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Absolute quantification of podocalyxin, a potential biomarker of glomerular injury in human urine, by liquid chromatography–mass spectrometry
•A quantification method of human podocalyxin in urine by LC–MS/MS has been developed.•Inter assay precisions and accuracies are
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| Published in: | Journal of Chromatography A 2015-06, Vol.1397, p.81-85 |
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| Main Authors: | , , , , , , , |
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| Language: | English |
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| cited_by | cdi_FETCH-LOGICAL-c466t-dd7c94f9cca1488eccb768dc98f69fbbe8eb95718bd7169ec44283706ceab06b3 |
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| cites | cdi_FETCH-LOGICAL-c466t-dd7c94f9cca1488eccb768dc98f69fbbe8eb95718bd7169ec44283706ceab06b3 |
| container_end_page | 85 |
| container_issue | |
| container_start_page | 81 |
| container_title | Journal of Chromatography A |
| container_volume | 1397 |
| creator | Biarc, Jordane Simon, Romain Fonbonne, Catherine Léonard, Jean-François Gautier, Jean-Charles Pasquier, Olivier Lemoine, Jerôme Salvador, Arnaud |
| description | •A quantification method of human podocalyxin in urine by LC–MS/MS has been developed.•Inter assay precisions and accuracies are |
| doi_str_mv | 10.1016/j.chroma.2015.04.003 |
| format | article |
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Podocalyxin is a protein present in specialized glomerulus cells called podocytes and may be released in the urine in case of kidney injury. In this context, its quantification could be of great interest in order to monitor glomerular injury. Liquid chromatography tandem mass spectrometry (LC–MS/MS), in selected reaction monitoring (SRM) mode, has been demonstrated as a powerful technique that can be applied to protein quantification. This paper describes the development of a quantification method of human podocalyxin in urine by LC–MS/MS in SRM mode by monitoring one proteotypic peptide with an isotope-dilution standardization strategy employing 13C/15N labelled peptides. Inter/intra assay precisions and accuracies of the assay were below 10% and between 90% and 106.1%, respectively. In addition, the method was linear between 0.78 and 100ng/mL and could therefore be used to quantify endogenous level of podocalyxin that was estimated between 15.2 and 44.2ng/mL in urine samples from healthy donor.</description><identifier>ISSN: 0021-9673</identifier><identifier>EISSN: 1873-3778</identifier><identifier>DOI: 10.1016/j.chroma.2015.04.003</identifier><language>eng</language><publisher>Elsevier B.V</publisher><subject>Analytical chemistry ; Biochemistry ; Biochemistry, Molecular Biology ; Biomarker ; Chemical Sciences ; Life Sciences ; Podocalyxin ; Protein ; Quantification ; SRM ; Urine</subject><ispartof>Journal of Chromatography A, 2015-06, Vol.1397, p.81-85</ispartof><rights>2015 Elsevier B.V.</rights><rights>Distributed under a Creative Commons Attribution 4.0 International License</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c466t-dd7c94f9cca1488eccb768dc98f69fbbe8eb95718bd7169ec44283706ceab06b3</citedby><cites>FETCH-LOGICAL-c466t-dd7c94f9cca1488eccb768dc98f69fbbe8eb95718bd7169ec44283706ceab06b3</cites><orcidid>0000-0002-5981-3028 ; 0000-0002-5497-6880 ; 0000-0001-5138-0822</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>230,314,780,784,885,27923,27924</link.rule.ids><backlink>$$Uhttps://hal.science/hal-01164871$$DView record in HAL$$Hfree_for_read</backlink></links><search><creatorcontrib>Biarc, Jordane</creatorcontrib><creatorcontrib>Simon, Romain</creatorcontrib><creatorcontrib>Fonbonne, Catherine</creatorcontrib><creatorcontrib>Léonard, Jean-François</creatorcontrib><creatorcontrib>Gautier, Jean-Charles</creatorcontrib><creatorcontrib>Pasquier, Olivier</creatorcontrib><creatorcontrib>Lemoine, Jerôme</creatorcontrib><creatorcontrib>Salvador, Arnaud</creatorcontrib><title>Absolute quantification of podocalyxin, a potential biomarker of glomerular injury in human urine, by liquid chromatography–mass spectrometry</title><title>Journal of Chromatography A</title><description>•A quantification method of human podocalyxin in urine by LC–MS/MS has been developed.•Inter assay precisions and accuracies are <10% and <20% respectively.•The lower limit of quantification is 0.78ng/mL from 1mL of urine.•The quantification method is compatible with endogenous level of podocalyxin.
