CoChat
Sign Up
Cited by 112Advanced Proteomics Techniques and ApplicationsMass Spectrometry Techniques and ApplicationsMicrofluidic and Capillary Electrophoresis Applications

Recent trends of capillary electrophoresis‐mass spectrometry in proteomics research

Open access
Gomes, Fabio P., Yates, John R.·2019·Mass Spectrometry Reviews
DOI 10.1002/mas.21599PMC PMC6800771
Open this paper in CoChat — free account

Abstract

Progress in proteomics research has led to a demand for powerful analytical tools with high separation efficiency and sensitivity for confident identification and quantification of proteins, posttranslational modifications, and protein complexes expressed in cells and tissues. This demand has significantly increased interest in capillary electrophoresis‐mass spectrometry (CE‐MS) in the past few years. This review provides highlights of recent advances in CE‐MS for proteomics research, including a short introduction to top‐down mass spectrometry and native mass spectrometry (native MS), as well as a detailed overview of CE methods. Both the potential and limitations of these methods for the analysis of proteins and peptides in synthetic and biological samples and the challenges of CE methods are discussed, along with perspectives about the future direction of CE‐MS. @ 2019 Wiley Periodicals, Inc. Mass Spec Rev 00:1–16, 2019.

Integrity signals

Registry

Crossref · 10.1002/mas.21599

Cites

141 of 141 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

0 of 181 cited sources checked · 0 supported · 0 flagged

  • Premisecitation

    Solvent consumption in CE-MS is very low, making it environmentally advantageous.

    “In addition, solvent consumption (aqueous or organic) in CE-MS is very low, making it environmentally advantageous9.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [9] Tycova A, Ledvina V, Kleparnik K. Recent advances in CE-MS coupling: Instrumentation, methodology, and applications. Electrophoresis. 2017; 38(1): 115– 134. 27783411 source

      Not checked in the free run.

  • Premisecitation

    Confident identification and quantification of proteins, PTMs, and peptides in biological samples depends on rigorous peptide separation before mass spectrometry.

    “Confident identification and quantification of proteins, posttranslational modifications (PTMs), and peptides in biological samples rely on rigorous separation of peptides prior to introduction into the mass spectrometer1,5,6.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [1] Zhang Y, Fonslow BR, Shan B, Baek MC, Yates JR 3rd. Protein analysis by shotgun/bottom-up proteomics. Chemical reviews. 2013; 113(4): 2343– 2394. 23438204 PMC3751594 source

      Not checked in the free run.

    • cites [5] Fonslow BR, Yates JR 3rd. Capillary electrophoresis applied to proteomic analysis. Journal of separation science. 2009; 32(8): 1175– 1188. 19360788 PMC3509210 source

      Not checked in the free run.

    • cites [6] Aebersold R, Mann M. Mass-spectrometric exploration of proteome structure and function. Nature. 2016; 537(7620): 347– 355. 27629641 source

      Not checked in the free run.

  • Premisecitation

    Recent advances in CE-MS platforms have attracted considerable interest from the proteomics community as an alternative to LC-MS.

    “Recent advances in CE-MS platforms have attracted considerable interest from the proteomics community as an alternative to LC-MS9–11.”
    Quote in the paperSubstantial claim0 of 3 sources checked
    • cites [9] Tycova A, Ledvina V, Kleparnik K. Recent advances in CE-MS coupling: Instrumentation, methodology, and applications. Electrophoresis. 2017; 38(1): 115– 134. 27783411 source

      Not checked in the free run.

    • cites [10] Stolz A, Jooss K, Hocker O, Romer J, Schlecht J, Neususs C. Recent advances in capillary electrophoresis-mass spectrometry: instrumentation, methodology and applications. Electrophoresis. 2018. 30260009 source

      Not checked in the free run.

    • cites [11] Bush DR, Zang L, Belov AM, Ivanov AR, Karger BL. High Resolution CZE-MS Quantitative Characterization of Intact Biopharmaceutical Proteins: Proteoforms of Interferon-beta1. Analytical chemistry. 2016; 88(2): 1138– 1146. 26641950 source

      Not checked in the free run.

  • Interpretationreasoning

    Peptide-based methods may limit understanding of the intricate control of diverse biological activities by PTMs.

    “Consequently, peptide-based methods may limit our understanding of the intricate and nuanced control of diverse biological activities by PTMs14.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [14] Toby TK, Fornelli L, Kelleher NL. Progress in Top-Down Proteomics and the Analysis of Proteoforms. Annual review of analytical chemistry (Palo Alto, Calif). 2016; 9(1): 499– 519. PMC5373801 27306313 source

      Not checked in the free run.

  • Premisecitation

    Recent improvements in mass spectrometry instrumentation and bioinformatics have enabled TDMS to characterize and quantify hundreds to thousands of proteoforms in a single analysis.

    “Recent improvements in mass spectrometry instrumentation and bioinformatics have enabled TDMS to be used to characterize and quantify hundreds to thousands of proteoforms in a single analysis16.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Chen B, Brown KA, Lin Z, Ge Y. Top-Down Proteomics: Ready for Prime Time? Analytical chemistry. 2018; 90(1): 110– 127. 29161012 PMC6138622 source

      Not checked in the free run.

  • Classified as probably not a substantial claim. 20
  • May combine two claims. 57

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Mass spectrometry-based methods remain the most widely used approach for high-throughput proteomics research.

    “Mass spectrometry (MS)–based methods continue to be the most widely used methods for high-throughput proteomics research1–4”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [1] Zhang Y, Fonslow BR, Shan B, Baek MC, Yates JR 3rd. Protein analysis by shotgun/bottom-up proteomics. Chemical reviews. 2013; 113(4): 2343– 2394. 23438204 PMC3751594 source

      Not checked in the free run.

    • cites [2] Yates JR, Ruse CI, Nakorchevsky A. Proteomics by mass spectrometry: approaches, advances, and applications. Annual review of biomedical engineering. 2009; 11: 49– 79. 19400705 source

      Not checked in the free run.

    • cites [3] Nilsson T, Mann M, Aebersold R, Yates JR 3rd, Bairoch A, Bergeron JJ. Mass spectrometry in high-throughput proteomics: ready for the big time. Nature methods. 2010; 7(9): 681– 685. 20805795 source

      Not checked in the free run.

    • cites [4] Yates JR 3rd. The revolution and evolution of shotgun proteomics for large-scale proteome analysis. Journal of the American Chemical Society. 2013; 135(5): 1629– 1640. 23294060 PMC3751590 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Capillary electrophoresis is a mature separation technology that couples effectively with mass spectrometry and is well suited for peptide separation.

