Major histocompatibility complex (MHC)-presented peptides (pMHC) give insight into T cell immune responses, a critical step toward developing a new generation of targeted immunotherapies. Recent instrumentation advances have propelled mass spectrometry to being arguably the most robust technology for discovering and quantifying naturally presented pMHC from cells and tissues. However, sample preparation has remained a major limitation due to time-consuming and labor-intensive workflows. We developed a high-throughput and automated platform with enhanced speed, sensitivity, and reproducibility relative to prior studies. This pipeline is capable of processing up to 96 samples in 6 h or less yielding high-quality pMHC mixtures ready for mass spectrometry. Here, we describe our efforts to optimize purification and mass spectrometer parameters, ultimately allowing us to identify as many as almost 5000 pMHC I and 7400 pMHC II from as little as 2.5 × 10⁷ Raji cells each. We believe that this platform will facilitate and accelerate immunopeptidome profiling and benefit clinical research for immunotherapies.

中文翻译：

---

自动化的配体纯化平台通过质谱加速免疫肽组分析

主要组织相容性复合物（MHC）呈递的肽（pMHC）可以洞悉T细胞免疫反应，这是开发新一代靶向免疫疗法的关键一步。最近的仪器技术进步推动了质谱分析技术的发展，可以说它是从细胞和组织中发现和定量天然存在的pMHC的最强大技术。然而，由于耗时且劳动密集的工作流程，样品制备仍是主要限制。与以前的研究相比，我们开发了具有更高速度，灵敏度和可重复性的高通量自动化平台。该管道能够在6小时或更短的时间内处理多达96个样品，从而产生可用于质谱分析的高质量pMHC混合物。在这里，我们描述了我们为优化纯化和质谱仪参数所做的努力，每个^7个Raji细胞。我们相信，该平台将促进和加速免疫肽组分析，并有益于免疫疗法的临床研究。