Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE, USA.
Department of Chemical & Biomolecular Engineering, University of Nebraska-Lincoln, Lincoln, NE, USA.
Methods Mol Biol. 2023;2705:293-305. doi: 10.1007/978-1-0716-3393-9_16.
Protein engineering has brought advances to industrial processes, biomaterials, nanotechnology, biosensors, and biomedical applications. This chapter will focus on the engineering of Src Homology 2 domains (SH2) to act as an antibody mimetic for the recognition of sulfotyrosine-containing peptides or proteins. In comparison to anti-sulfotyrosine antibodies, SH2 mutants have much smaller size and can be heterologously expressed and purified in large quantity at low cost. This chapter will describe the use of phage display to identify a sulfotyrosine-binding SH2 mutant and the subsequent enrichment of sulfotyrosine-containing peptides in complex biological samples.
蛋白质工程在工业过程、生物材料、纳米技术、生物传感器和生物医学应用方面带来了进步。本章将重点介绍Src Homology 2 结构域（SH2）的工程改造，以作为模拟抗体来识别含硫酪氨酸的肽或蛋白质。与抗硫酪氨酸抗体相比，SH2 突变体的体积要小得多，可以异源表达并以低成本大量纯化。本章将描述使用噬菌体展示来鉴定结合硫酪氨酸的 SH2 突变体，以及随后在复杂生物样品中富集含硫酪氨酸的肽。
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