Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
Shanghai Center for Systems Biomedicine, Key Laboratory of Systems Biomedicine (Ministry of Education), Shanghai Jiao Tong University, Shanghai, China.
Mol Cell Proteomics. 2024 Sep;23(9):100831. doi: 10.1016/j.mcpro.2024.100831. Epub 2024 Aug 19.
Characterizing the antibody reactome for circulating antibodies provide insight into pathogen exposure, allergies, and autoimmune diseases. This is important for biomarker discovery, clinical diagnosis, and prognosis of disease progression, as well as population-level insights into the immune system. The emerging technology phage display immunoprecipitation and sequencing (PhIP-seq) is a high-throughput method for identifying antigens/epitopes of the antibody reactome. In PhIP-seq, libraries with sequences of defined lengths and overlapping segments are bioinformatically designed using naturally occurring proteins and cloned into phage genomes to be displayed on the surface. These libraries are used in immunoprecipitation experiments of circulating antibodies. This can be done with parallel samples from multiple sources, and the DNA inserts from the bound phages are barcoded and subjected to next-generation sequencing for hit determination. PhIP-seq is a powerful technique for characterizing the antibody reactome that has undergone rapid advances in recent years. In this review, we comprehensively describe the history of PhIP-seq and discuss recent advances in library design and applications.
描述循环抗体的抗体反应组可以深入了解病原体暴露、过敏和自身免疫性疾病。这对于生物标志物的发现、临床诊断和疾病进展的预后，以及对免疫系统的人群水平的了解都很重要。新兴的噬菌体展示免疫沉淀和测序（PhIP-seq）技术是一种高通量的方法，用于鉴定抗体反应组的抗原/表位。在 PhIP-seq 中，使用天然存在的蛋白质设计具有定义长度和重叠片段的文库，并将其克隆到噬菌体基因组中，以在表面展示。这些文库用于循环抗体的免疫沉淀实验。这可以在来自多个来源的平行样本中进行，并且结合噬菌体的 DNA 插入物进行条形码标记，并进行下一代测序以确定命中。PhIP-seq 是一种强大的技术，可以描述抗体反应组，近年来取得了快速进展。在这篇综述中，我们全面描述了 PhIP-seq 的历史，并讨论了文库设计和应用的最新进展。
