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

Ying Zhu; Jennifer Podolak; Rui Zhao; Anil K. Shukla; Ronald J. Moore; George V. Thomas; Ryan T. Kelly

doi:10.1021/acs.analchem.8b03268

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

Ying Zhu, Jennifer Podolak, Rui Zhao, Anil K. Shukla, Ronald J. Moore, George V. Thomas, Ryan T. Kelly

Knight Cancer Institute

Research output: Contribution to journal › Article › peer-review

25 Scopus citations

Abstract

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.

Original language	English (US)
Pages (from-to)	11756-11759
Number of pages	4
Journal	Analytical Chemistry
Volume	90
Issue number	20
DOIs	https://doi.org/10.1021/acs.analchem.8b03268
State	Published - Oct 16 2018

ASJC Scopus subject areas

Analytical Chemistry

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10.1021/acs.analchem.8b03268

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Zhu, Y., Podolak, J., Zhao, R., Shukla, A. K., Moore, R. J., Thomas, G. V., & Kelly, R. T. (2018). 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. Analytical Chemistry, 90(20), 11756-11759. https://doi.org/10.1021/acs.analchem.8b03268

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. / Zhu, Ying; Podolak, Jennifer; Zhao, Rui; Shukla, Anil K.; Moore, Ronald J.; Thomas, George V.; Kelly, Ryan T.

In: Analytical Chemistry, Vol. 90, No. 20, 16.10.2018, p. 11756-11759.

Research output: Contribution to journal › Article › peer-review

Zhu, Y, Podolak, J, Zhao, R, Shukla, AK, Moore, RJ, Thomas, GV & Kelly, RT 2018, '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', Analytical Chemistry, vol. 90, no. 20, pp. 11756-11759. https://doi.org/10.1021/acs.analchem.8b03268

Zhu Y, Podolak J, Zhao R, Shukla AK, Moore RJ, Thomas GV et al. 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. Analytical Chemistry. 2018 Oct 16;90(20):11756-11759. https://doi.org/10.1021/acs.analchem.8b03268

Zhu, Ying ; Podolak, Jennifer ; Zhao, Rui ; Shukla, Anil K. ; Moore, Ronald J. ; Thomas, George V. ; Kelly, Ryan T. / 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. In: Analytical Chemistry. 2018 ; Vol. 90, No. 20. pp. 11756-11759.

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title = "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",

abstract = "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.",

author = "Ying Zhu and Jennifer Podolak and Rui Zhao and Shukla, {Anil K.} and Moore, {Ronald J.} and Thomas, {George V.} and Kelly, {Ryan T.}",

note = "Funding Information: This work was supported by NIH Grants R33 CA225248 (R.T.K.) and R21 EB020976 (R.T.K.), the Pacific Northwest Prostate Cancer Grant SPORE/NCI P50 CA097186 (G.V.T), the OHSU-Knight Cancer Institute (G.V.T), Earth & Biological Sciences Directorate Mission Seed under the Laboratory Directed Research and Development Program at PNNL (Y.Z.), and the Precision Medicine Innovation Co-Laboratory (PMedIC), a joint research collaboration of OHSU and PNNL. A portion of this research was performed using EMSL, a national scientific user facility sponsored by the Department of Energy{\textquoteright}s Office of Biological and Environmental Research and located at PNNL.",

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AU - Thomas, George V.

AU - Kelly, Ryan T.

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