Comparison of different depletion strategies for improving resolution of the human urine proteome
Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins,...
Ausführliche Beschreibung
Gespeichert in:
| Autor*in: |
|---|
| Format: |
E-Artikel |
|---|
| Erschienen: |
Walter de Gruyter ; 2010 |
|---|
| Schlagwörter: |
|---|
| Umfang: |
5 |
|---|
| Reproduktion: |
Walter de Gruyter Online Zeitschriften |
|---|---|
| Übergeordnetes Werk: |
Enthalten in: Clinical chemistry and laboratory medicine - Berlin [u.a.] : De Gruyter, 1998, 48(2010), 4 vom: 12. Feb., Seite 531-535 |
| Übergeordnetes Werk: |
volume:48 ; year:2010 ; number:4 ; day:12 ; month:02 ; pages:531-535 ; extent:5 |
| Links: |
|---|
| DOI / URN: |
10.1515/CCLM.2010.109 |
|---|
| Katalog-ID: |
NLEJ246730161 |
|---|
| LEADER | 01000caa a22002652 4500 | ||
|---|---|---|---|
| 001 | NLEJ246730161 | ||
| 003 | DE-627 | ||
| 005 | 20230506145551.0 | ||
| 007 | cr uuu---uuuuu | ||
| 008 | 220814s2010 xx |||||o 00| ||und c | ||
| 024 | 7 | |a 10.1515/CCLM.2010.109 |2 doi | |
| 028 | 5 | 2 | |a artikel_Grundlieferung.pp |
| 035 | |a (DE-627)NLEJ246730161 | ||
| 040 | |a DE-627 |b ger |c DE-627 |e rakwb | ||
| 245 | 1 | 0 | |a Comparison of different depletion strategies for improving resolution of the human urine proteome |
| 264 | 1 | |b Walter de Gruyter |c 2010 | |
| 300 | |a 5 | ||
| 336 | |a Text |b txt |2 rdacontent | ||
| 337 | |a Computermedien |b c |2 rdamedia | ||
| 338 | |a Online-Ressource |b cr |2 rdacarrier | ||
| 520 | |a Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. | ||
| 533 | |f Walter de Gruyter Online Zeitschriften | ||
| 650 | 4 | |a biomarkers | |
| 650 | 4 | |a proteins depletion | |
| 650 | 4 | |a proteomics | |
| 650 | 4 | |a urine | |
| 700 | 1 | |a Magagnotti, Cinzia |4 oth | |
| 700 | 1 | |a Fermo, Isabella |4 oth | |
| 700 | 1 | |a Carletti, Rose Mary |4 oth | |
| 700 | 1 | |a Ferrari, Maurizio |4 oth | |
| 700 | 1 | |a Bachi, Angela |4 oth | |
| 773 | 0 | 8 | |i Enthalten in |t Clinical chemistry and laboratory medicine |d Berlin [u.a.] : De Gruyter, 1998 |g 48(2010), 4 vom: 12. Feb., Seite 531-535 |w (DE-627)NLEJ248235222 |w (DE-600)1492732-9 |x 1437-4331 |7 nnns |
| 773 | 1 | 8 | |g volume:48 |g year:2010 |g number:4 |g day:12 |g month:02 |g pages:531-535 |g extent:5 |
| 856 | 4 | 0 | |u https://doi.org/10.1515/CCLM.2010.109 |z Deutschlandweit zugänglich |
| 912 | |a GBV_USEFLAG_U | ||
| 912 | |a ZDB-1-DGR | ||
| 912 | |a GBV_NL_ARTICLE | ||
| 951 | |a AR | ||
| 952 | |d 48 |j 2010 |e 4 |b 12 |c 02 |h 531-535 |g 5 | ||
| matchkey_str |
article:14374331:2010----::oprsnfifrndpeintaeisoipoigeouin |
|---|---|
| hierarchy_sort_str |
2010 |
| publishDate |
2010 |
| allfields |
10.1515/CCLM.2010.109 doi artikel_Grundlieferung.pp (DE-627)NLEJ246730161 DE-627 ger DE-627 rakwb Comparison of different depletion strategies for improving resolution of the human urine proteome Walter de Gruyter 2010 5 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. Walter de Gruyter Online Zeitschriften biomarkers proteins depletion proteomics urine Magagnotti, Cinzia oth Fermo, Isabella oth Carletti, Rose Mary oth Ferrari, Maurizio oth Bachi, Angela oth Enthalten in Clinical chemistry and laboratory medicine Berlin [u.a.] : De Gruyter, 1998 48(2010), 4 vom: 12. Feb., Seite 531-535 (DE-627)NLEJ248235222 (DE-600)1492732-9 1437-4331 nnns volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 https://doi.org/10.1515/CCLM.2010.109 Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-DGR GBV_NL_ARTICLE AR 48 2010 4 12 02 531-535 5 |
| spelling |
10.1515/CCLM.2010.109 doi artikel_Grundlieferung.pp (DE-627)NLEJ246730161 DE-627 ger DE-627 rakwb Comparison of different depletion strategies for improving resolution of the human urine proteome Walter de Gruyter 2010 5 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. Walter de Gruyter Online Zeitschriften biomarkers proteins depletion proteomics urine Magagnotti, Cinzia oth Fermo, Isabella oth Carletti, Rose Mary oth Ferrari, Maurizio oth Bachi, Angela oth Enthalten in Clinical chemistry and laboratory medicine Berlin [u.a.] : De Gruyter, 1998 48(2010), 4 vom: 12. Feb., Seite 531-535 (DE-627)NLEJ248235222 (DE-600)1492732-9 1437-4331 nnns volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 https://doi.org/10.1515/CCLM.2010.109 Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-DGR GBV_NL_ARTICLE AR 48 2010 4 12 02 531-535 5 |
