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Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS
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| Zeitschriftentitel: | Journal of Cellular and Molecular Medicine |
|---|---|
| Personen und Körperschaften: | , , , , , |
| In: | Journal of Cellular and Molecular Medicine, 19, 2015, 7, S. 1656-1671 |
| Format: | E-Article |
| Sprache: | Englisch |
| veröffentlicht: |
Wiley
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| Schlagwörter: |
| author_facet |
Lancrajan, Ioana Schneider‐Stock, Regine Naschberger, Elisabeth Schellerer, Vera S. Stürzl, Michael Enz, Ralf Lancrajan, Ioana Schneider‐Stock, Regine Naschberger, Elisabeth Schellerer, Vera S. Stürzl, Michael Enz, Ralf |
|---|---|
| author |
Lancrajan, Ioana Schneider‐Stock, Regine Naschberger, Elisabeth Schellerer, Vera S. Stürzl, Michael Enz, Ralf |
| spellingShingle |
Lancrajan, Ioana Schneider‐Stock, Regine Naschberger, Elisabeth Schellerer, Vera S. Stürzl, Michael Enz, Ralf Journal of Cellular and Molecular Medicine Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS Cell Biology Molecular Medicine |
| author_sort |
lancrajan, ioana |
| spelling |
Lancrajan, Ioana Schneider‐Stock, Regine Naschberger, Elisabeth Schellerer, Vera S. Stürzl, Michael Enz, Ralf 1582-1838 1582-4934 Wiley Cell Biology Molecular Medicine http://dx.doi.org/10.1111/jcmm.12540 <jats:title>Abstract</jats:title><jats:p>Biomarkers are widely used in clinical diagnosis, prognosis and therapy monitoring. Here, we developed a protocol for the efficient and selective enrichment of small and low concentrated biomarkers from human serum, involving a 95% effective depletion of high‐abundant serum proteins by partial denaturation and enrichment of low‐abundant biomarkers by size exclusion chromatography. The recovery of low‐abundance biomarkers was above 97%. Using this protocol, we quantified the tumour markers DcR3 and growth/differentiation factor (<jats:styled-content style="fixed-case">GDF</jats:styled-content>)15 from 100 μl human serum by isotope dilution mass spectrometry, using <jats:sup>15</jats:sup>N metabolically labelled and concatamerized fingerprint peptides for the both proteins. Analysis of three different fingerprint peptides for each protein by liquid chromatography electrospray ionization mass spectrometry resulted in comparable concentrations in three healthy human serum samples (DcR3: 27.23 ± 2.49 fmol/ml; <jats:styled-content style="fixed-case">GDF</jats:styled-content>15: 98.11 ± 0.49 fmol/ml). In contrast, serum levels were significantly elevated in tumour patients for DcR3 (116.94 ± 57.37 fmol/ml) and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 (164.44 ± 79.31 fmol/ml). Obtained data were in good agreement with <jats:styled-content style="fixed-case">ELISA</jats:styled-content> and <jats:styled-content style="fixed-case">qPCR</jats:styled-content> measurements, as well as with literature data. In summary, our protocol allows the reliable quantification of biomarkers, shows a higher resolution at low biomarker concentrations than antibody‐based strategies, and offers the possibility of multiplexing. Our proof‐of‐principle studies in patient sera encourage the future analysis of the prognostic value of DcR3 and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 for colon cancer patients in larger patient cohorts.</jats:p> Absolute quantification of DcR3 and <scp>GDF</scp>15 from human serum by <scp>LC</scp>‐<scp>ESI MS</scp> Journal of Cellular and Molecular Medicine |
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10.1111/jcmm.12540 |
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Wiley, 2015 |
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Wiley, 2015 |
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| title |
Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_unstemmed |
Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_full |
Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_fullStr |
Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_full_unstemmed |
Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_short |
Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_sort |
