ArticleAnalytical chemistry2023
A High-Sensitivity Low-Nanoflow LC-MS Configuration for High-Throughput Sample-Limited Proteomics.
Article in Analytical chemistry, 2023. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 27 papers.
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Who cites it
27 citing papers in PubMed.
- Optimization of trap column properties and loading conditions for proteome profiling of single-cell-level sample inputs.Analytical and bioanalytical chemistry · 2026Article
- Miniaturized chromatography-based lipidomics methods towards single-cell analysis.Analytical and bioanalytical chemistry · 2026Review
- DDA/DIA mass spectrometry coupled with molecular networking and machine learning: high-coverage profiling of pyrrolizidine alkaloids.Analytical and bioanalytical chemistry · 2026Article
- MALDI-TOF Mass Spectrometry for Glioblastoma Secretome Biomarker Screening: A Review of Challenges and Perspectives.Current issues in molecular biology · 2026Review
- Accurate quantification in proteomics with QuantUMS.Nature biotechnology · 2026Article
- Neu5Gc-associated Serum Immunoglobulin G Glycosylation as a Diagnostic Biomarker for Hepatocellular Carcinoma.Journal of clinical and translational hepatology · 2026Article
- Low-resolution FAIMS for increased peptide coverage in low-load and single-cell proteomics.Scientific reports · 2026Article
- Modification of a Low-Cost Pipetting Robot for Nanoliter Liquid Handling and Autosampling for Liquid Chromatography-Mass Spectrometry.Journal of separation science · 2026Article
- Editorial: Multiomics advances: bridging gaps in understanding intricate biological systems.Frontiers in plant science · 2026Article
- In-depth analysis of data characteristics and comparative evaluation of dda and dia accuracy in label-free quantitative proteomics of biological samples.Clinical proteomics · 2025Article
- High-Throughput Label-free Single-Cell Proteomics Enabled by Multicolumn NanoLC with a 5-min Cycle Time.Research square · 2025Article
- Cost-effective method for semi-quantitative analysis of soluble endoglin in biological samples after anti-endoglin monoclonal antibody treatment.Scientific reports · 2025Article
- Deciphering Radiotherapy Resistance: A Proteomic Perspective.Proteomes · 2025Review
- Data-Independent Acquisition Shortens the Analytical Window of Single-Cell Proteomics to Fifteen Minutes in Capillary Electrophoresis Mass Spectrometry.Journal of proteome research · 2025Article
- Trends in Mass Spectrometry-Based Single-Cell Proteomics.Analytical chemistry · 2025Review
- Challenging the Astral mass analyzer to quantify up to 5,300 proteins per single cell at unseen accuracy to uncover cellular heterogeneity.Nature methods · 2025Article
- Residual host cell proteins: sources, properties, detection methods and data acquisition modes.Frontiers in microbiology · 2025Review
- Simple and robust high-throughput serum proteomics workflow with low-microflow LC-MS/MS.Analytical and bioanalytical chemistry · 2024Article
- Proteomic analysis of plasma exosomes in patients with metastatic colorectal cancer.Clinical proteomics · 2024Article
- Repurposed 3D Printer Allows Economical and Programmable Fraction Collection for Proteomics of Nanogram Scale Samples.Analytical chemistry · 2024Article
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
This work demonstrates the utility of high-throughput nanoLC-MS and label-free quantification (LFQ) for sample-limited bottom-up proteomics analysis, including single-cell proteomics (SCP). Conditions were optimized on a 50 μm internal diameter (I.D.) column operated at 100 nL/min in the direct injection workflow to balance method sensitivity and sample throughput from 24 to 72 samples/day. Multiple data acquisition strategies were also evaluated for proteome coverage, including data-dependent acquisition (DDA), wide-window acquisition (WWA), and wide-window data-independent acquisition (WW-DIA). Analyzing 250 pg HeLa digest with a 10-min LC gradient (72 samples/day) provided >900, >1,800, and >3,000 protein group identifications for DDA, WWA, and WW-DIA, respectively. Total method cycle time was further reduced from 20 to 14.4 min (100 samples/day) by employing a trap-and-elute workflow, enabling 70% mass spectrometer utilization. The method was applied to library-free DIA analysis of single-cell samples, yielding >1,700 protein groups identified. In conclusion, this study provides a high-sensitivity, high-throughput nanoLC-MS configuration for sample-limited proteomics.
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