高防护包膜病毒感染的人细胞 HLA-I 免疫肽组分析。
HLA-I immunopeptidome profiling of human cells infected with high-containment enveloped viruses.
机构信息
Broad Institute of MIT and Harvard, Cambridge, MA, USA; Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, MA, USA; Laboratory of Virology and Infectious Disease, The Rockefeller University, New York, NY, USA.
Broad Institute of MIT and Harvard, Cambridge, MA, USA.
出版信息
STAR Protoc. 2022 Dec 16;3(4):101910. doi: 10.1016/j.xpro.2022.101910. Epub 2022 Dec 8.
Immunopeptidome profiling of infected cells is a powerful technique for detecting viral peptides that are naturally processed and loaded onto class I human leukocyte antigens (HLAs-I). Here, we provide a protocol for preparing samples for immunopeptidome profiling that can inactivate enveloped viruses while still preserving the integrity of the HLA-I complex. We detail steps for lysate preparation of infected cells followed by HLA-I immunoprecipitation and virus inactivation. We further describe peptide purification for mass spectrometry outside a high-containment facility. For complete details on the use and execution of this protocol, please refer to Weingarten-Gabbay et al. (2021)..
对感染细胞的免疫肽组进行分析是一种强大的技术，可以检测到经过自然加工并加载到 I 类人白细胞抗原（HLA-I）上的病毒肽。在这里，我们提供了一种用于免疫肽组分析样品制备的方案，该方案可以在灭活包膜病毒的同时保持 HLA-I 复合物的完整性。我们详细介绍了感染细胞裂解物制备、HLA-I 免疫沉淀和病毒灭活的步骤。我们进一步描述了在高防护设施外进行质谱分析的肽纯化步骤。有关此方案的使用和执行的完整详细信息，请参阅 Weingarten-Gabbay 等人（2021）。
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