Comparative Glycomic Mapping through Quantitative Permethylation and Stable-Isotope Labeling
Comparative glycan quantification has thus far been a challenging task due to the lack of sensitive and reproducible analytical techniques. We introduce here a combination of quantitative permethylation and isotope labeling of glycans as an approach (C-GlycoMAP) allowing precise comparison between d...
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| Veröffentlicht in: | Analytical chemistry (Washington) 2007-08, Vol.79 (16), p.6064-6073 |
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| creator | Kang, Pilsoo Mechref, Yehia Kyselova, Zuzana Goetz, John A Novotny, Milos V |
| description | Comparative glycan quantification has thus far been a challenging task due to the lack of sensitive and reproducible analytical techniques. We introduce here a combination of quantitative permethylation and isotope labeling of glycans as an approach (C-GlycoMAP) allowing precise comparison between different samples in a single MALDI-MS analysis. Samples are either methylated or deuteriomethylated prior to their mixing and mass spectrometric acquisitions. Comparative analyses are based on the ratio of the two isotopically distinct forms of the same glycan structure, thus allowing a direct absolute evaluation of the intensities of the two forms originating from two different biological samples (e.g., control and diseased). The direct comparison between the two forms eliminates a MALDI-MS low m/z bias commonly associated with this technique. These comparative analyses are highly reliable when the intensity ratios of the two forms lie between 0.125 and 6, an overall reproducibility better than 30% (RSD). The value of C-GlycoMAP is demonstrated here for N-glycans derived from human blood serum collected from a healthy individual and a breast cancer patient as well as for O-glycans derived from normal and cancer cell extracts. |
| doi_str_mv | 10.1021/ac062098r |
| format | Article |
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We introduce here a combination of quantitative permethylation and isotope labeling of glycans as an approach (C-GlycoMAP) allowing precise comparison between different samples in a single MALDI-MS analysis. Samples are either methylated or deuteriomethylated prior to their mixing and mass spectrometric acquisitions. Comparative analyses are based on the ratio of the two isotopically distinct forms of the same glycan structure, thus allowing a direct absolute evaluation of the intensities of the two forms originating from two different biological samples (e.g., control and diseased). The direct comparison between the two forms eliminates a MALDI-MS low m/z bias commonly associated with this technique. These comparative analyses are highly reliable when the intensity ratios of the two forms lie between 0.125 and 6, an overall reproducibility better than 30% (RSD). 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Chem</addtitle><description>Comparative glycan quantification has thus far been a challenging task due to the lack of sensitive and reproducible analytical techniques. We introduce here a combination of quantitative permethylation and isotope labeling of glycans as an approach (C-GlycoMAP) allowing precise comparison between different samples in a single MALDI-MS analysis. Samples are either methylated or deuteriomethylated prior to their mixing and mass spectrometric acquisitions. Comparative analyses are based on the ratio of the two isotopically distinct forms of the same glycan structure, thus allowing a direct absolute evaluation of the intensities of the two forms originating from two different biological samples (e.g., control and diseased). The direct comparison between the two forms eliminates a MALDI-MS low m/z bias commonly associated with this technique. These comparative analyses are highly reliable when the intensity ratios of the two forms lie between 0.125 and 6, an overall reproducibility better than 30% (RSD). 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Mechref, Yehia ; Kyselova, Zuzana ; Goetz, John A ; Novotny, Milos V</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-a505t-7873f854fc7d833de5ccce92398d00dfb9cd9b424af8fad1406cb2a396e031763</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Analytical chemistry</topic><topic>Body fluids</topic><topic>Breast Neoplasms - blood</topic><topic>Chemistry</topic><topic>Comparative analysis</topic><topic>Exact sciences and technology</topic><topic>Female</topic><topic>Humans</topic><topic>Isotope Labeling</topic><topic>Isotopes</topic><topic>Methylation</topic><topic>Polysaccharides - blood</topic><topic>Proteins</topic><topic>Spectrometric and optical methods</topic><topic>Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kang, Pilsoo</creatorcontrib><creatorcontrib>Mechref, Yehia</creatorcontrib><creatorcontrib>Kyselova, Zuzana</creatorcontrib><creatorcontrib>Goetz, John A</creatorcontrib><creatorcontrib>Novotny, Milos V</creatorcontrib><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Aluminium Industry Abstracts</collection><collection>Biotechnology Research Abstracts</collection><collection>Ceramic Abstracts</collection><collection>Computer and Information Systems Abstracts</collection><collection>Corrosion Abstracts</collection><collection>Electronics & Communications Abstracts</collection><collection>Engineered Materials Abstracts</collection><collection>Materials Business File</collection><collection>Mechanical & Transportation Engineering Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Solid State and Superconductivity Abstracts</collection><collection>Toxicology Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>METADEX</collection><collection>Technology Research Database</collection><collection>Environmental Sciences and Pollution Management</collection><collection>ANTE: Abstracts in New Technology & Engineering</collection><collection>Engineering Research Database</collection><collection>Aerospace Database</collection><collection>Copper Technical Reference Library</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Materials Research Database</collection><collection>ProQuest Computer Science Collection</collection><collection>Civil Engineering Abstracts</collection><collection>Advanced Technologies Database with Aerospace</collection><collection>Computer and Information Systems Abstracts Academic</collection><collection>Computer and Information Systems Abstracts Professional</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical chemistry (Washington)</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kang, Pilsoo</au><au>Mechref, Yehia</au><au>Kyselova, Zuzana</au><au>Goetz, John A</au><au>Novotny, Milos V</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Comparative Glycomic Mapping through Quantitative Permethylation and Stable-Isotope Labeling</atitle><jtitle>Analytical chemistry (Washington)</jtitle><addtitle>Anal. 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| subjects | Analytical chemistry Body fluids Breast Neoplasms - blood Chemistry Comparative analysis Exact sciences and technology Female Humans Isotope Labeling Isotopes Methylation Polysaccharides - blood Proteins Spectrometric and optical methods Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization - methods |
| title | Comparative Glycomic Mapping through Quantitative Permethylation and Stable-Isotope Labeling |
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