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 | |
| State | Published - Oct 16 2018 |
Fingerprint
ASJC Scopus subject areas
- Analytical Chemistry
Cite this
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; Kelly, Ryan T.
In: Analytical Chemistry, Vol. 90, No. 20, 16.10.2018, p. 11756-11759.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - 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
AU - Zhu, Ying
AU - Podolak, Jennifer
AU - Zhao, Rui
AU - Shukla, Anil K.
AU - Moore, Ronald J.
AU - Thomas, George
AU - Kelly, Ryan T.
PY - 2018/10/16
Y1 - 2018/10/16
N2 - 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.
AB - 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.
UR - http://www.scopus.com/inward/record.url?scp=85054694477&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85054694477&partnerID=8YFLogxK
U2 - 10.1021/acs.analchem.8b03268
DO - 10.1021/acs.analchem.8b03268
M3 - Article
C2 - 30269481
AN - SCOPUS:85054694477
VL - 90
SP - 11756
EP - 11759
JO - Analytical Chemistry
JF - Analytical Chemistry
SN - 0003-2700
IS - 20
ER -