    “Capillary electrophoresis is a mature separation technology that is effectively coupled with mass spectrometry (CE-MS) and is well suited for this purpose7,8.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [7] Mischak H, Coon JJ, Novak J, Weissinger EM, Schanstra JP, Dominiczak AF. Capillary electrophoresis-mass spectrometry as a powerful tool in biomarker discovery and clinical diagnosis: an update of recent developments. Mass spectrometry reviews. 2009; 28(5): 703– 724. 18973238 PMC2720435 source

      Not checked in the free run.

    • cites [8] Belczacka I, Latosinska A, Siwy J, Urinary CE-MS peptide marker pattern for detection of solid tumors. Scientific reports. 2018; 8(1): 5227– 5227. 29588543 PMC5869723 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Native MS provides a proteomics tool that enables identification and characterization of biomolecular structures and advances understanding of biological pathways at the molecular level.

    “Native MS provides a proteomics tool that enables identification and characterization of these biomolecular structures and advances the understanding of their biological pathways at the molecular level19,20,35.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [19] Li H, Nguyen HH, Ogorzalek Loo RR, Campuzano IDG, Loo JA. An integrated native mass spectrometry and top-down proteomics method that connects sequence to structure and function of macromolecular complexes. Nature chemistry. 2018; 10(2): 139– 148. PMC5784781 29359744 source

      Not checked in the free run.

    • cites [20] Leney AC, Heck AJ. Native Mass Spectrometry: What is in the Name? Journal of the American Society for Mass Spectrometry. 2017; 28(1): 5– 13. 27909974 PMC5174146 source

      Not checked in the free run.

    • cites [35] Belov AM, Viner R, Santos MR, Analysis of Proteins, Protein Complexes, and Organellar Proteomes Using Sheathless Capillary Zone Electrophoresis - Native Mass Spectrometry. Journal of the American Society for Mass Spectrometry. 2017; 28(12): 2614– 2634. 28875426 PMC5709234 source

      Not checked in the free run.

    • Rests on an assumption: Most proteins are assembled into protein complexes to achieve functional activity.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Capillary isoelectric focusing is a useful but less commonly used CE technique in proteomics.

    “Capillary isoelectric focusing (cIEF)12 is another useful, but less commonly used CE technique.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [12] Kahle J, Stein M, Watzig H. Design of experiments as a valuable tool for biopharmaceutical analysis with (imaged) capillary isoelectric focusing. Electrophoresis. 2019. 31177564 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Top-down analysis can provide unique structural insights into proteins through precise molecular weight measurements, identification of multiple PTMs, and in-depth proteoform characterization.

    “Top-down analysis can provide unique insights into a protein’s structure by providing precise molecular weight measurements, the identification of multiple PTMs5,16,28 and in-depth proteoform characterization (different forms of a single protein generated from a gene)32.”
    Quote in the paperSubstantial claim0 of 4 sources checked
    • cites [5] Fonslow BR, Yates JR 3rd. Capillary electrophoresis applied to proteomic analysis. Journal of separation science. 2009; 32(8): 1175– 1188. 19360788 PMC3509210 source

      Not checked in the free run.

    • cites [16] Chen B, Brown KA, Lin Z, Ge Y. Top-Down Proteomics: Ready for Prime Time? Analytical chemistry. 2018; 90(1): 110– 127. 29161012 PMC6138622 source

      Not checked in the free run.

    • cites [28] Haselberg R, de Jong GJ, Somsen GW. CE-MS for the analysis of intact proteins 2010–2012. Electrophoresis. 2013; 34(1): 99– 112. 23161520 source

      Not checked in the free run.

    • cites [32] Smith LM, Kelleher NL. Proteoform: a single term describing protein complexity. Nature methods. 2013; 10(3): 186– 187. 23443629 PMC4114032 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Top-down mass spectrometry is an emerging approach that provides detailed structural information on intact proteins, including PTM characterization.

    “TDMS is an emerging approach that provides detailed structural information on intact proteins, including characterization of PTMs16.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [16] Chen B, Brown KA, Lin Z, Ge Y. Top-Down Proteomics: Ready for Prime Time? Analytical chemistry. 2018; 90(1): 110– 127. 29161012 PMC6138622 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Native MS can interrogate intact proteins in a near native state while preserving noncovalent protein-protein and protein-ligand complexes.

    “Native MS can be used to interrogate the structure of intact proteins in their near native state, with an emphasis on preserving noncovalent protein-protein and protein-ligand complexes20.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [20] Leney AC, Heck AJ. Native Mass Spectrometry: What is in the Name? Journal of the American Society for Mass Spectrometry. 2017; 28(1): 5– 13. 27909974 PMC5174146 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Bottom-up approaches are high-throughput, but critical information about PTMs and subunit protein assemblies is generally lost after proteolysis.

    “This approach is well-known for its high-throughput performance; however, critical information about PTMs and subunit protein assemblies (via non-covalent interactions) is generally lost after proteolysis14,31.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [14] Toby TK, Fornelli L, Kelleher NL. Progress in Top-Down Proteomics and the Analysis of Proteoforms. Annual review of analytical chemistry (Palo Alto, Calif). 2016; 9(1): 499– 519. PMC5373801 27306313 source

      Not checked in the free run.

    • cites [31] Catherman AD, Skinner OS, Kelleher NL. Top Down proteomics: facts and perspectives. Biochemical and biophysical research communications. 2014; 445(4): 683– 693. 24556311 PMC4103433 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Native MS provides information about ligand binding, dynamics, subunit stoichiometry, stability, topology, and assembly structural properties.

    “Native MS provides information about ligand binding, dynamics, stoichiometry of individual subunits, stability, topology, and assembly structural properties35.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [35] Belov AM, Viner R, Santos MR, Analysis of Proteins, Protein Complexes, and Organellar Proteomes Using Sheathless Capillary Zone Electrophoresis - Native Mass Spectrometry. Journal of the American Society for Mass Spectrometry. 2017; 28(12): 2614– 2634. 28875426 PMC5709234 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    PTM patterns can provide insight into potentially significant biological processes.

    “PTM patterns can provide insight into potentially significant biological processes14,16,17”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [14] Toby TK, Fornelli L, Kelleher NL. Progress in Top-Down Proteomics and the Analysis of Proteoforms. Annual review of analytical chemistry (Palo Alto, Calif). 2016; 9(1): 499– 519. PMC5373801 27306313 source

      Not checked in the free run.

    • cites [16] Chen B, Brown KA, Lin Z, Ge Y. Top-Down Proteomics: Ready for Prime Time? Analytical chemistry. 2018; 90(1): 110– 127. 29161012 PMC6138622 source

      Not checked in the free run.

    • cites [17] Aebersold R, Agar JN, Amster IJ, How many human proteoforms are there? Nature chemical biology. 2018; 14(3): 206– 214. 29443976 PMC5837046 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    The sheathless interface has been widely used for ultrasensitive proteomics analysis, although it is limited by potential pore blocking.

