Yeast Display for the Identification of Peptide-MHC Ligands of Immune Receptors.
Yeast Display for the Identification of Peptide-MHC Ligands of Immune Receptors.
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用于鉴定免疫受体肽-MHC 配体的酵母展示。
DOI：
10.1007/978-1-0716-2285-8_15
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发表时间：
2022
期刊：
影响因子：
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通讯作者：
Birnbaum,MichaelE
中科院分区：
文献类型：
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作者：
Huisman,BrookeD;Grace,BethE;Holec,PatrickV;Birnbaum,MichaelE
T细胞通过其T细胞受体（TCR）检测主要组织相容性复合体（MHC）蛋白呈递的肽抗原。可能的抗原序列多样性，具有数万亿潜在的肽- mhc靶点，使得研究，表征和操作给定TCR的肽库具有挑战性。酵母展示已被用于研究肽- mhc与T细胞受体之间的相互作用，以促进肽- mhc文库的高通量筛选。在这里，我们介绍了设计和验证肽- mhc酵母展示结构，设计和构建肽库，进行选择，准备，处理和分析肽库测序数据的见解。该方法的应用范围很广，包括表征TCR的肽- mhc识别谱，筛选已知TCR结合肽的高亲和力模位，以及从扩增的T细胞中鉴定TCR的天然配体。
T cells detect peptide antigens presented by major histocompatibility complex (MHC) proteins via their T cell receptor (TCR). The sequence diversity of possible antigens, with trillions of potential peptide-MHC targets, makes it challenging to study, characterize, and manipulate the peptide repertoire of a given TCR. Yeast display has been utilized to study the interactions between peptide-MHCs and T cell receptors to facilitate high-throughput screening of peptide-MHC libraries. Here we present insights on designing and validating a peptide-MHC yeast display construct, designing and constructing peptide libraries, conducting selections, and preparing, processing, and analyzing peptide library sequencing data. Applications for this approach are broad, including characterizing peptide-MHC recognition profiles for a TCR, screening for high-affinity mimotopes of known TCR-binding peptides, and identifying natural ligands of TCRs from expanded T cells.