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- Publisher Website: 10.1002/jms.4591
- Scopus: eid_2-s2.0-85087627835
- PMID: 32633895
- WOS: WOS:000545759500001
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Article: Implementation of a multiple-fraction concatenation strategy in an online two-dimensional high-/low-pH reversed-phase/reversed-phase liquid chromatography platform for qualitative and quantitative shotgun proteomic analyses
Title | Implementation of a multiple-fraction concatenation strategy in an online two-dimensional high-/low-pH reversed-phase/reversed-phase liquid chromatography platform for qualitative and quantitative shotgun proteomic analyses |
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Authors | |
Keywords | Peptides Separation Critical factors Multidimensional liquid chromatography Proteomic analysis |
Issue Date | 2020 |
Publisher | John Wiley & Sons Ltd. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/6043 |
Citation | Journal of Mass Spectrometry, 2020, Epub 2020-06-12, p. article no. e4591 How to Cite? |
Abstract | Multidimensional liquid chromatography is the mainstay separation technique used for shotgun proteomic analyses. The application of a multiple‐fraction concatenation (MFC) strategy can result in a more disperse and consistent peptide elution profile across different fractions, when compared with a conventional strategy. Herein, we present the first automated online RP‐RP platform implementing an MFC strategy to facilitate robust, unattended, routine proteomic analyses. The improved duty cycle utilization of the MFC strategy led to an increase of 9% in the separation space occupancy and increases of approximately 10% in the identification of both proteins and peptides. The peptides uniquely identified by the MFC strategy were significantly biased toward those of acidic nature, with increased precursor signals leading to improved MS/MS spectral quality and enhanced acidic peptide identification. These improvements in qualitative analysis using the MFC strategy were also extended to quantitative analysis. When the acquired proteome was quantified with a normalized spectral abundance factor, the additionally acquired acidic peptides were a critical factor leading to enhanced reproducibility of quantitation using the MFC strategy. With merits of superior qualitative and quantitative characteristics over the conventional strategy, the MFC strategy appears to be a highly amenable technique for enhancing the separation capacity for routine proteomic analyses. |
Persistent Identifier | http://hdl.handle.net/10722/284016 |
ISSN | 2023 Impact Factor: 1.9 2023 SCImago Journal Rankings: 0.431 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Law, CH | - |
dc.contributor.author | Kong, R | - |
dc.contributor.author | Li, MZ | - |
dc.contributor.author | Szeto, SSW | - |
dc.contributor.author | Chu, IK | - |
dc.date.accessioned | 2020-07-20T05:55:21Z | - |
dc.date.available | 2020-07-20T05:55:21Z | - |
dc.date.issued | 2020 | - |
dc.identifier.citation | Journal of Mass Spectrometry, 2020, Epub 2020-06-12, p. article no. e4591 | - |
dc.identifier.issn | 1076-5174 | - |
dc.identifier.uri | http://hdl.handle.net/10722/284016 | - |
dc.description.abstract | Multidimensional liquid chromatography is the mainstay separation technique used for shotgun proteomic analyses. The application of a multiple‐fraction concatenation (MFC) strategy can result in a more disperse and consistent peptide elution profile across different fractions, when compared with a conventional strategy. Herein, we present the first automated online RP‐RP platform implementing an MFC strategy to facilitate robust, unattended, routine proteomic analyses. The improved duty cycle utilization of the MFC strategy led to an increase of 9% in the separation space occupancy and increases of approximately 10% in the identification of both proteins and peptides. The peptides uniquely identified by the MFC strategy were significantly biased toward those of acidic nature, with increased precursor signals leading to improved MS/MS spectral quality and enhanced acidic peptide identification. These improvements in qualitative analysis using the MFC strategy were also extended to quantitative analysis. When the acquired proteome was quantified with a normalized spectral abundance factor, the additionally acquired acidic peptides were a critical factor leading to enhanced reproducibility of quantitation using the MFC strategy. With merits of superior qualitative and quantitative characteristics over the conventional strategy, the MFC strategy appears to be a highly amenable technique for enhancing the separation capacity for routine proteomic analyses. | - |
dc.language | eng | - |
dc.publisher | John Wiley & Sons Ltd. The Journal's web site is located at http://www3.interscience.wiley.com/cgi-bin/jhome/6043 | - |
dc.relation.ispartof | Journal of Mass Spectrometry | - |
dc.rights | Preprint This is the pre-peer reviewed version of the following article: [FULL CITE], which has been published in final form at [Link to final article using the DOI]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. Postprint This is the peer reviewed version of the following article: [FULL CITE], which has been published in final form at [Link to final article using the DOI]. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions. | - |
dc.subject | Peptides | - |
dc.subject | Separation | - |
dc.subject | Critical factors | - |
dc.subject | Multidimensional liquid chromatography | - |
dc.subject | Proteomic analysis | - |
dc.title | Implementation of a multiple-fraction concatenation strategy in an online two-dimensional high-/low-pH reversed-phase/reversed-phase liquid chromatography platform for qualitative and quantitative shotgun proteomic analyses | - |
dc.type | Article | - |
dc.identifier.email | Chu, IK: ivankchu@hkucc.hku.hk | - |
dc.identifier.authority | Chu, IK=rp00683 | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1002/jms.4591 | - |
dc.identifier.pmid | 32633895 | - |
dc.identifier.scopus | eid_2-s2.0-85087627835 | - |
dc.identifier.hkuros | 311055 | - |
dc.identifier.volume | Epub 2020-06-12 | - |
dc.identifier.spage | article no. e4591 | - |
dc.identifier.epage | article no. e4591 | - |
dc.identifier.isi | WOS:000545759500001 | - |
dc.publisher.place | United Kingdom | - |
dc.identifier.issnl | 1076-5174 | - |