    “The sheathless interface has been widely used in proteomics research to provide ultrasensitive analysis56–58, although it is limited by potential blocking of the pores.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [56] Kammeijer GS, Kohler I, Jansen BC, Dopant Enriched Nitrogen Gas Combined with Sheathless Capillary Electrophoresis-Electrospray Ionization-Mass Spectrometry for Improved Sensitivity and Repeatability in Glycopeptide Analysis. Analytical chemistry. 2016; 88(11): 5849– 5856. 27119460 source

      Not checked in the free run.

    • cites [57] Snyder CM, Zhou X, Karty JA, Fonslow BR, Novotny MV, Jacobson SC. Capillary electrophoresis-mass spectrometry for direct structural identification of serum N-glycans. Journal of chromatography A. 2017; 1523: 127– 139. 28989033 PMC5691604 source

      Not checked in the free run.

    • cites [58] Medina-Casanellas S, Domínguez-Vega E, Benavente F, Sanz-Nebot V, Somsen GW, de Jong GJ. Low-picomolar analysis of peptides by on-line coupling of fritless solid-phase extraction to sheathless capillary electrophoresis-mass spectrometry. Journal of Chromatography A. 2014; 1328: 1– 6. 24438833 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    The Kelleher group reported a standard operating procedure for native MS.

    “Recently, the Kelleher group reported a standard operating procedure for native MS38.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [38] Schachner LF, Ives AN, McGee JP, Standard Proteoforms and Their Complexes for Native Mass Spectrometry. Journal of the American Society for Mass Spectrometry. 2019. PMC6592724 30963455 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Shen and co-workers demonstrated SEC-CZE for analysis of protein complexes and proteoforms under near native conditions, identifying 144 proteins, 672 proteoforms, and 23 protein complexes from the Escherichia coli proteome.

    “More recently, Shen and co-workers demonstrated the use of SEC-CZE for analysis of protein complexes and proteoforms under near native conditions21, resulting in the identification of 144 proteins, 672 proteoforms, and 23 protein complexes from the Escherichia coli proteome.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [21] Shen X, Kou Q, Guo R, Native Proteomics in Discovery Mode Using Size-Exclusion Chromatography-Capillary Zone Electrophoresis-Tandem Mass Spectrometry. Analytical chemistry. 2018; 90(17): 10095– 10099. 30085653 PMC6156775 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Qu and co-workers reported an LC-CZE strategy for site-specific glycan heterogeneity in which CZE analysis of reversed-phase LC glycopeptide fractions reduced sample complexity and enabled glycan structural elucidation.

    “Qu and co-workers reported a LC-CZE strategy for the characterization of site-specific glycan heterogeneity where the use of CZE for analysis of reversed-phase LC glycopeptide fractions reduced the complexity of the sample and enabled glycan structural elucidation88.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [88] Qu Y, Sun L, Zhang Z, Dovichi NJ. Site-Specific Glycan Heterogeneity Characterization by Hydrophilic Interaction Liquid Chromatography Solid-Phase Extraction, Reversed-Phase Liquid Chromatography Fractionation, and Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry. Analytical chemistry. 2018; 90(2): 1223– 1233. 29231704 PMC5771954 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    High peak capacity and resolving power improved data acquisition and enhanced representation of proteins in the sample.

    “High peak capacity and resolving power improved the data acquisition, which resulted in an enhanced representation of the proteins in the sample.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [51] McCool EN, Lubeckyj RA, Shen X, Deep Top-Down Proteomics Using Capillary Zone Electrophoresis-Tandem Mass Spectrometry: Identification of 5700 Proteoforms from the Escherichia coli Proteome. Analytical chemistry. 2018; 90(9): 5529– 5533. 29620868 PMC5930140 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Notable CZE-MS examples include intact mass analysis of biopharmaceutical proteins and monoclonal antibodies, where complete baseline separation is often achieved.

    “Notable examples of CZE-MS include the intact mass analysis of biopharmaceutical proteins11 and monoclonal antibodies22,68 where complete baseline separation is often achieved.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [11] Bush DR, Zang L, Belov AM, Ivanov AR, Karger BL. High Resolution CZE-MS Quantitative Characterization of Intact Biopharmaceutical Proteins: Proteoforms of Interferon-beta1. Analytical chemistry. 2016; 88(2): 1138– 1146. 26641950 source

      Not checked in the free run.

    • cites [22] Han M, Rock BM, Pearson JT, Rock DA. Intact mass analysis of monoclonal antibodies by capillary electrophoresis-Mass spectrometry. Journal of chromatography B, Analytical technologies in the biomedical and life sciences. 2016; 1011: 24– 32. 26751590 source

      Not checked in the free run.

    • cites [68] Zhao Y, Sun L, Knierman MD, Dovichi NJ. Fast separation and analysis of reduced monoclonal antibodies with capillary zone electrophoresis coupled to mass spectrometry. Talanta. 2016; 148: 529– 533. 26653481 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Multi-phosphorylated peptides were poorly retained on the reversed-phase LC-MS stationary phase, leading to fewer peptide identifications than in CE-MS analysis.

    “These authors observed that multi-phosphorylated peptides were poorly retained on the stationary phase of a reversed-phase LC-MS method, resulting in a decreased number of peptide identification relative to the CE-MS analysis.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [84] Faserl K, Sarg B, Gruber P, Lindner HH. Investigating capillary electrophoresis-mass spectrometry for the analysis of common post-translational modifications. Electrophoresis. 2018; 39(9–10): 1208– 1215. 29389038 PMC6001557 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    In the Zhang study, limitations included marginal identifications during the first 20 minutes and a dramatic reduction in identifications after 85 minutes.

    “Despite the high number of identifications, some limitations were observed, including marginal identifications during the first 20 min of the separation, and a dramatic reduction in identifications after 85 min.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [46] Zhang Z, Hebert AS, Westphall MS, Qu Y, Coon JJ, Dovichi NJ. Production of Over 27 000 Peptide and Nearly 4400 Protein Identifications by Single-Shot Capillary-Zone Electrophoresis-Mass Spectrometry via Combination of a Very-Low-Electroosmosis Coated Capillary, a Third-Generation Electrokinetically-Pumped Sheath-Flow Nanospray Interface, an Orbitrap Fusion Lumos Tribrid Mass Spectrometer, and an Advanced-Peak-Determination Algorithm. Analytical chemistry. 2018; 90(20): 12090– 12093. 30179504 PMC6508074 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Li and co-workers reported a bottom-up strategy combining CZE-MS and reversed-phase LC-MS data, which improved the number of identified unique peptides, with hydrophilic and basic low-molecular-weight peptides predominantly identified by CZE-MS.

