A5088493916- Advanced Proteomics Techniques and Applications
- Mass Spectrometry Techniques and Applications
- Biosensors and Analytical Detection
- Analytical Chemistry and Chromatography
- Microfluidic and Capillary Electrophoresis Applications
- Spectroscopy and Quantum Chemical Studies
- MicroRNA in disease regulation
- Protein Structure and Dynamics
- Metabolomics and Mass Spectrometry Studies
- Lipid Membrane Structure and Behavior
- Epigenetics and DNA Methylation
- Advanced biosensing and bioanalysis techniques
- Ferroptosis and cancer prognosis
- RNA Interference and Gene Delivery
- Traditional Chinese Medicine Analysis
- Reproductive Biology and Fertility
- Advancements in Transdermal Drug Delivery
- Natural product bioactivities and synthesis
- Circular RNAs in diseases
- Cancer Immunotherapy and Biomarkers
- interferon and immune responses
- Monoclonal and Polyclonal Antibodies Research
- Advanced Biosensing Techniques and Applications
- Zebrafish Biomedical Research Applications
- Cytokine Signaling Pathways and Interactions
Guangdong Pharmaceutical University
2023-2025
Michigan State University
2017-2024
People's Government of Guangdong Province
2011-2012
Capillary zone electrophoresis (CZE)-tandem mass spectrometry (MS/MS) has been recognized as a useful tool for top-down proteomics. However, its performance deep proteomics is still dramatically lower than widely used reversed-phase liquid chromatography (RPLC)-MS/MS. We present an orthogonal multidimensional separation platform that couples size exclusion (SEC) and RPLC based protein prefractionation to CZE-MS/MS of Escherichia coli. The generated high peak capacity (∼4000) intact proteins,...
Native proteomics aims to characterize complex proteomes under native conditions and ultimately produces a full picture of endogenous protein complexes in cells. It requires novel analytical platforms for high-resolution liquid-phase separation prior mass spectrometry (MS) MS/MS. In this work, size-exclusion chromatography (SEC)-capillary zone electrophoresis (CZE)-MS/MS was developed discovery mode, resulting the identification 144 proteins, 672 proteoforms, 23 from Escherichia coli...
Better peptide separation is required for bottom-up proteomics further improving the proteome coverage. The two-dimensional liquid chromatography (2D-LC) systems only explore differences among peptides in their hydrophobicity (reversed-phase, RP) and charge (strong cation/anion exchange, SCX/SAX). Alternative techniques with different mechanisms are to improve separation. Capillary zone electrophoresis (CZE) an attractive alternative because it has high efficiency of biomolecules separates...
Native capillary zone electrophoresis-mass spectrometry (CZE-MS) has attracted attentions for the characterization of monoclonal antibodies (mAbs) due to potential CZE highly efficient separations mAbs under native conditions as well its compatibility with electrospray ionization (ESI)-MS. However, low sample loading capacity and limited separation resolution large proteins protein complexes (e.g. mAbs) impede widespread adoption CZE-MS. Here, we present a novel isoelectric focusing...
Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has been well recognized for bottom–up proteomics. It approached 4000–8000 protein identifications (IDs) from a human cell line, mouse brains, or Xenopus embryos via coupling with liquid chromatography (LC) prefractionation. However, at least 500 μg of complex proteome digests were required the LC/CZE-MS/MS studies. This requirement large amount initial peptide material impedes application CZE-MS/MS deep proteomics...
Top-down proteomics (TDP) aims to delineate proteomes in a proteoform-specific manner, which is vital for accurately understanding protein function cellular processes. It requires high-capacity separation of proteoforms before mass spectrometry (MS) and tandem MS (MS/MS). Capillary isoelectric focusing (cIEF)-MS has been recognized as useful tool TDP the 1990s because cIEF capable high-resolution proteoforms. Previous cIEF-MS studies concentrated on measuring protein's without MS/MS,...
