期刊
ANALYTICAL CHEMISTRY
卷 94, 期 25, 页码 8827-8832出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.1c05365
关键词
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资金
- National Key Research and Development Project of China [2021YFA1300200, 2021YFA130160]
- Research Program of the State Key Laboratory of Proteomics [SKLP-K201901]
Peripheral blood mononuclear cells (PBMCs) play vital roles in disease monitoring and physiological processes. Researchers have developed a microscale apparatus (PBMC-mCap) to enrich and analyze the proteome of PBMCs from small volumes of peripheral blood. This approach provides a convenient clinical toolkit for disease diagnosis and monitoring of the healthy state.
Peripheral blood mononuclear cells (PBMCs) play vital roles in physiological and pathological processes and represent a rich source for disease monitoring. Typical molecular profiling on PBMCs involves the sorting of cell subsets and thus requires a large volume of peripheral blood (PB), which impedes the clinical practicability of omits tools in PBMC measurements. It would be clinically invaluable to develop a convenient approach for preparing PBMCs from small volumes of PB and for deep proteome profiling of PBMCs. To this end, here, we designed an apparatus (PBMC-mCap) for microscale enrichment and proteome analysis of PBMCs, which pushed the needed PB volume from the normal 2 mL or higher to 100 mu L or lower, comparable to the volume of a drop of finger blood. A PBMC-specific mass spectra library containing 8869 proteins and 121,956 peptides was further built, which, in combination with the optimized data-independent acquisition strategy, helped to identify 6000 and 6500 proteins from PBMCs with 100 mu L and 1 mL of PB as initial materials, respectively. Further application of the strategy for PBMC proteomes revealed a steady difference between gender (male vs female) and upon stimulus (COVID-19 vaccination). For the latter, we observed differentially expressed genes and pathways involving the activation of immune cells, including the NF-kappa B pathway, inflammation response, and antiviral response. Our strategy for the proteome analysis of microscale PBMCs may provide a convenient clinical toolkit for disease diagnosis and healthy state monitoring.
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