PXD040455 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | HyperSCP: Combining Isotopic and Isobaric Labeling for Higher Throughput Single-Cell Proteomics |
Description | Recent developments in mass spectrometry-based single-cell proteomics (SCP) have resulted in dramatically improved sensitivity, yet the relatively low measurement throughput remains a limitation. Isobaric and isotopic labeling methods have been separately applied to SCP to increase throughput through multiplexing. Here we combined both forms of labeling to achieve multiplicative scaling for higher throughput. Two-plex stable isotope labeling of amino acids in cell culture and isobaric Tandem Mass Tag labeling enabled up to 32 single cells to be analyzed in a single LC-MS analysis, in addition to reference and negative control channels. A custom nested nanowell chip was used for nanoliter sample processing to minimize sample losses. Using a 145-min total LC-MS cycle time, ~280 single cells were analyzed per day, which could be increased to ~700 samples per day with a high-duty-cycle multicolumn LC system producing the same active gradient. The labeling efficiency and achievable proteome coverage were characterized for multiple analysis conditions. Despite some decrease in coverage, this method readily differentiated four different cell types and thus proves suitable for, e.g., rapid cell typing. |
HostingRepository | PRIDE |
AnnounceDate | 2023-11-14 |
AnnouncementXML | Submission_2023-11-14_09:02:04.466.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Ryan Kelly |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | iodoacetamide derivatized residue |
Instrument | Orbitrap Exploris 480 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-02-27 06:15:09 | ID requested | |
1 | 2023-07-11 12:39:16 | announced | |
⏵ 2 | 2023-11-14 09:02:08 | announced | 2023-11-14: Updated project metadata. |
Publication List
Liang Y, Truong T, Saxton AJ, Boekweg H, Payne SH, Van Ry PM, Kelly RT, HyperSCP: Combining Isotopic and Isobaric Labeling for Higher Throughput Single-Cell Proteomics. Anal Chem, 95(20):8020-8027(2023) [pubmed] |
Keyword List
submitter keyword: multiplex, single-cell proteomics,high-throughput |
Contact List
Ryan T. Kelly |
contact affiliation | Department of Chemistry and Biochemistry, Brigham Young University |
contact email | ryan@chem.byu.edu |
lab head | |
Ryan Kelly |
contact affiliation | Brigham Young University |
contact email | ryan.kelly@byu.edu |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD040455
- Label: PRIDE project
- Name: HyperSCP: Combining Isotopic and Isobaric Labeling for Higher Throughput Single-Cell Proteomics