    “For instance, Li and co-workers reported a bottom-up strategy that combines the data from CZE-MS and reversed-phase LC-MS91. This combination resulted in an improved number of identified unique peptides where hydrophilic and basic peptides with low molecular weight were predominantly identified via CZE-MS.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [91] Li Y, Champion MM, Sun L, Champion PAD, Wojcik R, Dovichi NJ. Capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry as an alternative proteomics platform to ultraperformance liquid chromatography-electrospray ionization-tandem mass spectrometry for samples of intermediate complexity. Analytical chemistry. 2012; 84(3): 1617– 1622. 22182061 PMC3277681 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    According to modeling data, phosphorylation reduces a peptide's charge and thereby its electrophoretic mobility.

    “According to this modeling data, phosphorylation reduces the charge of a peptide, thereby its electrophoretic mobility.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [86] Chen D, Ludwig KR, Krokhin OV, Capillary Zone Electrophoresis-Tandem Mass Spectrometry for Large-Scale Phosphoproteomics with the Production of over 11,000 Phosphopeptides from the Colon Carcinoma HCT116 Cell Line. Analytical chemistry. 2019; 91(3): 2201– 2208. 30624053 PMC6506858 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    CZE separation has frequently been used to analyze glycosylated peptides and glycans, and CZE via TDMS has been used for glycosylation characterization.

    “CZE separation has frequently been used to analyze glycosylated peptides and glycans49,57,80,81 and CZE separation via TDMS was used for glycosylation characterization82.”
    Quote in the paperNot a substantial claim0 of 5 sources checked
    • cites [49] Qu Y, Sun L, Zhu G, Zhang Z, Peuchen EH, Dovichi NJ. Sensitive and fast characterization of site-specific protein glycosylation with capillary electrophoresis coupled to mass spectrometry. Talanta. 2018; 179: 22– 27. 29310225 PMC5763510 source

      Not checked in the free run.

    • cites [57] Snyder CM, Zhou X, Karty JA, Fonslow BR, Novotny MV, Jacobson SC. Capillary electrophoresis-mass spectrometry for direct structural identification of serum N-glycans. Journal of chromatography A. 2017; 1523: 127– 139. 28989033 PMC5691604 source

      Not checked in the free run.

    • cites [80] Giorgetti J, D’Atri V, Canonge J, Monoclonal antibody N-glycosylation profiling using capillary electrophoresis - Mass spectrometry: Assessment and method validation. Talanta. 2018; 178: 530– 537. 29136858 source

      Not checked in the free run.

    • cites [81] Konermann L Addressing a Common Misconception: Ammonium Acetate as Neutral pH “Buffer” for Native Electrospray Mass Spectrometry. Journal of the American Society for Mass Spectrometry. 2017; 28(9): 1827– 1835. 28710594 source

      Not checked in the free run.

    • cites [82] Yang Y, Franc V, Heck AJR. Glycoproteomics: A Balance between High-Throughput and In-Depth Analysis. Trends in Biotechnology. 2017; 35(7): 598– 609. 28527536 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Post-translational phosphorylation plays critical roles in biological systems and can influence the electrophoretic mobility of peptides.

    “Post-translational phosphorylation plays critical roles in biological systems83 and it can also influence the electrophoretic mobility of peptides.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [83] Mou S, Sun L, Dovichi NJ. Accurate determination of peptide phosphorylation stoichiometry via automated diagonal capillary electrophoresis coupled with mass spectrometry: proof of principle. Analytical chemistry. 2013; 85(22): 10692– 10696. 24144020 PMC3873722 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    CZE has proven valuable for protein identification via bottom-up proteomics, where ultrasensitive and fast analysis can be obtained.

    “CZE has also proven to be a valuable tool for protein identification via bottom-up proteomics where ultrasensitive and fast analysis can be obtained72–74.”
    Quote in the paperNot a substantial claim0 of 3 sources checked
    • cites [72] Sun L, Zhu G, Zhao Y, Yan X, Mou S, Dovichi NJ. Ultrasensitive and fast bottom-up analysis of femtogram amounts of complex proteome digests. Angewandte Chemie (International ed in English). 2013; 52(51): 13661– 13664. 24173663 PMC3904452 source

      Not checked in the free run.

    • cites [73] Zhang Z, Dovichi NJ. Optimization of mass spectrometric parameters improve the identification performance of capillary zone electrophoresis for single-shot bottom-up proteomics analysis. Analytica chimica acta. 2018; 1001: 93– 99. 29291811 PMC6159893 source

      Not checked in the free run.

    • cites [74] Zhu G, Sun L, Heidbrink-Thompson J, Capillary zone electrophoresis tandem mass spectrometry detects low concentration host cell impurities in monoclonal antibodies. Electrophoresis. 2016; 37(4): 616– 622. 26530276 PMC4816075 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Ludwig and co-workers reported a CZE-MS approach that identified over 2,300 phosphorylated peptides in a single analysis, illustrating advantages of CZE over LC for PTM analysis.

    “Ludwig and co-workers reported a CZE-MS approach that was able to identify over 2,300 phosphorylated peptides in a single analysis85, thus illustrating the advantages of CZE separation over LC for PTM analysis.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [85] Ludwig KR, Sun L, Zhu G, Dovichi NJ, Hummon AB. Over 2300 phosphorylated peptide identifications with single-shot capillary zone electrophoresis-tandem mass spectrometry in a 100 min separation. Analytical chemistry. 2015; 87(19): 9532– 9537. 26399161 PMC4605816 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationcitation

    The Yang et al. results demonstrated increased sensitivity over C18 ZipTip-only analysis and highlighted advantages of an LC-CZE-MS bottom-up approach.

    “These results demonstrated an increase in sensitivity over analysis after separation using C18 ZipTip only, and highlighted the advantages of using a LC-CZE-MS bottom-up approach.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [47] Yang Z, Shen X, Chen D, Sun L. Microscale Reversed-Phase Liquid Chromatography/Capillary Zone Electrophoresis-Tandem Mass Spectrometry for Deep and Highly Sensitive Bottom-Up Proteomics: Identification of 7500 Proteins with Five Micrograms of an MCF7 Proteome Digest. Analytical chemistry. 2018; 90(17): 10479– 10486. 30102516 PMC6156779 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Glycosylation is an important and common PTM, but its detailed characterization is challenging because of heterogeneity in localization and forms.

    “Glycosylation is an important and common PTM in biological systems, but its detailed characterization is challenging because of its heterogeneity in localization and forms76–79.”
    Quote in the paperNot a substantial claim0 of 4 sources checked
    • cites [76] Mechref Y, Novotny MV. Glycomic analysis by capillary electrophoresis-mass spectrometry. Mass spectrometry reviews. 2009; 28(2): 207– 222. 18973241 source

      Not checked in the free run.

    • cites [77] Jayo RG, Thaysen-Andersen M, Lindenburg PW, Simple capillary electrophoresis-mass spectrometry method for complex glycan analysis using a flow-through microvial interface. Analytical chemistry. 2014; 86(13): 6479– 6486. 24873509 source

      Not checked in the free run.

