Multidimensional liquid chromatography platform for profiling alterations of clusterin N-glycosylation in the plasma of patients with renal cell carcinoma
► Multidimensional HPLC platform for high efficiency enrichment of clusterin from plasma. ► Combined with 2-DE, high purity clusterin in μg quantity needed for glycomic analysis was obtained. ► Application of the platform to N-glycomic characterization of enriched clusterin described. ► Alterations...
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| Veröffentlicht in: | Journal of Chromatography A 2012-09, Vol.1256, p.121-128 |
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| creator | Tousi, Fateme Bones, Jonathan Iliopoulos, Othon Hancock, William S. Hincapie, Marina |
| description | ► Multidimensional HPLC platform for high efficiency enrichment of clusterin from plasma. ► Combined with 2-DE, high purity clusterin in μg quantity needed for glycomic analysis was obtained. ► Application of the platform to N-glycomic characterization of enriched clusterin described. ► Alterations in clusterin glycosylation associated with the presence of the cancer were observed. ► Observed alterations in glycosylation may serve as potential markers with further verification.
Identification of potential changes in the glycosylation of existing cancer biomarkers can result in a higher level of diagnostic sensitivity and specificity. Clusterin (Apolipoprotein J) has been implicated in renal cell carcinoma (RCC) and other types of malignancy as potential biomarker. In the present work, an automated multi-dimensional HPLC platform enabling high throughput affinity enrichment of clusterin from plasma samples was developed. Integrated with two dimensional gel electrophoresis, high purity clusterin in microgram quantities suitable for glycan characterization was isolated. The analytical platform was applied to study clusterin glycosylation in a small group of RCC patients before and after nephrectopy as a pilot study to evaluate the performance of the platform. A statistically significant decrease was observed in the levels of a bi-antennary digalactosyl disialylated (A2G2S(3)2) glycans while the levels of a core fucosylated bi-antennary digalactosyl disialylated glycan (FA2G2S(6)2) and a tri-antennary trigalactosyl disialylated glycan (A3G3S(6)2) were increased in the post-surgery plasma samples. |
| doi_str_mv | 10.1016/j.chroma.2012.07.066 |
| format | Article |
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Identification of potential changes in the glycosylation of existing cancer biomarkers can result in a higher level of diagnostic sensitivity and specificity. Clusterin (Apolipoprotein J) has been implicated in renal cell carcinoma (RCC) and other types of malignancy as potential biomarker. In the present work, an automated multi-dimensional HPLC platform enabling high throughput affinity enrichment of clusterin from plasma samples was developed. Integrated with two dimensional gel electrophoresis, high purity clusterin in microgram quantities suitable for glycan characterization was isolated. The analytical platform was applied to study clusterin glycosylation in a small group of RCC patients before and after nephrectopy as a pilot study to evaluate the performance of the platform. A statistically significant decrease was observed in the levels of a bi-antennary digalactosyl disialylated (A2G2S(3)2) glycans while the levels of a core fucosylated bi-antennary digalactosyl disialylated glycan (FA2G2S(6)2) and a tri-antennary trigalactosyl disialylated glycan (A3G3S(6)2) were increased in the post-surgery plasma samples.</description><identifier>ISSN: 0021-9673</identifier><identifier>EISSN: 1873-3778</identifier><identifier>DOI: 10.1016/j.chroma.2012.07.066</identifier><identifier>PMID: 22885037</identifier><language>eng</language><publisher>Netherlands: Elsevier B.V</publisher><subject>biomarkers ; carcinoma ; Carcinoma, Renal Cell - blood ; Carcinoma, Renal Cell - metabolism ; Chromatography, Liquid - methods ; Clusterin ; Clusterin - blood ; Clusterin - metabolism ; diagnostic sensitivity ; Electrophoresis, Gel, Two-Dimensional ; Glycosylation ; high performance liquid chromatography ; Humans ; Kidney Neoplasms - blood ; Kidney Neoplasms - metabolism ; Multi-dimensional HPLC ; N-glycan ; patients ; polysaccharides ; Renal cell carcinoma ; Two dimensional gel electrophoresis (2-DE) ; two-dimensional gel electrophoresis ; UPLC–HILIC</subject><ispartof>Journal of Chromatography A, 2012-09, Vol.1256, p.121-128</ispartof><rights>2012 Elsevier B.V.</rights><rights>Copyright © 2012 Elsevier B.V. All rights reserved.</rights><rights>2012 Elsevier B.V. All rights reserved. 2012</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c553t-7aca39819c06096518d42effba97b03d44dd9f86ac717856c8fad27fe643e11f3</citedby><cites>FETCH-LOGICAL-c553t-7aca39819c06096518d42effba97b03d44dd9f86ac717856c8fad27fe643e11f3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S0021967312011363$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>230,314,776,780,881,3536,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22885037$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Tousi, Fateme</creatorcontrib><creatorcontrib>Bones, Jonathan</creatorcontrib><creatorcontrib>Iliopoulos, Othon</creatorcontrib><creatorcontrib>Hancock, William S.</creatorcontrib><creatorcontrib>Hincapie, Marina</creatorcontrib><title>Multidimensional liquid chromatography platform for profiling alterations of clusterin N-glycosylation in the plasma of patients with renal cell carcinoma</title><title>Journal of Chromatography A</title><addtitle>J Chromatogr A</addtitle><description>► Multidimensional HPLC platform for high efficiency enrichment of clusterin from plasma. ► Combined with 2-DE, high purity clusterin in μg quantity needed for glycomic analysis was obtained. ► Application of the platform to N-glycomic characterization of enriched clusterin described. ► Alterations in clusterin glycosylation associated with the presence of the cancer were observed. ► Observed alterations in glycosylation may serve as potential markers with further verification.