absolute quantification of dcr3 and <scp>gdf</scp>15 from human serum by <scp>lc</scp>‐<scp>esi ms</scp> |
| topic |
Cell Biology Molecular Medicine |
| url |
http://dx.doi.org/10.1111/jcmm.12540 |
| publishDate |
2015 |
| physical |
1656-1671 |
| description |
<jats:title>Abstract</jats:title><jats:p>Biomarkers are widely used in clinical diagnosis, prognosis and therapy monitoring. Here, we developed a protocol for the efficient and selective enrichment of small and low concentrated biomarkers from human serum, involving a 95% effective depletion of high‐abundant serum proteins by partial denaturation and enrichment of low‐abundant biomarkers by size exclusion chromatography. The recovery of low‐abundance biomarkers was above 97%. Using this protocol, we quantified the tumour markers DcR3 and growth/differentiation factor (<jats:styled-content style="fixed-case">GDF</jats:styled-content>)15 from 100 μl human serum by isotope dilution mass spectrometry, using <jats:sup>15</jats:sup>N metabolically labelled and concatamerized fingerprint peptides for the both proteins. Analysis of three different fingerprint peptides for each protein by liquid chromatography electrospray ionization mass spectrometry resulted in comparable concentrations in three healthy human serum samples (DcR3: 27.23 ± 2.49 fmol/ml; <jats:styled-content style="fixed-case">GDF</jats:styled-content>15: 98.11 ± 0.49 fmol/ml). In contrast, serum levels were significantly elevated in tumour patients for DcR3 (116.94 ± 57.37 fmol/ml) and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 (164.44 ± 79.31 fmol/ml). Obtained data were in good agreement with <jats:styled-content style="fixed-case">ELISA</jats:styled-content> and <jats:styled-content style="fixed-case">qPCR</jats:styled-content> measurements, as well as with literature data. In summary, our protocol allows the reliable quantification of biomarkers, shows a higher resolution at low biomarker concentrations than antibody‐based strategies, and offers the possibility of multiplexing. Our proof‐of‐principle studies in patient sera encourage the future analysis of the prognostic value of DcR3 and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 for colon cancer patients in larger patient cohorts.</jats:p> |
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| author | Lancrajan, Ioana, Schneider‐Stock, Regine, Naschberger, Elisabeth, Schellerer, Vera S., Stürzl, Michael, Enz, Ralf |
| author_facet | Lancrajan, Ioana, Schneider‐Stock, Regine, Naschberger, Elisabeth, Schellerer, Vera S., Stürzl, Michael, Enz, Ralf, Lancrajan, Ioana, Schneider‐Stock, Regine, Naschberger, Elisabeth, Schellerer, Vera S., Stürzl, Michael, Enz, Ralf |
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| description | <jats:title>Abstract</jats:title><jats:p>Biomarkers are widely used in clinical diagnosis, prognosis and therapy monitoring. Here, we developed a protocol for the efficient and selective enrichment of small and low concentrated biomarkers from human serum, involving a 95% effective depletion of high‐abundant serum proteins by partial denaturation and enrichment of low‐abundant biomarkers by size exclusion chromatography. The recovery of low‐abundance biomarkers was above 97%. Using this protocol, we quantified the tumour markers DcR3 and growth/differentiation factor (<jats:styled-content style="fixed-case">GDF</jats:styled-content>)15 from 100 μl human serum by isotope dilution mass spectrometry, using <jats:sup>15</jats:sup>N metabolically labelled and concatamerized fingerprint peptides for the both proteins. Analysis of three different fingerprint peptides for each protein by liquid chromatography electrospray ionization mass spectrometry resulted in comparable concentrations in three healthy human serum samples (DcR3: 27.23 ± 2.49 fmol/ml; <jats:styled-content style="fixed-case">GDF</jats:styled-content>15: 98.11 ± 0.49 fmol/ml). In contrast, serum levels were significantly elevated in tumour patients for DcR3 (116.94 ± 57.37 fmol/ml) and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 (164.44 ± 79.31 fmol/ml). Obtained data were in good agreement with <jats:styled-content style="fixed-case">ELISA</jats:styled-content> and <jats:styled-content style="fixed-case">qPCR</jats:styled-content> measurements, as well as with literature data. In summary, our protocol allows the reliable quantification of biomarkers, shows a higher resolution at low biomarker concentrations than antibody‐based strategies, and offers the possibility of multiplexing. Our proof‐of‐principle studies in patient sera encourage the future analysis of the prognostic value of DcR3 and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 for colon cancer patients in larger patient cohorts.</jats:p> |
