Proteome Profiling of 1 to 5 Spiked Circulating Tumor Cells Isolated from Whole Blood Using Immunodensity Enrichment, Laser Capture Microdissection, Nanodroplet Sample Processing, and Ultrasensitive nanoLC–MS
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Proteome profiling of circulating tumor cells (CTCs) can provide crucial insight into disease progression and the role of CTCs in tumor metastasis. We describe an integrated workflow to measure global protein expression in 1-5 spiked CTCs enriched from whole blood by immunodensity gradient centrifugation. Enriched CTCs were purified and collected by laser capture microdissection, prepared using a recently developed nanodroplet-based processing platform (nanoPOTS), and finally analyzed by ultrasensitive nanoLC-MS/MS. The workflow was capable of identifying an average of 164 and 607 protein groups from samples comprising 1 and 5 LNCaP cells, respectively, that were isolated from human whole blood. A panel of prostate cancer-specific proteins were identified and quantified, which was used to differentiate between spiked CTCs and white blood cells.
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Combining nanoPOTS with FACS and LCM has enabled single-cell proteomics and spatially resolved proteomics tissue mapping.
“By combining nanoPOTS with fluorescence-activated cell sorting (FACS) [26][27][28] and laser capture microdissection (LCM), 29,30 single cell proteomics and spatially resolved proteomics tissue mapping have been demonstrated.”
From Sensitive Top-Down Proteomics Analysis of a Low Number of Mammalian Cells Using a Nanodroplet Sample Processing Platform (2020) · cites this paper as [29]
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- Authors
- Zhu, Y., Podolak, J., Zhao, R., Shukla, A. K., Moore, R. J., Thomas, G. V., & Kelly, R. T.
- Affiliations
- Pacific Northwest National Laboratory (US)
- Oregon Health & Science University (US)
- Brigham Young University (US)
- Corresponding author
- Thomas, George V.
- Kelly, Ryan T.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2018-09-30
- Volume
- 90
- Issue
- 20
- Pages
- 11756-11759
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- PMC
- PMC6686195
- OpenAlex
- W2893216910
- Cited by
- 75
- References
- 23
- Citations by year
- 2026: 2 · 2025: 4 · 2024: 12 · 2023: 12 · 2022: 8 · 2021: 9 · 2020: 17 · 2019: 10 · 2018: 1
- FWCI
- 3.99
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Analytical Chemistry (publishedVersion)
- PubMed (publishedVersion)
- PubMed Central (acceptedVersion)
- Funders
- National Cancer Institute (P50 CA097186, R33 CA225248)
- Oregon Health and Science University
- National Institute of Biomedical Imaging and Bioengineering (R21 EB020976)
- Laboratory Directed Research and Development
- Knight Cancer Institute, Oregon Health and Science University
- Pacific Northwest National Laboratory
- Topics
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Metabolomics and Mass Spectrometry Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer Genomics and Diagnostics
- Keywords
- circulating tumor cells, proteomic profiling, laser capture microdissection, nanoLC-MS/MS, prostate cancer, LNCaP cells, tumor-specific proteins
- MeSH terms
- Centrifugation, Chromatography, Liquid, Humans, Immunohistochemistry, Neoplastic Cells, Circulating, Neoplasm Proteins, Particle Size, Proteome, Nanotechnology, Cell Line, Tumor, Tandem Mass Spectrometry, Nanoparticles, Laser Capture Microdissection
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