    • cites [78] Lu G, Crihfield CL, Gattu S, Veltri LM, Holland LA. Capillary Electrophoresis Separations of Glycans. Chemical reviews. 2018; 118(17): 7867– 7885. 29528644 PMC6135675 source

      Not checked in the free run.

    • cites [79] Mechref Y Analysis of glycans derived from glycoconjugates by capillary electrophoresis-mass spectrometry. Electrophoresis. 2011; 32(24): 3467– 3481. 22180203 PMC3360420 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Acetic acid increased protein migration time, which may be attributed to lower pH causing increased hydrodynamic protein radii, more protein unfolding, and reduced electroosmotic flow.

    “The use of acetic acid also increased the migration time of the proteins, which might be attributed to the lower pH of acetic acid that resulted in an increase in hydrodynamics in protein radii, exacerbation of protein unfolding, and reduction of EOF.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [69] Lubeckyj RA, McCool EN, Shen X, Kou Q, Liu X, Sun L. Single-Shot Top-Down Proteomics with Capillary Zone Electrophoresis-Electrospray Ionization-Tandem Mass Spectrometry for Identification of Nearly 600 Escherichia coli Proteoforms. Analytical chemistry. 2017; 89(22): 12059– 12067. 29064224 PMC5726253 source

      Not checked in the free run.

  • May combine two claims.

    Premisenone given

    High ionic strength in the BGE can increase diffusion of the sample zone and cause high current and temperature in the separation capillary, even though it helps avoid band broadening.

    “Although a BGE with high ionic strength is useful to avoid band broadening, high ionic strength can increase the diffusion of the sample zone and lead to high current and temperature in the separation capillary.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Adding organic solvents such as propan-2-ol to an acidic BGE enhances separation efficiency and MS detection, as demonstrated by the Yates group in top-down analysis of Pyrococcus furiosus proteins with a sheathless interface.

    “Interestingly, addition of organic solvents (e.g., propan-2-ol) to an acidic BGE enhances the separation efficiency and MS detection, as demonstrated by Yates group in a top-down analysis of Pyrococcus furiosus proteins using a sheathless interface59.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [59] Han X, Wang Y, Aslanian A, In-line separation by capillary electrophoresis prior to analysis by top-down mass spectrometry enables sensitive characterization of protein complexes. Journal of proteome research. 2014; 13(12): 6078– 6086. 25382489 PMC4262260 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    A 130 nL pH junction injection of BSA identified 40 peptides and produced 70% coverage.

    “In this study, a pH junction injection of 130 nL of BSA resulted in the identification of 40 peptides and produced 70% coverage.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [119] Zhu G, Sun L, Yan X, Dovichi NJ. Bottom-up proteomics of Escherichia coli using dynamic pH junction preconcentration and capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry. Analytical chemistry. 2014; 86(13): 6331– 6336. 24852005 PMC4082393 source

      Not checked in the free run.

    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    cIEF has also been reported for analysis of other intact proteins.

    “cIEF has been also reported for the analysis of other intact proteins94.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [94] Zhu G, Sun L, Dovichi NJ. Simplified capillary isoelectric focusing with chemical mobilization for intact protein analysis. Journal of separation science. 2017; 40(4): 948– 953. 27935257 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    At pH below about 3.5, migration is predominantly influenced by basic residues.

    “For instance, in a pH lower than ~3.5, migration is predominantly influenced by the basic residues.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisenone given

    At low pH, free silanols in the capillary wall are protonated, and protein and peptide separation is enhanced by reduced electroosmotic flow and reduced electrostatic interactions.

    “At low pH the free silanols in the capillary wall are protonated and the separation of proteins and peptides is enhanced due to a reduction of the electroosmotic flow (EOF) and reduction of electrostatic interactions.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Premisenone given

    cIEF is not compatible with a sheathless interface.

    “Also, it is not compatible with a sheathless interface.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Thirty proteins were identified from P. aeruginosa PA01 whole cell lysate using a PA coated capillary.

    “In this study, 30 proteins were identified from P. aeruginosa PA01 whole cell lysate110.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [110] Li Y, Compton PD, Tran JC, Ntai I, Kelleher NL. Optimizing capillary electrophoresis for top-down proteomics of 30–80 kDa proteins. Proteomics. 2014; 14(10): 1158– 1164. 24596178 PMC4034378 source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning

    The sheath-liquid interface is more flexible, and because acidic pH can be obtained from the sheath-liquid reservoir, the BGE pH range can vary between 1 and 8.

    “Conversely, sheath-liquid interface is more flexible and an acidic pH can be obtained from the sheath-liquid reservoir, so the BGE pH range can vary between 1 and 8 units.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisereasoning

    Positively charged coatings require reversed polarity with negative voltage at the inlet, causing proteins and peptides to migrate opposite to EOF.

    “The use of positively charged coatings requires the application of negative voltage on the inlet (reversed polarity); as a result, proteins and peptides will migrate in the opposite direction of the EOF.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisenone given

    Dynamic pH junction relies on preparing samples in a basic buffer, pressure injection into the capillary, and under an electric field negatively charged proteins or peptides migrating toward the proximal tip and being neutralized by an acidic BGE.

    “This stacking technique relies on the preparation of samples in a basic buffer and injection (with pressure) into the separation capillary. Under application of an electric field, negatively charged proteins or peptides migrate toward the proximal tip of the capillary and are neutralized by an acidic BGE.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premisecitation

    In dynamic pH junction, a focused plug forms and target analytes become positively charged and migrate toward the ESI source.

    “A focused plug is formed and target analytes become positively charged, migrating towards the ESI source25,118.”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites [25] Sun L, Zhu G, Yan X, Capillary zone electrophoresis for bottom‐up analysis of complex proteomes. 2016; 16(2): 188– 196. PMC4771184 26508368 source

      Not checked in the free run.

    • cites [118] Zhu G, Sun L, Dovichi NJ. Dynamic pH junction preconcentration in capillary electrophoresis- electrospray ionization-mass spectrometry for proteomics analysis. The Analyst. 2016; 141(18): 5216– 5220. 27460877 PMC5007160 source

      Not checked in the free run.

    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premisecitation

    In protein ESI-MS with holomyoglobin under native conditions, high charge states formed and their signal intensities increased with buffer ionic strength.

    “As illustrated in Figure 6, high charge states (as in denaturing conditions) were formed in native conditions and their signal intensities increased proportionally to the increase in buffer ionic strength.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [101] Hedges JB, Vahidi S, Yue X, Konermann L. Effects of ammonium bicarbonate on the electrospray mass spectra of proteins: evidence for bubble-induced unfolding. Analytical chemistry. 2013; 85(13): 6469– 6476. 23724896 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    Bare fused silica capillaries gave marginal separation efficiency for intact proteins and protein complexes because of protein adsorption, whereas neutral coated capillaries gave high separation efficiency by preventing adsorption.