| allfields_unstemmed |
10.1515/CCLM.2010.109 doi artikel_Grundlieferung.pp (DE-627)NLEJ246730161 DE-627 ger DE-627 rakwb Comparison of different depletion strategies for improving resolution of the human urine proteome Walter de Gruyter 2010 5 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. Walter de Gruyter Online Zeitschriften biomarkers proteins depletion proteomics urine Magagnotti, Cinzia oth Fermo, Isabella oth Carletti, Rose Mary oth Ferrari, Maurizio oth Bachi, Angela oth Enthalten in Clinical chemistry and laboratory medicine Berlin [u.a.] : De Gruyter, 1998 48(2010), 4 vom: 12. Feb., Seite 531-535 (DE-627)NLEJ248235222 (DE-600)1492732-9 1437-4331 nnns volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 https://doi.org/10.1515/CCLM.2010.109 Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-DGR GBV_NL_ARTICLE AR 48 2010 4 12 02 531-535 5 |
| allfieldsGer |
10.1515/CCLM.2010.109 doi artikel_Grundlieferung.pp (DE-627)NLEJ246730161 DE-627 ger DE-627 rakwb Comparison of different depletion strategies for improving resolution of the human urine proteome Walter de Gruyter 2010 5 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. Walter de Gruyter Online Zeitschriften biomarkers proteins depletion proteomics urine Magagnotti, Cinzia oth Fermo, Isabella oth Carletti, Rose Mary oth Ferrari, Maurizio oth Bachi, Angela oth Enthalten in Clinical chemistry and laboratory medicine Berlin [u.a.] : De Gruyter, 1998 48(2010), 4 vom: 12. Feb., Seite 531-535 (DE-627)NLEJ248235222 (DE-600)1492732-9 1437-4331 nnns volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 https://doi.org/10.1515/CCLM.2010.109 Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-DGR GBV_NL_ARTICLE AR 48 2010 4 12 02 531-535 5 |
| allfieldsSound |
10.1515/CCLM.2010.109 doi artikel_Grundlieferung.pp (DE-627)NLEJ246730161 DE-627 ger DE-627 rakwb Comparison of different depletion strategies for improving resolution of the human urine proteome Walter de Gruyter 2010 5 Text txt rdacontent Computermedien c rdamedia Online-Ressource cr rdacarrier Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. Walter de Gruyter Online Zeitschriften biomarkers proteins depletion proteomics urine Magagnotti, Cinzia oth Fermo, Isabella oth Carletti, Rose Mary oth Ferrari, Maurizio oth Bachi, Angela oth Enthalten in Clinical chemistry and laboratory medicine Berlin [u.a.] : De Gruyter, 1998 48(2010), 4 vom: 12. Feb., Seite 531-535 (DE-627)NLEJ248235222 (DE-600)1492732-9 1437-4331 nnns volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 https://doi.org/10.1515/CCLM.2010.109 Deutschlandweit zugänglich GBV_USEFLAG_U ZDB-1-DGR GBV_NL_ARTICLE AR 48 2010 4 12 02 531-535 5 |
| source |
Enthalten in Clinical chemistry and laboratory medicine 48(2010), 4 vom: 12. Feb., Seite 531-535 volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 |
| sourceStr |
Enthalten in Clinical chemistry and laboratory medicine 48(2010), 4 vom: 12. Feb., Seite 531-535 volume:48 year:2010 number:4 day:12 month:02 pages:531-535 extent:5 |
| format_phy_str_mv |
Article |
| institution |
findex.gbv.de |
| topic_facet |
biomarkers proteins depletion proteomics urine |
| isfreeaccess_bool |
false |
| container_title |
Clinical chemistry and laboratory medicine |
| authorswithroles_txt_mv |
Magagnotti, Cinzia @@oth@@ Fermo, Isabella @@oth@@ Carletti, Rose Mary @@oth@@ Ferrari, Maurizio @@oth@@ Bachi, Angela @@oth@@ |
| publishDateDaySort_date |
2010-02-12T00:00:00Z |
| hierarchy_top_id |
NLEJ248235222 |
| id |
NLEJ246730161 |
| fullrecord |
