High-Throughput Single-Cell Immunoassay in the Cellular Native Environment Using Online Desalting Dual-Spray Mass Spectrometry

Single-cell mass spectrometry (MS) remains challenging in the analysis of cells in the native environment due to the severe ion suspension from nonvolatile salts. Synchronous desalting and ionization would be ideal to both ensure the native environment and remove the salt interference. Here, a novel...

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Published inAnalytical chemistry (Washington) Vol. 92; no. 24; pp. 15854 - 15861
Main Authors Xu, Shuting, Xue, Jinjuan, Bai, Yu, Liu, Huwei
Format Journal Article
LanguageEnglish
Published United States American Chemical Society 15.12.2020
Subjects
Antigens
Biomarkers, Tumor - analysis
Buffers
CA-125 Antigen - analysis
Cancer
Cell surface
Chemistry
Desalination
Environments
High-Throughput Screening Assays
Humans
Immunoassay
Ionization
Ions
Lipids
Mass Spectrometry
Mass spectroscopy
Particle Size
Proteins
Rhodamine
Salts
Scientific imaging
Single-Cell Analysis
Spectroscopy
Surface Properties
Tags
Tumor cell lines
Tumor Cells, Cultured
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Summary:Single-cell mass spectrometry (MS) remains challenging in the analysis of cells in the native environment due to the severe ion suspension from nonvolatile salts. Synchronous desalting and ionization would be ideal to both ensure the native environment and remove the salt interference. Here, a novel dual-spray ionization technique combining electrospray and nanoelectrospray ionization (ESI-nESI) was developed, enabling highly efficient online desalting during the ionization process. In situ detection of cell surface proteins from the intact cells in phosphate buffer saline (PBS) was achieved by dual ESI-nESI MS with the help of an MS-based immunoassay using rhodamine-based mass tags. These mass tags were confirmed to be highly competitive during desalting, which improved the protein detection sensitivity to a single-cell level. Through the combination of the single-cell immunoassay with ESI-nESI MS, the important surface protein markers, cancer antigen 125, in two cancer cell lines (OVCAR-3 and MCF-7) suspended in the PBS buffers were screened in a high-throughput cytometric mode, along with some proposed cellular endogenous lipids. The ESI-nESI MS system is promising for multidimensional organic mass cytometric analysis in the cellular native environment for clinical use and many basic biology researches.
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ISSN:0003-2700
1520-6882
DOI:10.1021/acs.analchem.0c03167