Phosphoproteomics requires better separation of phosphopeptides to boost the coverage phosphoproteome. We argue that an alternative method produces orthogonal phosphopeptide widely used LC needs be considered. Capillary zone electrophoresis (CZE) is one important because CZE and are for migration time peptides in can accurately predicted. In this work, we coupled strong cation exchange (SCX)-reversed-phase (RPLC) CZE-MS/MS large-scale phosphoproteomics colon carcinoma HCT116 cell line. The...
Large-scale top-down proteomics characterizes proteoforms in cells globally with high confidence and throughput using reversed-phase liquid chromatography (RPLC)-tandem mass spectrometry (MS/MS) or capillary zone electrophoresis (CZE)-MS/MS. The false discovery rate (FDR) from the target-decoy database search is typically deployed to filter identified ensure high-confidence identifications (IDs). It has been demonstrated that FDRs can be drastically underestimated. An alternative approach...
A universal and standardized sample preparation method becomes vital for denaturing top-down proteomics (dTDP) to advance the scale accuracy of proteoform delineation in complex biological systems. It needs have high protein recovery, minimum bias, good reproducibility, compatibility with downstream mass spectrometry (MS) analysis. Here, we employed a lysis buffer containing sodium dodecyl sulfate extracting proteoforms from cells and, first time, compared membrane ultrafiltration (MU),...
Top-down proteomics (TDP) is an ideal approach for deciphering the histone code and it routinely employs reversed-phase liquid chromatography (RPLC)-tandem mass spectrometry (MS/MS). Because of extreme complexity histones regarding number proteoforms, new analytical tools with high-capacity separation highly sensitive detection proteoforms are required TDP histones. Here we present capillary zone electrophoresis (CZE)-MS/MS via electro-kinetically pumped sheath-flow CE-MS interface...
Production of site-specific cysteine-engineered antibody-drug conjugates (ADCs) in mammalian cells may produce developability challenges, fragments, and heterogenous molecules, leading to potential product critical quality attributes later development stages. Liquid phase chromatography with mass spectrometry (LC-MS) is widely used evaluate antibody impurities drug-to-antibody ratio, but faces challenges analysis fragment variants ADCs oligonucleotide-to-antibody ratio (OAR) species...
Novel mass spectrometry (MS)-based proteomic tools with extremely high sensitivity and peak capacity are required for comprehensive characterization of protein molecules in mass-limited samples. We reported a nanoRPLC-CZE-MS/MS system deep bottom-up proteomics low micrograms human cell samples previous work. In this work, we improved the drastically via employing bovine serum albumin (BSA)-treated sample vials, improving nanoRPLC fraction collection procedure, using short capillary fast CZE...
MicroRNA (miRNA) in urine has been considered as a potential biomarker for early stage diagnosis of multiple diseases like urinary system cancer, kidney injury, and diabetes because their many demonstrated advantages including long-term stability noninvasiveness. However, the traditional enrichment extraction processes miRNAs from are cumbersome tedious due to low concentration carriers miRNAs. Herein, we present novel method collect concentrations dilute solutions such cell culture medium....
Capillary zone electrophoresis-tandem mass spectrometry (CZE-MS/MS) has attracted attention recently for large-scale top-down proteomics that aims to characterize proteoforms in cells at a global scale and with high throughput. In this work, CZE-MS/MS ultraviolet photodissociation (UVPD) was evaluated the first time. Roughly, 600 369 proteins were identified from zebrafish brain sample via coupling size exclusion chromatography (SEC) fractionation CZE-UVPD. The dataset represents one of...
Defining the denatured state ensemble (DSE) and disordered proteins is essential to understanding folding, chaperone action, degradation, translocation. As compared with water-soluble proteins, DSE of membrane much less characterized. Here, we measure helical protein GlpG
Large-scale bottom-up proteomics of few even single cells is crucial for a better understanding the roles played by cell-to-cell heterogeneity in disease and development. Novel proteomic methodologies with extremely high sensitivity are required single-cell proteomics. Sample processing recovery no contaminants one key step. Here we developed nanoparticle-aided nanoreactor nanoproteomics (Nano3) technique low-nanograms mammalian cell proteins proteome profiling. The Nano3 employed...