛋白可用作研究蛋白-蛋白相互作用、靶向翻译后修饰以及区分同源蛋白的试剂。它们还可以用作研究和亲和试剂、用于诊断目的以及作为治疗药物。然而，鉴定同工型特异性试剂的能力仍然极具挑战性。我们描述了一种经过改良的噬菌体展示技术方案，使用人工结合蛋白（Affimer）来筛选同工型选择性结合蛋白。
