Strategy for Microscale Extraction and Proteome Profiling of Peripheral Blood Mononuclear Cells.
Anal Chem
; 94(25): 8827-8832, 2022 06 28.
Article
in English
| MEDLINE | ID: covidwho-1890086
ABSTRACT
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 omics 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 µL 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 µL 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-κB 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.
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Leukocytes, Mononuclear
/
COVID-19
Type of study:
Prognostic study
Topics:
Vaccines
Limits:
Female
/
Humans
/
Male
Language:
English
Journal:
Anal Chem
Year:
2022
Document Type:
Article
Affiliation country:
Acs.analchem.1c05365
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