    “The bare fused silica capillary produced a marginal separation efficiency for intact proteins and protein complexes as a result of protein adsorption to the capillary wall, whereas the neutral coated capillary generated high separation efficiency by preventing protein adsorption35.”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [35] Belov AM, Viner R, Santos MR, Analysis of Proteins, Protein Complexes, and Organellar Proteomes Using Sheathless Capillary Zone Electrophoresis - Native Mass Spectrometry. Journal of the American Society for Mass Spectrometry. 2017; 28(12): 2614– 2634. 28875426 PMC5709234 source

      Not checked in the free run.

    • Rests on an assumption: Hydrophobic and/or ionic interactions between proteins and capillary wall surfaces can increase protein retention or cause peak tailing.
    • Rests on an assumption: Coated capillaries can limit protein adsorption on the capillary wall.

  • May combine two claims.

    Premisenone given

    Ammonium acetate dissolved in water can reach pH 6.8-7.4, although its buffering capacity is low, with buffer ranges at pH 3.8-5.8 and 8.2-10.20.

    “Ammonium acetate can achieve a pH between 6.8 and 7.4 when dissolved in water and despite a low buffering capacity (ammonium acetate buffers in the pH ranges of 3.8–5.8 and 8.2–10.20, see Table 2), it is often used in native MS.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Interpretationreasoning

    Acetonitrile may account for the fast separation and propan-2-ol may provide better resolution of degradation products.

    “Acetonitrile might account for the fast separation and propan-2-ol may provide better resolution of the degradation products.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisereasoning

    In CE-MS applications, the type of interface should be considered when choosing a BGE.

    “In CE-MS applications, the type of interface should also be taken into consideration while choosing a BGE.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    Adding organic solvent to the BGE reduces electroosmotic flow, which improves resolution.

    “The addition of organic solvent to the BGE reduces the EOF, which results in improved resolution.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisereasoning

    BGE selection is most critical for a sheathless interface because the BGE is the only liquid during ESI and acidic pH is recommended to facilitate protein or peptide ionization.

    “BGE selection is most critical for a sheathless interface because the BGE is the only liquid present during the ESI process and acidic pH is recommended to facilitate protein or peptide ionization efficiency.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisecitation

    Dai et al. reported cIEF-MS with a sheath-liquid interface for separation of an intact monoclonal antibody, resolving nine charge variants.

    “cIEF-MS with a sheath-liquid interface has been used for the analysis of monoclonal antibodies, with Dai et al. reporting the separation of intact monoclonal antibody92. Resolving intact charge variant peaks is a noted challenge, but using this approach, nine charge variants were resolved.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [92] Dai J, Lamp J, Xia Q, Zhang Y. Capillary Isoelectric Focusing-Mass Spectrometry Method for the Separation and Online Characterization of Intact Monoclonal Antibody Charge Variants. Analytical chemistry. 2018; 90(3): 2246– 2254. 29272582 source

      Not checked in the free run.

  • May combine two claims.

    Premisereasoning

    Neutral polyacrylamide coating is widely used in proteomics because it eliminates EOF and maintains effective CE separations.

    “Neutral coating with polyacrylamide is widely used in proteomics research due to its ability to eliminate EOF and maintain effective CE separations.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: Coated capillaries can limit protein adsorption on the capillary wall.
    • Rests on an assumption: EOF suppression or reversal can result in higher separation efficiency.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Interpretationreasoning

    The high charge states seen with ammonium bicarbonate may result from bicarbonate anion destabilizing the protein and increasing its unfolding during ESI.

    “This phenomenon might be attributed to the destabilization effects that are induced by the anion bicarbonate, exacerbating the propensity of the protein to unfold during ESI process.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Premisenone given

    Many preconcentration techniques for CE-MS enhance injection volumes and are based on electrophoretic stacking or chromatographic solid-phase extraction principles.

    “Many preconcentration techniques to enhance injection volumes in CE-MS applications have been described, based on either electrophoretic (stacking) or chromatographic (solid-phase extraction (SPE)) principles.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premiseown data

    Acetate, bicarbonate, and formate salts each have more than one pKa value, so they can be used over more than one pH range.

    “As can be observed in Table 2, these salts (acetate, bicarbonate, and formate) have more than one pKa, which allow their use in more than one pH range.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    A CE-ESI-MS sheathless interface with ammonium bicarbonate preserved the native structure of Beta 2-microglobulin conformers and did not produce the high charge states promoted by ammonium bicarbonate, possibly because of lower ammonium bicarbonate concentration in the BGE.

    “Using this set up, Bertoletti et al. were able to preserve protein structure in native conditions and they did not observe the high charge states that are promoted by ammonium bicarbonate, which might be attributed to the lower molar concentration of ammonium bicarbonate in the BGE.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [102] Bertoletti L, Schappler J, Colombo R, Evaluation of capillary electrophoresis-mass spectrometry for the analysis of the conformational heterogeneity of intact proteins using beta2-microglobulin as model compound. Analytica chimica acta. 2016; 945: 102– 109. 27968711 source

      Not checked in the free run.

  • May combine two claims.

    Premisecitation

    A two-plex CE-MS method for N-glycan quantification was linear and precise.

    “The CE-MS method was found to be linear and precise.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [128] Varadi C, Mittermayr S, Millan-Martin S, Bones J. Quantitative twoplex glycan analysis using (12)C6 and (13)C6 stable isotope 2-aminobenzoic acid labelling and capillary electrophoresis mass spectrometry. Analytical and bioanalytical chemistry. 2016; 408(30): 8691– 8700. 27662881 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    Despite more technical difficulties than LC-MS, CE-MS with improved instrumentation has outperformed conventional LC-based methods in some applications, such as separation of intact proteins and protein complexes.

    “Despite susceptibility to more technical difficulties and issues than LC-MS, with improvements in instrumentation CE-MS has, in some applications, outperformed conventional LC-based methods (e.g., separation of intact proteins and protein complexes).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationnone given

    Both sheathless and sheath-liquid interfaces can achieve highly sensitive ionization at lower flow-rate than LC, reducing background noise and increasing MS detectability.

    “Highly sensitive ionization efficiency can be achieved using both sheathless and sheath-liquid interfaces at lower flow-rate (in the separation capillary) than LC, which reduces background noise and increases MS detectability.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    CE-MS characterized and quantitated individual molecular proteoforms of interferon-β.

    “Bush et al. characterized and quantitated individual molecular proteoforms of interferon-β111.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [111] Sanchez-Hernandez L, Montealegre C, Kiessig S, Moritz B, Neususs C. In-capillary approach to eliminate SDS interferences in antibody analysis by capillary electrophoresis coupled to mass spectrometry. Electrophoresis. 2017; 38(7): 1044– 1052. 28008632 source

      Not checked in the free run.