Podocalyxin is a protein present in specialized glomerulus cells called podocytes and may be released in the urine in case of kidney injury. In this context, its quantification could be of great interest in order to monitor glomerular injury. Liquid chromatography tandem mass spectrometry (LC–MS/MS), in selected reaction monitoring (SRM) mode, has been demonstrated as a powerful technique that can be applied to protein quantification. This paper describes the development of a quantification method of human podocalyxin in urine by LC–MS/MS in SRM mode by monitoring one proteotypic peptide with an isotope-dilution standardization strategy employing 13C/15N labelled peptides. Inter/intra assay precisions and accuracies of the assay were below 10% and between 90% and 106.1%, respectively. In addition, the method was linear between 0.78 and 100ng/mL and could therefore be used to quantify endogenous level of podocalyxin that was estimated between 15.2 and 44.2ng/mL in urine samples from healthy donor.</description><subject>Analytical chemistry</subject><subject>Biochemistry</subject><subject>Biochemistry, Molecular Biology</subject><subject>Biomarker</subject><subject>Chemical Sciences</subject><subject>Life Sciences</subject><subject>Podocalyxin</subject><subject>Protein</subject><subject>Quantification</subject><subject>SRM</subject><subject>Urine</subject><issn>0021-9673</issn><issn>1873-3778</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2015</creationdate><recordtype>article</recordtype><recordid>eNp9UctOwzAQtBBIlMcfcPAViQS7cW3nglQhXlIlLnC2bGfTuqRxsRNEbvwBB_6QL8FVEEdOq92dmd3RIHRGSU4J5Zfr3K6C3-h8SugsJywnpNhDEypFkRVCyH00IWRKs5KL4hAdxbgmhAoiphP0OTfRN30H-LXXbedqZ3XnfIt9jbe-8lY3w7trL7BObQcJoRtsXDoWXiDsUMvGbyD0jQ7Ytes-DKngVb_RLe6Da-ECmwE37rV3FR7f7Pwy6O1q-P742ugYcdyC7dICujCcoINaNxFOf-sxer69ebq-zxaPdw_X80VmGeddVlXClqwurdWUSQnWGsFlZUtZ87I2BiSYciaoNJWgvATL2FQWgnAL2hBuimN0PuqudKO2wSU_g_Laqfv5Qu1mhFLOpKBvNGHZiLXBxxig_iNQonYBqLUanaldAIowlQJItKuRBsnHm4OgonXQWqhcSIZV5d3_Aj9S7Jah</recordid><startdate>20150605</startdate><enddate>20150605</enddate><creator>Biarc, Jordane</creator><creator>Simon, Romain</creator><creator>Fonbonne, Catherine</creator><creator>Léonard, Jean-François</creator><creator>Gautier, Jean-Charles</creator><creator>Pasquier, Olivier</creator><creator>Lemoine, Jerôme</creator><creator>Salvador, Arnaud</creator><general>Elsevier B.V</general><general>Elsevier</general><scope>AAYXX</scope><scope>CITATION</scope><scope>1XC</scope><orcidid>https://orcid.org/0000-0002-5981-3028</orcidid><orcidid>https://orcid.org/0000-0002-5497-6880</orcidid><orcidid>https://orcid.org/0000-0001-5138-0822</orcidid></search><sort><creationdate>20150605</creationdate><title>Absolute quantification of podocalyxin, a potential biomarker of glomerular injury in human urine, by liquid chromatography–mass spectrometry</title><author>Biarc, Jordane ; Simon, Romain ; Fonbonne, Catherine ; Léonard, Jean-François ; Gautier, Jean-Charles ; Pasquier, Olivier ; Lemoine, Jerôme ; Salvador, Arnaud</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c466t-dd7c94f9cca1488eccb768dc98f69fbbe8eb95718bd7169ec44283706ceab06b3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2015</creationdate><topic>Analytical chemistry</topic><topic>Biochemistry</topic><topic>Biochemistry, Molecular Biology</topic><topic>Biomarker</topic><topic>Chemical Sciences</topic><topic>Life