Kalaora Shelly, Samuels Yardena
Molecular Cell Biology Department, Weizmann Institute of Science, Rehovot, Israel.
Methods Mol Biol. 2019;1884:203-214. doi: 10.1007/978-1-4939-8885-3_14.
Neo-antigens expressed on tumors are targets for development of cancer immunotherapy strategies. Use of prediction algorithms to identify neo-antigens yields a significant number of peptides that must be validated in laborious and time-consuming methods; many prove to be false-positive identifications. The use of HLA peptidomics allows the isolation of the HLA-peptide complexes directly from cells and can be done on fresh tumor, patient-derived xerographs, or cell lines when the tissue sample is limited. This method can be used to identify both HLA class I and HLA class II or any different MHC from different species. Here we describe the steps to create the immune-affinity columns used from the process, the immunoprecipitation procedure, and also the isolation of the peptides that will be analyzed by mass spectrometry.
肿瘤上表达的新抗原是癌症免疫治疗策略开发的靶点。使用预测算法来识别新抗原会产生大量肽段，这些肽段必须通过费力且耗时的方法进行验证；许多结果被证明是假阳性识别。HLA肽组学的应用允许直接从细胞中分离HLA-肽复合物，并且当组织样本有限时，可以在新鲜肿瘤、患者来源的干片或细胞系上进行。该方法可用于识别HLA I类和HLA II类，或来自不同物种的任何不同的MHC。在这里，我们描述了从该过程中创建免疫亲和柱的步骤、免疫沉淀程序，以及将通过质谱分析的肽段的分离。
