Institute of Applied Biosciences, Centre for Research and Technology Hellas, Thessaloniki, Greece.
Department of Molecular Biology and Genetics (MBG), Democritus University of Thrace, Alexandroupolis, Greece.
Methods Mol Biol. 2022;2453:585-603. doi: 10.1007/978-1-0716-2115-8_27.
The study of antigen receptor gene repertoires using next-generation sequencing (NGS) technologies has disclosed an unprecedented depth of complexity, requiring novel computational and analytical solutions. Several bioinformatics workflows have been developed to this end, including the T-cell receptor/immunoglobulin profiler (TRIP), a web application implemented in R shiny, specifically designed for the purposes of comprehensive repertoire analysis, which is the focus of this chapter. TRIP has the potential to perform robust immunoprofiling analysis through the extraction and processing of the IMGT/HighV-Quest output, via a series of functions, ensuring the analysis of high-quality, biologically relevant data through a multilevel process of data filtering. Subsequently, it provides in-depth analysis of antigen receptor gene rearrangements, including (a) clonality assessment; (b) extraction of variable (V), diversity (D), and joining (J) gene repertoires; (c) CDR3 characterization at both the nucleotide and amino acid level; and (d) somatic hypermutation analysis, in the case of immunoglobulin gene rearrangements. Relevant to mention, TRIP enables a high level of customization through the integration of various options in key aspects of the analysis, such as clonotype definition and computation, hence allowing for flexibility without compromising on accuracy.
使用下一代测序 (NGS) 技术研究抗原受体基因库,揭示了前所未有的复杂性,这需要新的计算和分析解决方案。为此已经开发了几种生物信息学工作流程,包括 T 细胞受体/免疫球蛋白分析器 (TRIP),这是一个在 R shiny 中实现的 Web 应用程序,专门设计用于全面的库分析,这是本章的重点。TRIP 具有通过提取和处理 IMGT/HighV-Quest 输出进行强大免疫分析的潜力,通过一系列功能确保通过多层次的数据过滤过程分析高质量、有生物学意义的数据。随后,它提供了对抗原受体基因重排的深入分析,包括 (a) 克隆性评估;(b) 提取可变 (V)、多样性 (D) 和连接 (J) 基因库;(c) 在核苷酸和氨基酸水平上对 CDR3 进行特征描述;以及 (d) 在免疫球蛋白基因重排的情况下进行体细胞超突变分析。值得一提的是,TRIP 通过在分析的关键方面集成各种选项(例如克隆型定义和计算),实现了高水平的定制化,从而在不影响准确性的情况下具有灵活性。