Sciences</topic><topic>Podocalyxin</topic><topic>Protein</topic><topic>Quantification</topic><topic>SRM</topic><topic>Urine</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Biarc, Jordane</creatorcontrib><creatorcontrib>Simon, Romain</creatorcontrib><creatorcontrib>Fonbonne, Catherine</creatorcontrib><creatorcontrib>Léonard, Jean-François</creatorcontrib><creatorcontrib>Gautier, Jean-Charles</creatorcontrib><creatorcontrib>Pasquier, Olivier</creatorcontrib><creatorcontrib>Lemoine, Jerôme</creatorcontrib><creatorcontrib>Salvador, Arnaud</creatorcontrib><collection>CrossRef</collection><collection>Hyper Article en Ligne (HAL)</collection><jtitle>Journal of Chromatography A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Biarc, Jordane</au><au>Simon, Romain</au><au>Fonbonne, Catherine</au><au>Léonard, Jean-François</au><au>Gautier, Jean-Charles</au><au>Pasquier, Olivier</au><au>Lemoine, Jerôme</au><au>Salvador, Arnaud</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Absolute quantification of podocalyxin, a potential biomarker of glomerular injury in human urine, by liquid chromatography–mass spectrometry</atitle><jtitle>Journal of Chromatography A</jtitle><date>2015-06-05</date><risdate>2015</risdate><volume>1397</volume><spage>81</spage><epage>85</epage><pages>81-85</pages><issn>0021-9673</issn><eissn>1873-3778</eissn><abstract>•A quantification method of human podocalyxin in urine by LC–MS/MS has been developed.•Inter assay precisions and accuracies are <10% and <20% respectively.•The lower limit of quantification is 0.78ng/mL from 1mL of urine.•The quantification method is compatible with endogenous level of podocalyxin.
Podocalyxin is a protein present in specialized glomerulus cells called podocytes and may be released in the urine in case of kidney injury. In this context, its quantification could be of great interest in order to monitor glomerular injury. Liquid chromatography tandem mass spectrometry (LC–MS/MS), in selected reaction monitoring (SRM) mode, has been demonstrated as a powerful technique that can be applied to protein quantification. This paper describes the development of a quantification method of human podocalyxin in urine by LC–MS/MS in SRM mode by monitoring one proteotypic peptide with an isotope-dilution standardization strategy employing 13C/15N labelled peptides. Inter/intra assay precisions and accuracies of the assay were below 10% and between 90% and 106.1%, respectively. In addition, the method was linear between 0.78 and 100ng/mL and could therefore be used to quantify endogenous level of podocalyxin that was estimated between 15.2 and 44.2ng/mL in urine samples from healthy donor.</abstract><pub>Elsevier B.V</pub><doi>10.1016/j.chroma.2015.04.003</doi><tpages>5</tpages><orcidid>https://orcid.org/0000-0002-5981-3028</orcidid><orcidid>https://orcid.org/0000-0002-5497-6880</orcidid><orcidid>https://orcid.org/0000-0001-5138-0822</orcidid></addata></record> |
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| subjects | Analytical chemistry Biochemistry Biochemistry, Molecular Biology Biomarker Chemical Sciences Life Sciences Podocalyxin Protein Quantification SRM Urine |
| title | Absolute quantification of podocalyxin, a potential biomarker of glomerular injury in human urine, by liquid chromatography–mass spectrometry |
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