Single Cell Analysis of Proteoforms

Pei Su; Michael A.R. Hollas; Fatma Ayaloglu Butun; Vijaya Lakshmi Kanchustambham; Stanislav Rubakhin; Namrata Ramani; Joseph B. Greer; Bryan P. Early; Ryan T. Fellers; Michael A. Caldwell; Jonathan V. Sweedler; Jared O. Kafader; Neil L. Kelleher

doi:10.1021/acs.jproteome.4c00075

Single Cell Analysis of Proteoforms

Pei Su, Michael A.R. Hollas, Fatma Ayaloglu Butun, Vijaya Lakshmi Kanchustambham, Stanislav Rubakhin, Namrata Ramani, Joseph B. Greer, Bryan P. Early, Ryan T. Fellers, Michael A. Caldwell, Jonathan V. Sweedler, Jared O. Kafader, Neil L. Kelleher^*

^*Corresponding author for this work

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

8 Scopus citations

Abstract

We introduce single cell Proteoform imaging Mass Spectrometry (scPiMS), which realizes the benefit of direct solvent extraction and MS detection of intact proteins from single cells dropcast onto glass slides. Sampling and detection of whole proteoforms by individual ion mass spectrometry enable a scalable approach to single cell proteomics. This new scPiMS platform addresses the throughput bottleneck in single cell proteomics and boosts the cell processing rate by several fold while accessing protein composition with higher coverage.

Original language	English (US)
Pages (from-to)	1883-1893
Number of pages	11
Journal	Journal of Proteome Research
Volume	23
Issue number	6
DOIs	https://doi.org/10.1021/acs.jproteome.4c00075
State	Published - Jun 7 2024

Funding

The authors also thank Prof. Chad Mirkin (Northwestern University) for valuable discussions. High-contrast bright field imaging experiments were performed with the help from Biological Imaging Facility at Northwestern University (RRID:SCR_017767), graciously supported by the Chemistry for Life Processes Institute, the NU Office for Research, the Department of Molecular Biosciences and the Rice Foundation. Funding from National Institutes of Health P41 GM108569 (N.L.K.), National Institutes of Health UH3 CA246635 (N.L.K.), National Institutes of Health P30 DA018310 (N.L.K. and J.V.S.), and National Institutes of Health P30 CA060553 (awarded to the Robert H. Lurie Comprehensive Cancer Center).

Keywords

individual ion mass spectrometry
mass spectrometry
nano-DESI
proteoform assignment score
single cell
top-down proteomics

ASJC Scopus subject areas

General Chemistry
Biochemistry

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10.1021/acs.jproteome.4c00075

Cite this

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title = "Single Cell Analysis of Proteoforms",

abstract = "We introduce single cell Proteoform imaging Mass Spectrometry (scPiMS), which realizes the benefit of direct solvent extraction and MS detection of intact proteins from single cells dropcast onto glass slides. Sampling and detection of whole proteoforms by individual ion mass spectrometry enable a scalable approach to single cell proteomics. This new scPiMS platform addresses the throughput bottleneck in single cell proteomics and boosts the cell processing rate by several fold while accessing protein composition with higher coverage.",

keywords = "individual ion mass spectrometry, mass spectrometry, nano-DESI, proteoform assignment score, single cell, top-down proteomics",

author = "Pei Su and Hollas, {Michael A.R.} and Butun, {Fatma Ayaloglu} and Kanchustambham, {Vijaya Lakshmi} and Stanislav Rubakhin and Namrata Ramani and Greer, {Joseph B.} and Early, {Bryan P.} and Fellers, {Ryan T.} and Caldwell, {Michael A.} and Sweedler, {Jonathan V.} and Kafader, {Jared O.} and Kelleher, {Neil L.}",

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doi = "10.1021/acs.jproteome.4c00075",

language = "English (US)",

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AU - Hollas, Michael A.R.

AU - Butun, Fatma Ayaloglu

AU - Kanchustambham, Vijaya Lakshmi

AU - Rubakhin, Stanislav

AU - Ramani, Namrata

AU - Greer, Joseph B.

AU - Early, Bryan P.

AU - Fellers, Ryan T.

AU - Caldwell, Michael A.

AU - Sweedler, Jonathan V.

AU - Kafader, Jared O.

AU - Kelleher, Neil L.

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AB - We introduce single cell Proteoform imaging Mass Spectrometry (scPiMS), which realizes the benefit of direct solvent extraction and MS detection of intact proteins from single cells dropcast onto glass slides. Sampling and detection of whole proteoforms by individual ion mass spectrometry enable a scalable approach to single cell proteomics. This new scPiMS platform addresses the throughput bottleneck in single cell proteomics and boosts the cell processing rate by several fold while accessing protein composition with higher coverage.

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KW - mass spectrometry

KW - nano-DESI

KW - proteoform assignment score

KW - single cell

KW - top-down proteomics

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Single Cell Analysis of Proteoforms

Abstract

Funding

Keywords

ASJC Scopus subject areas

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