Quantitation of peptides from non-invasive skin tapings using isotope dilution and tandem mass spectrometry.
Identifieur interne : 000669 ( Main/Exploration ); précédent : 000668; suivant : 000670Quantitation of peptides from non-invasive skin tapings using isotope dilution and tandem mass spectrometry.
Auteurs : Nichole Reisdorph [États-Unis] ; Michael Armstrong [États-Unis] ; Roger Powell [États-Unis] ; Kevin Quinn [États-Unis] ; Kevin Legg [États-Unis] ; Donald Leung [États-Unis] ; Rick Reisdorph [États-Unis]Source :
- Journal of chromatography. B, Analytical technologies in the biomedical and life sciences [ 1873-376X ] ; 2018.
Descripteurs français
- KwdFr :
- Eczéma atopique (métabolisme), Humains (MeSH), Limite de détection (MeSH), Marquage isotopique (méthodes), Modèles linéaires (MeSH), Peau (composition chimique), Peptides (analyse), Protéomique (méthodes), Reproductibilité des résultats (MeSH), Spectrométrie de masse en tandem (méthodes), Études cas-témoins (MeSH).
- MESH :
- analyse : Peptides.
- composition chimique : Peau.
- métabolisme : Eczéma atopique.
- méthodes : Marquage isotopique, Protéomique, Spectrométrie de masse en tandem.
- Humains, Limite de détection, Modèles linéaires, Reproductibilité des résultats, Études cas-témoins.
English descriptors
- KwdEn :
- MESH :
- chemical , analysis : Peptides.
- chemistry : Skin.
- metabolism : Dermatitis, Atopic.
- methods : Isotope Labeling, Proteomics, Tandem Mass Spectrometry.
- Case-Control Studies, Humans, Limit of Detection, Linear Models, Reproducibility of Results.
Abstract
Previous work from our laboratories utilized a novel skin taping method and mass spectrometry-based proteomics to discover clinical biomarkers of skin conditions; these included atopic dermatitis, Staphylococcus aureus colonization, and eczema herpeticum. While suitable for discovery purposes, semi-quantitative proteomics is generally time-consuming and expensive. Furthermore, depending on the method used, discovery-based proteomics can result in high variation and inadequate sensitivity to detect low abundant peptides. Therefore, we strove to develop a rapid, sensitive, and reproducible method to quantitate disease-related proteins from skin tapings. We utilized isotopically-labeled peptides and tandem mass spectrometry to obtain absolute quantitation values on 14 peptides from 7 proteins; these proteins had shown previous importance in skin disease. The method demonstrated good reproducibility, dynamic range, and linearity (R2 > 0.993) when n = 3 standards were analyzed across 0.05-2.5 pmol. The method was used to determine if differences exist between skin proteins in a small group of atopic versus non-atopic individuals (n = 12). While only minimal differences were found, peptides were detected in all samples and exhibited good correlation between peptides for 5 of the 7 proteins (R2 = 0.71-0.98). This method can be applied to larger cohorts to further establish the relationships of these proteins to skin disease.
DOI: 10.1016/j.jchromb.2018.03.031
PubMed: 29601982
PubMed Central: PMC6093185
Affiliations:
Links toward previous steps (curation, corpus...)
Le document en format XML
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<term>Isotope Labeling (methods)</term>
<term>Limit of Detection (MeSH)</term>
<term>Linear Models (MeSH)</term>
<term>Peptides (analysis)</term>
<term>Proteomics (methods)</term>
<term>Reproducibility of Results (MeSH)</term>
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<term>Tandem Mass Spectrometry (methods)</term>
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<keywords scheme="KwdFr" xml:lang="fr"><term>Eczéma atopique (métabolisme)</term>
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<term>Limite de détection (MeSH)</term>
<term>Marquage isotopique (méthodes)</term>
<term>Modèles linéaires (MeSH)</term>
<term>Peau (composition chimique)</term>
<term>Peptides (analyse)</term>
<term>Protéomique (méthodes)</term>
<term>Reproductibilité des résultats (MeSH)</term>
<term>Spectrométrie de masse en tandem (méthodes)</term>
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<front><div type="abstract" xml:lang="en">Previous work from our laboratories utilized a novel skin taping method and mass spectrometry-based proteomics to discover clinical biomarkers of skin conditions; these included atopic dermatitis, Staphylococcus aureus colonization, and eczema herpeticum. While suitable for discovery purposes, semi-quantitative proteomics is generally time-consuming and expensive. Furthermore, depending on the method used, discovery-based proteomics can result in high variation and inadequate sensitivity to detect low abundant peptides. Therefore, we strove to develop a rapid, sensitive, and reproducible method to quantitate disease-related proteins from skin tapings. We utilized isotopically-labeled peptides and tandem mass spectrometry to obtain absolute quantitation values on 14 peptides from 7 proteins; these proteins had shown previous importance in skin disease. The method demonstrated good reproducibility, dynamic range, and linearity (R<sup>2</sup>
> 0.993) when n = 3 standards were analyzed across 0.05-2.5 pmol. The method was used to determine if differences exist between skin proteins in a small group of atopic versus non-atopic individuals (n = 12). While only minimal differences were found, peptides were detected in all samples and exhibited good correlation between peptides for 5 of the 7 proteins (R<sup>2</sup>
= 0.71-0.98). This method can be applied to larger cohorts to further establish the relationships of these proteins to skin disease.</div>
</front>
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<Abstract><AbstractText>Previous work from our laboratories utilized a novel skin taping method and mass spectrometry-based proteomics to discover clinical biomarkers of skin conditions; these included atopic dermatitis, Staphylococcus aureus colonization, and eczema herpeticum. While suitable for discovery purposes, semi-quantitative proteomics is generally time-consuming and expensive. Furthermore, depending on the method used, discovery-based proteomics can result in high variation and inadequate sensitivity to detect low abundant peptides. Therefore, we strove to develop a rapid, sensitive, and reproducible method to quantitate disease-related proteins from skin tapings. We utilized isotopically-labeled peptides and tandem mass spectrometry to obtain absolute quantitation values on 14 peptides from 7 proteins; these proteins had shown previous importance in skin disease. The method demonstrated good reproducibility, dynamic range, and linearity (R<sup>2</sup>
> 0.993) when n = 3 standards were analyzed across 0.05-2.5 pmol. The method was used to determine if differences exist between skin proteins in a small group of atopic versus non-atopic individuals (n = 12). While only minimal differences were found, peptides were detected in all samples and exhibited good correlation between peptides for 5 of the 7 proteins (R<sup>2</sup>
= 0.71-0.98). This method can be applied to larger cohorts to further establish the relationships of these proteins to skin disease.</AbstractText>
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