用于蛋白质工程的酵母表面展示：文库构建、筛选及亲和力成熟
Yeast Surface Display for Protein Engineering: Library Generation, Screening, and Affinity Maturation.
作者信息
Kang Byong H, Lax Brianna M, Wittrup K Dane
机构信息
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, USA.
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA, USA.
出版信息
Methods Mol Biol. 2022;2491:29-62. doi: 10.1007/978-1-0716-2285-8_2.
Yeast surface display is a powerful directed evolution method for developing and engineering protein molecules to attain desired properties. Here, updated protocols are presented for purposes of identification of lead binders and their affinity maturation. Large libraries are screened by magnetic bead selections followed by flow cytometric selections. Upon identification and characterization of single clones, their affinities are improved by an iterative process of mutagenesis and fluorescence-activated cell sorting.
酵母表面展示是一种强大的定向进化方法，用于开发和改造蛋白质分子以获得所需特性。本文介绍了用于鉴定先导结合物及其亲和力成熟的更新方案。通过磁珠筛选，随后进行流式细胞术筛选，对大型文库进行筛选。在鉴定和表征单个克隆后，通过诱变和荧光激活细胞分选的迭代过程提高它们的亲和力。
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