Identification of potential changes in the glycosylation of existing cancer biomarkers can result in a higher level of diagnostic sensitivity and specificity. Clusterin (Apolipoprotein J) has been implicated in renal cell carcinoma (RCC) and other types of malignancy as potential biomarker. In the present work, an automated multi-dimensional HPLC platform enabling high throughput affinity enrichment of clusterin from plasma samples was developed. Integrated with two dimensional gel electrophoresis, high purity clusterin in microgram quantities suitable for glycan characterization was isolated. The analytical platform was applied to study clusterin glycosylation in a small group of RCC patients before and after nephrectopy as a pilot study to evaluate the performance of the platform. A statistically significant decrease was observed in the levels of a bi-antennary digalactosyl disialylated (A2G2S(3)2) glycans while the levels of a core fucosylated bi-antennary digalactosyl disialylated glycan (FA2G2S(6)2) and a tri-antennary trigalactosyl disialylated glycan (A3G3S(6)2) were increased in the post-surgery plasma samples.</description><subject>biomarkers</subject><subject>carcinoma</subject><subject>Carcinoma, Renal Cell - blood</subject><subject>Carcinoma, Renal Cell - metabolism</subject><subject>Chromatography, Liquid - methods</subject><subject>Clusterin</subject><subject>Clusterin - blood</subject><subject>Clusterin - metabolism</subject><subject>diagnostic sensitivity</subject><subject>Electrophoresis, Gel, Two-Dimensional</subject><subject>Glycosylation</subject><subject>high performance liquid chromatography</subject><subject>Humans</subject><subject>Kidney Neoplasms - blood</subject><subject>Kidney Neoplasms - metabolism</subject><subject>Multi-dimensional HPLC</subject><subject>N-glycan</subject><subject>patients</subject><subject>polysaccharides</subject><subject>Renal cell carcinoma</subject><subject>Two dimensional gel electrophoresis (2-DE)</subject><subject>two-dimensional gel electrophoresis</subject><subject>UPLC–HILIC</subject><issn>0021-9673</issn><issn>1873-3778</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFUsFu1DAQtRCIbhf-AIGPXBLsOLGdCxKqoCAVOEDPltexE6-cOLWdov0VvhaHlAIXuNjSzJs3b-YNAM8wKjHC9NWxVEPwoywrhKsSsRJR-gDsMGekIIzxh2CHUIWLljJyBs5jPCKEGWLVY3BWVZw3iLAd-P5xccl2dtRTtH6SDjp7s9gObuTJ90HOwwnOTibjwwjzA-fgjXV26qF0SQeZcmWE3kDllpgDdoKfit6dlI8n9zMLcygNeqWJo1yhc47rKUX4zaYBBr22VtrlRwZlp9z7CXhkpIv66d2_B9fv3n69eF9cfb78cPHmqlBNQ1LBpJKk5bhViKKWNph3daWNOciWHRDp6rrrWsOpVAwz3lDFjewqZjSticbYkD14vfHOy2HUncqqgnRiDnaU4SS8tOLvzGQH0ftbUZO2yiSZ4OUdQfA3i45JjDaus8hJ-yUKTNu6ZpRkuf-HkgZhvpq4B_UGVcHHGLS5V4SRWC9AHMVmklgvQCAm8gXksud_TnNf9MvyDHixAYz0QvbBRnH9JTM0-Tx427Dm90J03vqt1UFElb1SurNBqyQ6b_-t4QfdqNMJ</recordid><startdate>20120921</startdate><enddate>20120921</enddate><creator>Tousi, Fateme</creator><creator>Bones, Jonathan</creator><creator>Iliopoulos, Othon</creator><creator>Hancock, William S.</creator><creator>Hincapie, Marina</creator><general>Elsevier B.V</general><scope>FBQ</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QH</scope><scope>7UA</scope><scope>C1K</scope><scope>F1W</scope><scope>H97</scope><scope>L.G</scope><scope>7S9</scope><scope>L.6</scope><scope>5PM</scope></search><sort><creationdate>20120921</creationdate><title>Multidimensional liquid chromatography platform for profiling alterations of clusterin N-glycosylation in the plasma of patients with renal cell carcinoma</title><author>Tousi, Fateme ; Bones, Jonathan ; Iliopoulos, Othon ; Hancock, William S. ; Hincapie, Marina</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c553t-7aca39819c06096518d42effba97b03d44dd9f86ac717856c8fad27fe643e11f3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>biomarkers</topic><topic>carcinoma</topic><topic>Carcinoma, Renal Cell - blood</topic><topic>Carcinoma, Renal Cell - metabolism</topic><topic>Chromatography, Liquid - methods</topic><topic>Clusterin</topic><topic>Clusterin - blood</topic><topic>Clusterin - metabolism</topic><topic>diagnostic sensitivity</topic><topic>Electrophoresis, Gel, Two-Dimensional</topic><topic>Glycosylation</topic><topic>high performance liquid chromatography</topic><topic>Humans</topic><topic>Kidney Neoplasms - blood</topic><topic>Kidney Neoplasms - metabolism</topic><topic>Multi-dimensional HPLC</topic><topic>N-glycan</topic><topic>patients</topic><topic>polysaccharides</topic><topic>Renal cell carcinoma</topic><topic>Two dimensional gel electrophoresis (2-DE)</topic><topic>two-dimensional gel electrophoresis</topic><topic>UPLC–HILIC</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Tousi, Fateme</creatorcontrib><creatorcontrib>Bones, Jonathan</creatorcontrib><creatorcontrib>Iliopoulos, Othon</creatorcontrib><creatorcontrib>Hancock, William S.