【Optimized Platform for In-Depth Characterization of MHC Class I and II Post-translationally Modified Immunopeptides】
Characterization of major histocompatibility complex (MHC) peptides provides key insight into the adaptive immune system but also presents a significant analytical challenge. Detecting rare, immunogenic peptides has long been a challenge for proteomics due to their low abundance, non-tryptic peptide properties, and the many steps required in enrichment. Here, we present an optimized platform allowing the characterization of broadly expressed MHC class I and class II immunopeptides as well as downstream enrichment for post- translational modifications (PTMs) such as phosphorylation. We achieved an enhanced depth of the MHC-I and II immunopeptidomes by optimizing multiple sample preparation strategies, DDA-PASEF mass spectrometry methods, and data analysis, while using less starting material compared to previously published approaches
【MHCBooster: An AI-powered Software to Boost DDA-based Immunopeptide Identification】
识别由主要组织相容性复合体(MHC)分子所呈现的免疫肽对于理解免疫反应以及开发基于 T 细胞的靶向疫苗和免疫疗法至关重要。然而,通过质谱(MS)对免疫肽进行精确识别,尤其是在从低上样量时,是一项极具挑战性的任务。为了解决这个问题,我们基于Bruker timsTOF质谱平台,开发了 MHCBooster 这一基于人工智能的工具,它利用深度学习模型来利用数据依赖式采集(DDA)质谱技术来增强免疫肽的识别能力。通过为 MHC-I 和 MHC-II 肽提供诸如保留时间(RT)、MS2、离子迁移率(IM)以及抗原加工和呈递(APP)等关键维度的可靠性测量值,MHCBooster 在表位识别方面提高了灵敏度和特异性,尤其是在低输入情况下。MHCBooster 具有图形用户界面(GUI),并且还可以通过命令行或作为 Python 包进行使用。