  • May combine two claims.

    Interpretationreasoning

    SPE-CE technology can be automated, and multistep elution CE can serve as a semi-two-dimensional separation method.

    “This technology has the advantage that it can be automated and the multistep elution CE can be used as a semi-two-dimensional separation method.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premisecitation

    On-line SPE preconcentration with a sulfonate-silica hybrid SCX monolith and dynamic pH junction enabled microliter-level injections with separation efficiency over 25,000 theoretical plates.

    “This approach enabled injections at μL levels with a separation efficiency of over 25,000 theoretical plates.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [124] Zhang Z, Sun L, Zhu G, Yan X, Dovichi NJ. Integrated strong cation-exchange hybrid monolith coupled with capillary zone electrophoresis and simultaneous dynamic pH junction for large-volume proteomic analysis by mass spectrometry. Talanta. 2015; 138: 117– 122. 25863379 PMC4394190 source

      Not checked in the free run.

    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premisenone given

    TDMS can provide structural information on proteins covalently attached to other proteins, while native MS provides information on protein subunit arrangements via non-covalent interactions.

    “In addition to a detailed proteoform characterization, TDMS can provide structural information of proteins covalently attached to other proteins while native MS provides a detailed information on protein subunit arrangements via non-covalent interactions.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Preparing SCX and a neutral coated capillary sequentially in a single fused silica capillary eliminated the zero dead volume union and enhanced protein and peptide identifications compared to previous reports.

    “The same group further eliminated the zero dead volume union by preparing SCX and the neutral coated capillary sequentially in a single fused silica capillary, which resulted in an enhanced protein and peptide identifications when compared to previous reports126.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [126] Zhang Z, Zhu G, Peuchen EH, Dovichi NJ. Preparation of linear polyacrylamide coating and strong cationic exchange hybrid monolith in a single capillary, and its application as an automated platform for bottom-up proteomics by capillary electrophoresis-mass spectrometry. Microchimica Acta. 2017; 184(3): 921– 925. source

      Not checked in the free run.

    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premisecitation

    cITP/CZE quantification of targeted BSA tryptic peptides had a quantitation limit of 10 pM and a coefficient of variation lower than 22%.

    “They observed a quantitation limit of 10 pM with a CV lower than 22%121.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [121] Wang C, Lee CS, Smith RD, Tang KJAc. CITP/CZE-nanoESI-SRM MS via a novel sheathless interface for high sensitivity sample quantification. 2013; 85(15): 7308. PMC3744340 23789856 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Interpretationnone given

    Cross-linking and MS have been successfully used to interrogate protein structures, but CE-MS cross-linking reports are scarce.

    “Cross-linking and MS have been successfully used to interrogate protein structures; however, cross-linking reports by CE-MS are scarce.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationcitation

    Sample loading is the major drawback of CE and can be addressed using simple protocols, as shown in top-down and bottom-up applications.

    “Sample loading is the major drawback of CE, which can be addressed using simple protocols, as demonstrated in top-down and bottom-up applications52,87.”
    Quote in the paperSubstantial claim0 of 2 sources checked
    • cites [52] McCool EN, Lubeckyj R, Shen X, Kou Q, Liu X, Sun L. Large-scale Top-down Proteomics Using Capillary Zone Electrophoresis Tandem Mass Spectrometry. Journal of visualized experiments: JoVE. 2018(140). PMC6235596 30417888 source

      Not checked in the free run.

    • cites [87] Wang Y, Fonslow BR, Wong CC, Nakorchevsky A, Yates JR 3rd. Improving the comprehensiveness and sensitivity of sheathless capillary electrophoresis-tandem mass spectrometry for proteomic analysis. Analytical chemistry. 2012; 84(20): 8505– 8513. 23004022 PMC3498465 source

      Not checked in the free run.

    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisenone given

    tITP mixes the sample with high-mobility leading and low-mobility terminating electrolytes, generating a highly conductive zone when voltage is applied and concentrating proteins or peptides between the leading and terminating electrolytes.

    “In this strategy, the sample is mixed with two electrolytes, one with very high mobility (leading) and one with very low mobility (terminating). These electrolytes generate a highly conductive zone when separation voltage is applied. Proteins or peptides are then concentrated into zones between the leading and terminating electrolytes.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Premisecitation

    Reversed-phase SPE combined with tITP for a Pyrococcus furiosus tryptic digest gave low background noise and high sensitivity.

    “Wang et al. used reversed-phase SPE in conjunction with tITP for sample concentration of a Pyrococcus furiosus tryptic digest, which resulted in low background noise and high sensitivity87.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [87] Wang Y, Fonslow BR, Wong CC, Nakorchevsky A, Yates JR 3rd. Improving the comprehensiveness and sensitivity of sheathless capillary electrophoresis-tandem mass spectrometry for proteomic analysis. Analytical chemistry. 2012; 84(20): 8505– 8513. 23004022 PMC3498465 source

      Not checked in the free run.

    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisenone given

    Several bottom-up strategies are available for protein quantification, including SILAC, iTRAQ, label-free, and ICAT.

    “Several bottom-up strategies are available for protein quantification, including stable isotope labeling by amino acids in cell culture (SILAC), isobaric tag for relative and absolute quantitation (iTRAQ), label-free, and isotope-coded affinity tags (ICAT).”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationreasoning

    Improvements in CE-MS interfaces have enhanced robustness and reproducibility, leading to confident identification and characterization of peptides, cellular proteins, PTMs, and protein complexes.

    “Improvements in interfaces have enhanced the robustness and reproducibility of CE-MS methods, resulting in highly confident identification and characterization of peptides, cellular proteins, PTMs, and protein complexes.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Premisecitation

    Quantitative parallel reaction monitoring of peptide abundance had a relative standard deviation of migration time lower than 4% and a relative standard deviation of fragment ion of approximately 20%.

    “Sun and co-workers described a quantitative parallel reaction monitoring of peptide abundance where the relative standard deviation (RSD) of the migration time was lower than 4% and the RSD of the fragment ion was approximately 20%132.”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [132] Sun L, Zhu G, Mou S, Zhao Y, Champion MM, Dovichi NJ. Capillary zone electrophoresis-electrospray ionization-tandem mass spectrometry for quantitative parallel reaction monitoring of peptide abundance and single-shot proteomic analysis of a human cell line. Journal of chromatography A. 2014; 1359: 303– 308. 25082526 PMC4141985 source

      Not checked in the free run.

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisenone given

    Transient isotachophoresis is a less common stacking approach for preconcentration of peptides and proteins in proteomics.

    “Transient isotachophoresis (tITP) is a less common stacking approach for preconcentration of peptides and proteins in proteomics research.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

  • May combine two claims.