<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ246730161</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230506145551.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">220814s2010 xx |||||o 00| ||und c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1515/CCLM.2010.109</subfield><subfield code="2">doi</subfield></datafield><datafield tag="028" ind1="5" ind2="2"><subfield code="a">artikel_Grundlieferung.pp</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ246730161</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Comparison of different depletion strategies for improving resolution of the human urine proteome</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="b">Walter de Gruyter</subfield><subfield code="c">2010</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">5</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">Text</subfield><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">Computermedien</subfield><subfield code="b">c</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield><subfield code="b">cr</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="f">Walter de Gruyter Online Zeitschriften</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">biomarkers</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">proteins depletion</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">proteomics</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">urine</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Magagnotti, Cinzia</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Fermo, Isabella</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Carletti, Rose Mary</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Ferrari, Maurizio</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Bachi, Angela</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">Enthalten in</subfield><subfield code="t">Clinical chemistry and laboratory medicine</subfield><subfield code="d">Berlin [u.a.] : De Gruyter, 1998</subfield><subfield code="g">48(2010), 4 vom: 12. Feb., Seite 531-535</subfield><subfield code="w">(DE-627)NLEJ248235222</subfield><subfield code="w">(DE-600)1492732-9</subfield><subfield code="x">1437-4331</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:48</subfield><subfield code="g">year:2010</subfield><subfield code="g">number:4</subfield><subfield code="g">day:12</subfield><subfield code="g">month:02</subfield><subfield code="g">pages:531-535</subfield><subfield code="g">extent:5</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">https://doi.org/10.1515/CCLM.2010.109</subfield><subfield code="z">Deutschlandweit zugänglich</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-DGR</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">48</subfield><subfield code="j">2010</subfield><subfield code="e">4</subfield><subfield code="b">12</subfield><subfield code="c">02</subfield><subfield code="h">531-535</subfield><subfield code="g">5</subfield></datafield></record></collection>
|
| series2 |
Walter de Gruyter Online Zeitschriften |
| ppnlink_with_tag_str_mv |
@@773@@(DE-627)NLEJ248235222 |
| format |
electronic Article |
| delete_txt_mv |
keep |
| collection |
NL |
| remote_str |
true |
| illustrated |
Not Illustrated |
| issn |
1437-4331 |
| topic_title |
Comparison of different depletion strategies for improving resolution of the human urine proteome biomarkers proteins depletion proteomics urine |
| publisher |
Walter de Gruyter |
| publisherStr |
Walter de Gruyter |
| topic |
misc biomarkers misc proteins depletion misc proteomics misc urine |
| spellingShingle |
misc biomarkers misc proteins depletion misc proteomics misc urine Comparison of different depletion strategies for improving resolution of the human urine proteome |
| topic_unstemmed |
misc biomarkers misc proteins depletion misc proteomics misc urine |
| topic_browse |
misc biomarkers misc proteins depletion misc proteomics misc urine |
| format_facet |
Elektronische Aufsätze Aufsätze Elektronische Ressource |
| format_main_str_mv |
Text Zeitschrift/Artikel |
| carriertype_str_mv |
cr |
| author2_variant |
c m cm i f if r m c rm rmc m f mf a b ab |
| hierarchy_parent_title |
Clinical chemistry and laboratory medicine |
| hierarchy_parent_id |
