Evidence map›Paper›PMID 39266194›Full record

ArticleAnalytica chimica acta2024

A simple and efficient approach for preparing cationic coating with tunable electroosmotic flow for capillary zone electrophoresis-mass spectrometry-based top-down proteomics.

Qianjie Wang, Guangyao Gao, Fei Fang, Qianyi Wang, Peter K Lundquist, Liangliang Sun

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In one paragraph

Article in Analytica chimica acta, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.

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6citing papers in PubMed
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1 · What the graph read from it

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2 · The registry

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3 · Its place in the literature

Who cites it

6 citing papers in PubMed.

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4 · The record

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5 · Who and what money

Authors and funding

6 authors.

Qianjie WangDepartment of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA; Department of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA; Plant Resilience Institute, Michigan State University, East Lansing, MI, 48824, USA.
Guangyao GaoDepartment of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA.
Fei FangDepartment of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA.
Qianyi WangDepartment of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA.
Peter K LundquistDepartment of Biochemistry and Molecular Biology, Michigan State University, East Lansing, MI, 48824, USA; Plant Resilience Institute, Michigan State University, East Lansing, MI, 48824, USA.
Liangliang SunDepartment of Chemistry, Michigan State University, 578 S Shaw Lane, East Lansing, MI, 48824, USA. Electronic address: lsun@chemistry.msu.edu.

Funding

Quantitative top-down proteomics of human colorectal cancer cells and tumorsR01CA247863 · NCI · MICHIGAN STATE UNIVERSITY · PI HUMMON, AMANDA B., LIU, XIAOWEN · 2021 to 2025
$1.9M
Advancing top-down proteomics with capillary electrophoresis-mass spectrometryR35GM153479 · NIGMS · MICHIGAN STATE UNIVERSITY · PI Liangliang Sun · 2024 to 2026
$1.4M
NCI NIH HHS R01 CA247863NIGMS NIH HHS R35 GM153479
6 · The paper itself

Abstract

backgroundCapillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has become a valuable analytical technique in top-down proteomics (TDP). CZE-MS/MS-based TDP typically employs separation capillaries with neutral coatings (i.e., linear polyacrylamide, LPA). However, issues related to separation resolution and reproducibility remain with the LPA-coated capillaries due to the unavoidable non-specific protein adsorption onto the capillary wall. Cationic coatings can be critical alternatives to LPA coating for CZE-MS/MS-based TDP due to the electrostatic repulsion between the positively charged capillary inner wall and proteoform molecules in the acidic separation buffer. Unfortunately, there are only very few studies using cationic coating-based CZE-MS/MS for TDP studies.

resultsIn this work, we aimed to develop a simple and efficient approach for preparing separation capillaries with a cationic coating, i.e., poly (acrylamide-co-(3-acrylamidopropyl) trimethylammonium chloride [PAMAPTAC]) for CZE-MS/MS-based TDP. The PAMAPTAC coating-based CZE-MS produced significantly better separation resolution of proteoforms compared to the traditionally used LPA-coated approach. It achieved reproducible separation and measurement of a simple proteoform mixture and a complex proteome sample (i.e., a yeast cell lysate) regarding migration time, proteoform intensity, and the number of proteoform identifications. The PAMAPTAC coating-based CZE-MS enabled the detection of large proteoforms (≥30 kDa) from the yeast cell lysate reproducibly without any size-based prefractionation. Interestingly, the mobility of proteoforms using the PAMAPTAC coating can be predicted accurately using a simple semi-empirical model. SIGNIFICANCE: The results render the PAMAPTAC coating as a valuable alternative to the LPA coating to advance CZE-MS-based TDP towards high-resolution separation and highly reproducible measurement of proteoforms in complex samples.

Indexed as

CationsElectroosmosisElectrophoresis, CapillaryProteomicsSaccharomyces cerevisiaeTandem Mass SpectrometryCationsCapillary zone electrophoresis-mass spectrometryCovalent cationic coatingLarge proteoformsProteoformReproducibilityTop-down proteomics

Identifiers

PMID39266194
PMCPMC11404064

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.