Marino Fabio, Chong Chloe, Michaux Justine, Bassani-Sternberg Michal
Ludwig Centre for Cancer Research, University of Lausanne, Epalinges, Switzerland.
Department of Oncology, University Hospital of Lausanne (CHUV), Lausanne, Switzerland.
Methods Mol Biol. 2019;1913:67-79. doi: 10.1007/978-1-4939-8979-9_5.
Comprehensive knowledge of the HLA class I and class II peptides presented to T cells is crucial for designing innovative therapeutics against cancer and other diseases. So far, methodologies for recovery of HLA class I and II peptides for subsequent mass spectrometry-based analysis have been a major limitation. In this chapter we describe a detailed protocol for a high-throughput, reproducible, and sensitive immunoaffinity-purification of HLA-I and HLA-II peptides from up to 96 samples in a plate format, suitable for tissue samples and cell lines. Our methodology reduces sample handling, has a competitive peptide yield, and can be completed within 5 h. This simplified pipeline is applicable for basic and clinical applications.
全面了解呈递给T细胞的HLA I类和II类肽对于设计针对癌症和其他疾病的创新疗法至关重要。到目前为止，用于回收HLA I类和II类肽以便后续基于质谱分析的方法一直是一个主要限制。在本章中，我们描述了一种详细的方案，用于以板形式从多达96个样品中高通量、可重复且灵敏地免疫亲和纯化HLA-I和HLA-II肽，适用于组织样品和细胞系。我们的方法减少了样品处理，具有有竞争力的肽产量，并且可以在5小时内完成。这种简化的流程适用于基础和临床应用。