NLEJ248235222 |
| hierarchy_top_title |
Clinical chemistry and laboratory medicine |
| isfreeaccess_txt |
false |
| familylinks_str_mv |
(DE-627)NLEJ248235222 (DE-600)1492732-9 |
| title |
Comparison of different depletion strategies for improving resolution of the human urine proteome |
| ctrlnum |
(DE-627)NLEJ246730161 |
| title_full |
Comparison of different depletion strategies for improving resolution of the human urine proteome |
| journal |
Clinical chemistry and laboratory medicine |
| journalStr |
Clinical chemistry and laboratory medicine |
| isOA_bool |
false |
| recordtype |
marc |
| publishDateSort |
2010 |
| contenttype_str_mv |
txt |
| container_start_page |
531 |
| container_volume |
48 |
| physical |
5 |
| format_se |
Elektronische Aufsätze |
| doi_str_mv |
10.1515/CCLM.2010.109 |
| title_sort |
comparison of different depletion strategies for improving resolution of the human urine proteome |
| title_auth |
Comparison of different depletion strategies for improving resolution of the human urine proteome |
| abstract |
Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. |
| abstractGer |
Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. |
| abstract_unstemmed |
Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5. |
| collection_details |
GBV_USEFLAG_U ZDB-1-DGR GBV_NL_ARTICLE |
| container_issue |
4 |
| title_short |
Comparison of different depletion strategies for improving resolution of the human urine proteome |
| url |
https://doi.org/10.1515/CCLM.2010.109 |
| remote_bool |
true |
| author2 |
Magagnotti, Cinzia Fermo, Isabella Carletti, Rose Mary Ferrari, Maurizio Bachi, Angela |
| author2Str |
Magagnotti, Cinzia Fermo, Isabella Carletti, Rose Mary Ferrari, Maurizio Bachi, Angela |
| ppnlink |
NLEJ248235222 |
| mediatype_str_mv |
c |
| isOA_txt |
false |
| hochschulschrift_bool |
false |
| author2_role |
oth oth oth oth oth |
| doi_str |
10.1515/CCLM.2010.109 |
| up_date |
2024-07-06T09:12:44.226Z |
| _version_ |
1803820376236490752 |
| fullrecord_marcxml |
<?xml version="1.0" encoding="UTF-8"?><collection xmlns="http://www.loc.gov/MARC21/slim"><record><leader>01000caa a22002652 4500</leader><controlfield tag="001">NLEJ246730161</controlfield><controlfield tag="003">DE-627</controlfield><controlfield tag="005">20230506145551.0</controlfield><controlfield tag="007">cr uuu---uuuuu</controlfield><controlfield tag="008">220814s2010 xx |||||o 00| ||und c</controlfield><datafield tag="024" ind1="7" ind2=" "><subfield code="a">10.1515/CCLM.2010.109</subfield><subfield code="2">doi</subfield></datafield><datafield tag="028" ind1="5" ind2="2"><subfield code="a">artikel_Grundlieferung.pp</subfield></datafield><datafield tag="035" ind1=" " ind2=" "><subfield code="a">(DE-627)NLEJ246730161</subfield></datafield><datafield tag="040" ind1=" " ind2=" "><subfield code="a">DE-627</subfield><subfield code="b">ger</subfield><subfield code="c">DE-627</subfield><subfield code="e">rakwb</subfield></datafield><datafield tag="245" ind1="1" ind2="0"><subfield code="a">Comparison of different depletion strategies for improving resolution of the human urine proteome</subfield></datafield><datafield tag="264" ind1=" " ind2="1"><subfield code="b">Walter de Gruyter</subfield><subfield code="c">2010</subfield></datafield><datafield tag="300" ind1=" " ind2=" "><subfield code="a">5</subfield></datafield><datafield tag="336" ind1=" " ind2=" "><subfield code="a">Text</subfield><subfield code="b">txt</subfield><subfield code="2">rdacontent</subfield></datafield><datafield tag="337" ind1=" " ind2=" "><subfield code="a">Computermedien</subfield><subfield code="b">c</subfield><subfield code="2">rdamedia</subfield></datafield><datafield tag="338" ind1=" " ind2=" "><subfield code="a">Online-Ressource</subfield><subfield code="b">cr</subfield><subfield code="2">rdacarrier</subfield></datafield><datafield tag="520" ind1=" " ind2=" "><subfield code="a">Background: Urine, being an ultrafiltrate of plasma, is a rich source for biomarker discovery. Since potential new disease markers are often present in