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| imprint | Wiley, 2015 |
| imprint_str_mv | Wiley, 2015 |
| institution | DE-D275, DE-Bn3, DE-Brt1, DE-Zwi2, DE-D161, DE-Zi4, DE-Gla1, DE-15, DE-Pl11, DE-Rs1, DE-14, DE-105, DE-Ch1, DE-L229 |
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| spelling | Lancrajan, Ioana Schneider‐Stock, Regine Naschberger, Elisabeth Schellerer, Vera S. Stürzl, Michael Enz, Ralf 1582-1838 1582-4934 Wiley Cell Biology Molecular Medicine http://dx.doi.org/10.1111/jcmm.12540 <jats:title>Abstract</jats:title><jats:p>Biomarkers are widely used in clinical diagnosis, prognosis and therapy monitoring. Here, we developed a protocol for the efficient and selective enrichment of small and low concentrated biomarkers from human serum, involving a 95% effective depletion of high‐abundant serum proteins by partial denaturation and enrichment of low‐abundant biomarkers by size exclusion chromatography. The recovery of low‐abundance biomarkers was above 97%. Using this protocol, we quantified the tumour markers DcR3 and growth/differentiation factor (<jats:styled-content style="fixed-case">GDF</jats:styled-content>)15 from 100 μl human serum by isotope dilution mass spectrometry, using <jats:sup>15</jats:sup>N metabolically labelled and concatamerized fingerprint peptides for the both proteins. Analysis of three different fingerprint peptides for each protein by liquid chromatography electrospray ionization mass spectrometry resulted in comparable concentrations in three healthy human serum samples (DcR3: 27.23 ± 2.49 fmol/ml; <jats:styled-content style="fixed-case">GDF</jats:styled-content>15: 98.11 ± 0.49 fmol/ml). In contrast, serum levels were significantly elevated in tumour patients for DcR3 (116.94 ± 57.37 fmol/ml) and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 (164.44 ± 79.31 fmol/ml). Obtained data were in good agreement with <jats:styled-content style="fixed-case">ELISA</jats:styled-content> and <jats:styled-content style="fixed-case">qPCR</jats:styled-content> measurements, as well as with literature data. In summary, our protocol allows the reliable quantification of biomarkers, shows a higher resolution at low biomarker concentrations than antibody‐based strategies, and offers the possibility of multiplexing. Our proof‐of‐principle studies in patient sera encourage the future analysis of the prognostic value of DcR3 and <jats:styled-content style="fixed-case">GDF</jats:styled-content>15 for colon cancer patients in larger patient cohorts.</jats:p> Absolute quantification of DcR3 and <scp>GDF</scp>15 from human serum by <scp>LC</scp>‐<scp>ESI MS</scp> Journal of Cellular and Molecular Medicine |
| spellingShingle | Lancrajan, Ioana, Schneider‐Stock, Regine, Naschberger, Elisabeth, Schellerer, Vera S., Stürzl, Michael, Enz, Ralf, Journal of Cellular and Molecular Medicine, Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS, Cell Biology, Molecular Medicine |
| title | Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_full | Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_fullStr | Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_full_unstemmed | Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_short | Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| title_sort | absolute quantification of dcr3 and <scp>gdf</scp>15 from human serum by <scp>lc</scp>‐<scp>esi ms</scp> |
| title_unstemmed | Absolute quantification of DcR3 and GDF15 from human serum by LC‐ESI MS |
| topic | Cell Biology, Molecular Medicine |
| url | http://dx.doi.org/10.1111/jcmm.12540 |