    Interpretationreasoning

    TDMS and native MS can be complementary at several levels and merged to provide comprehensive protein structural information.

    “These individual features suggest that TDMS and native MS can be complementary at several levels and can be merged to provide comprehensive protein structural information.”
    Quote in the paperSubstantial claim
70 of 70

  • The paper relies on this without support.

    Progress in proteomics research has created a demand for powerful analytical tools with high separation efficiency and sensitivity for confident identification and quantification of proteins, PTMs, and protein complexes in cells and tissues.

    “Progress in proteomics research has led to a demand for powerful analytical tools with high separation efficiency and sensitivity for confident identification and quantification of proteins, posttranslational modifications (PTMs), and protein complexes expressed in cells and tissues.”

  • The paper relies on this without support.

    This demand has significantly increased interest in CE-MS in the past few years.

    “This demand has significantly increased interest in capillary electrophoresis-mass spectrometry (CE-MS) in the past few years.”

  • The paper relies on this without support.

    The most common CE-MS technique used in proteomics is capillary zone electrophoresis coupled with electrospray ionization.

    “The most common CE-MS technique used in the proteomics field is capillary zone electrophoresis (CZE) coupled with electrospray ionization (ESI).”

  • The paper relies on this without support.

    Most proteins are assembled into protein complexes to achieve functional activity.

    “Most proteins are assembled into protein complexes to achieve functional activity, but it is still unclear how these macromolecular complexes are formed.”

  • The paper relies on this without support.

    Residue shape and size have little effect on the electrophoretic mobility of a peptide.

    “Since it has been generally accepted that residue shape and size have little effect on the electrophoretic mobility of a peptide, these models are based on the use of BGE pH, side group pKa, N-terminal amine, and C-terminal acid to determine the charge of the peptide.”

  • The paper relies on this without support.

    In CZE, band broadening is caused only by longitudinal diffusion.

    “In CZE, band broadening is only caused by longitudinal diffusion.”

  • The paper relies on this without support.

    CE has very limited loading capacity, which can reduce sensitivity and peptide and protein identifications.

    “The loading capacity of CE is very limited, which can adversely affect the sensitivity of the method and reduce peptide and protein identifications.”

  • The paper relies on this without support.

    Proteins have low diffusion coefficients and should therefore produce very high separation efficiency in CE.

    “Theoretically, since proteins have low diffusion coefficients, they should produce very high separation efficiency in CE.”

  • The paper relies on this without support.

    Hydrophobic and/or ionic interactions between proteins and capillary wall surfaces can increase protein retention or cause peak tailing.

    “However, hydrophobic and/or ionic interactions between protein and capillary wall surfaces can lead to increased protein retention in the capillary, or to a lesser extent, peak tailing.”

  • The paper relies on this without support.

    Coated capillaries can limit protein adsorption on the capillary wall.

    “Coated capillaries can be used to limit protein adsorption on the capillary wall.”

  • The paper relies on this without support.

    EOF suppression or reversal can result in higher separation efficiency.

    “EOF suppression or reversal can result in higher separation efficiency, and the EOF can be regulated by the capillary coating.”

  • The paper relies on this without support.

    In native ESI-MS, environmentally induced pH changes can perturb tertiary and quaternary conformations of protein complexes in solution.

    “In native ESI-MS, protein complexes are transferred from solution to the gas phase and environmentally induced changes in pH can perturb their tertiary and quaternary conformations while in solution.”

  • The paper relies on this without support.

    High ionic strength can increase diffusion of the sample zone and lead to high current and temperature in the separation capillary.

    “Although a BGE with high ionic strength is useful to avoid band broadening, high ionic strength can increase the diffusion of the sample zone and lead to high current and temperature in the separation capillary.”

  • The paper relies on this without support.

    Quantitative information is critical to gain insights into clinical disorders.

    “In proteomics research, quantitative information is critical to gain insights into clinical disorders.”

  • The paper relies on this without support.

    Quantification via CE-MS is particularly valuable for clinical studies where samples are often obtained in limited quantities.

    “Quantification via CE-MS is particularly valuable for clinical studies where samples are often obtained in limited quantities.”

  • The paper relies on this without support.

    Top-down analysis provides quantitative information on diverse proteoforms that are often missed in bottom-up approaches.

    “Top-down analysis provides quantitative information on diverse proteoforms that are often missed in bottom-up approaches.”

  • The paper relies on this without support.

    No single dimension can provide complete reduction of sample complexity.

    “No single dimension, however, can provide complete reduction of sample complexity.”

  • The paper relies on this without support.

    Multidimensional separations are the most effective strategy for improving peak capacity and reducing sample complexity.

    “Rather, utilization of multidimensional separations is the most effective strategy for improving peak capacity and reducing sample complexity.”
18 of 18

Claims other papers make about this one

Each is checked against this paper’s own text: does it say what the citing paper says it does?

Self-check

Percentages, sums and reported differences re-computed from the paper’s own numbers.

Each contradiction between two passages listed here was confirmed by an independent check.

  • Belov study is attributed to a reference by different authors

    Minor

    The text attributes reference 34 to Belov and co-workers, but reference 34 lists Chen and co-workers and describes a different study.

  • SPE study is attributed to Wang instead of the cited authors

    Minor

    The text names Wang et al. for the study cited as reference 123, but that reference lists Tong et al.

2 of 2

Cites

Flagged first, then by citations

Cited by

Sorted by citations

Related

Authors
Gomes, F. P., & Yates, J. R.
Affiliations
  • The Scripps Research Institute La Jolla CA 92037
  • Scripps Research Institute (US)
Corresponding author
Yates, John R.
ORCID
Yates, John R.
Venue
Mass Spectrometry Reviews
Publisher
Wiley
Published
2019-08-12
Volume
38
Issue
6
Pages
445-460
ISSN
  • 0277-7037 (ISSN-L)
  • 1098-2787
DOI
10.1002/mas.21599
PMC
PMC6800771
OpenAlex
W2968805832
Cited by
112
References
141
Citations by year
2026: 8 · 2025: 15 · 2024: 13 · 2023: 17 · 2022: 20 · 2021: 23 · 2020: 15 · 2019: 1
FWCI
1.53
Type
journal article
Language
en
Open access
Licence
onlinelibrary.wiley.com/termsAndConditions#vor (vor)
Locations
  • Mass Spectrometry Reviews (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • National Institutes of Health
  • National Institute of General Medical Sciences (P41GM103533)
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
  • Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
  • Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
Keywords
capillary electrophoresis-mass spectrometry, proteomics, top-down mass spectrometry, native mass spectrometry, protein identification, protein quantification, post-translational modification, protein complexes, peptide analysis, biological specimen
MeSH terms
Animals, Humans, Mass Spectrometry, Electrophoresis, Capillary, Proteome, Spectrometry, Mass, Electrospray Ionization, Proteomics

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-05.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-05.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review