low concentrations, a prefractionation/enrichment step could be useful in the discovery process. To enhance the detection of low-abundance proteins, three immuno-affinity depletion approaches were evaluated. Methods: To remove the most abundant proteins from a human urine sample, GenWay™ Spin IgY-12 kit, HPLC Agilent Hu-PL7 and a home-made column vs. human serum albumin [immuno-affinity column (IAC)] were compared. Quantification of total proteins, 2-D gel electrophoresis (2-DE), Progenesis gel images analysis and mass spectrometric proteins identification were applied to evaluate these strategies. Results: Reproducibility of depletion columns, by estimating protein content of unbound fractions, were: 343±20.0 μg, 5.8%; 186.3±13.3 μg, 7.2%; 292±20.6 μg, 8.8% [mean± standard deviation (SD), CV%], for GenWay™, Agilent and IAC methods, respectively. To isolate urinary protein after depletion, ethanol precipitation provided the highest recovery (80%). Applying 2-DE and Progenesis analysis, the number of spots visualized on the gels was 468±21, 331±7, 368±22 and 304±7 (mean±SD) for GenWay™, Agilent, IAC, and the undepleted urine pool sample, respectively, with a significant difference p<0.001 compared to the GenWay procedure. Conclusions: The sequential procedure of urine samples using multi-protein immuno-affinity depletion represents a valid tool for simplifying 2-DE analysis of the urine proteome. Particularly, the GenWay™ kit followed by ethanol precipitation was found to be the most efficient method for exploring the urine proteome. Clin Chem Lab Med 2010;48:531–5.</subfield></datafield><datafield tag="533" ind1=" " ind2=" "><subfield code="f">Walter de Gruyter Online Zeitschriften</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">biomarkers</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">proteins depletion</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">proteomics</subfield></datafield><datafield tag="650" ind1=" " ind2="4"><subfield code="a">urine</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Magagnotti, Cinzia</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Fermo, Isabella</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Carletti, Rose Mary</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Ferrari, Maurizio</subfield><subfield code="4">oth</subfield></datafield><datafield tag="700" ind1="1" ind2=" "><subfield code="a">Bachi, Angela</subfield><subfield code="4">oth</subfield></datafield><datafield tag="773" ind1="0" ind2="8"><subfield code="i">Enthalten in</subfield><subfield code="t">Clinical chemistry and laboratory medicine</subfield><subfield code="d">Berlin [u.a.] : De Gruyter, 1998</subfield><subfield code="g">48(2010), 4 vom: 12. Feb., Seite 531-535</subfield><subfield code="w">(DE-627)NLEJ248235222</subfield><subfield code="w">(DE-600)1492732-9</subfield><subfield code="x">1437-4331</subfield><subfield code="7">nnns</subfield></datafield><datafield tag="773" ind1="1" ind2="8"><subfield code="g">volume:48</subfield><subfield code="g">year:2010</subfield><subfield code="g">number:4</subfield><subfield code="g">day:12</subfield><subfield code="g">month:02</subfield><subfield code="g">pages:531-535</subfield><subfield code="g">extent:5</subfield></datafield><datafield tag="856" ind1="4" ind2="0"><subfield code="u">https://doi.org/10.1515/CCLM.2010.109</subfield><subfield code="z">Deutschlandweit zugänglich</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_USEFLAG_U</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">ZDB-1-DGR</subfield></datafield><datafield tag="912" ind1=" " ind2=" "><subfield code="a">GBV_NL_ARTICLE</subfield></datafield><datafield tag="951" ind1=" " ind2=" "><subfield code="a">AR</subfield></datafield><datafield tag="952" ind1=" " ind2=" "><subfield code="d">48</subfield><subfield code="j">2010</subfield><subfield code="e">4</subfield><subfield code="b">12</subfield><subfield code="c">02</subfield><subfield code="h">531-535</subfield><subfield code="g">5</subfield></datafield></record></collection>
|
| score |
7.4010954 |