</creatorcontrib><creatorcontrib>Hincapie, Marina</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aqualine</collection><collection>Water Resources Abstracts</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ASFA: Aquatic Sciences and Fisheries Abstracts</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) 3: Aquatic Pollution & Environmental Quality</collection><collection>Aquatic Science & Fisheries Abstracts (ASFA) Professional</collection><collection>AGRICOLA</collection><collection>AGRICOLA - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Journal of Chromatography A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Tousi, Fateme</au><au>Bones, Jonathan</au><au>Iliopoulos, Othon</au><au>Hancock, William S.</au><au>Hincapie, Marina</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Multidimensional liquid chromatography platform for profiling alterations of clusterin N-glycosylation in the plasma of patients with renal cell carcinoma</atitle><jtitle>Journal of Chromatography A</jtitle><addtitle>J Chromatogr A</addtitle><date>2012-09-21</date><risdate>2012</risdate><volume>1256</volume><spage>121</spage><epage>128</epage><pages>121-128</pages><issn>0021-9673</issn><eissn>1873-3778</eissn><abstract>► Multidimensional HPLC platform for high efficiency enrichment of clusterin from plasma. ► Combined with 2-DE, high purity clusterin in μg quantity needed for glycomic analysis was obtained. ► Application of the platform to N-glycomic characterization of enriched clusterin described. ► Alterations in clusterin glycosylation associated with the presence of the cancer were observed. ► Observed alterations in glycosylation may serve as potential markers with further verification.
Identification of potential changes in the glycosylation of existing cancer biomarkers can result in a higher level of diagnostic sensitivity and specificity. Clusterin (Apolipoprotein J) has been implicated in renal cell carcinoma (RCC) and other types of malignancy as potential biomarker. In the present work, an automated multi-dimensional HPLC platform enabling high throughput affinity enrichment of clusterin from plasma samples was developed. Integrated with two dimensional gel electrophoresis, high purity clusterin in microgram quantities suitable for glycan characterization was isolated. The analytical platform was applied to study clusterin glycosylation in a small group of RCC patients before and after nephrectopy as a pilot study to evaluate the performance of the platform. A statistically significant decrease was observed in the levels of a bi-antennary digalactosyl disialylated (A2G2S(3)2) glycans while the levels of a core fucosylated bi-antennary digalactosyl disialylated glycan (FA2G2S(6)2) and a tri-antennary trigalactosyl disialylated glycan (A3G3S(6)2) were increased in the post-surgery plasma samples.</abstract><cop>Netherlands</cop><pub>Elsevier B.V</pub><pmid>22885037</pmid><doi>10.1016/j.chroma.2012.07.066</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | biomarkers carcinoma Carcinoma, Renal Cell - blood Carcinoma, Renal Cell - metabolism Chromatography, Liquid - methods Clusterin Clusterin - blood Clusterin - metabolism diagnostic sensitivity Electrophoresis, Gel, Two-Dimensional Glycosylation high performance liquid chromatography Humans Kidney Neoplasms - blood Kidney Neoplasms - metabolism Multi-dimensional HPLC N-glycan patients polysaccharides Renal cell carcinoma Two dimensional gel electrophoresis (2-DE) two-dimensional gel electrophoresis UPLC–HILIC |
| title | Multidimensional liquid chromatography platform for profiling alterations of clusterin N-glycosylation in the plasma of patients with renal cell carcinoma |
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