Analysis of N-glycans from recombinant immunoglobulin G by on-line reversed-phase high-performance liquid chromatography/mass spectrometry
An on-line reversed-phase (RP) high-performance liquid chromatography/mass spectrometry (MS) method has been developed for profiling and characterizing N-glycans from recombinant immunoglobulin G antibodies. In this method, released N-glycans are derivatized at their reducing end with 2-aminobenzami...
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| Veröffentlicht in: | Analytical biochemistry 2007-11, Vol.370 (2), p.147-161 |
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| container_title | Analytical biochemistry |
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| creator | Chen, Xiaoyu Flynn, Gregory C. |
| description | An on-line reversed-phase (RP) high-performance liquid chromatography/mass spectrometry (MS) method has been developed for profiling and characterizing
N-glycans from recombinant immunoglobulin G antibodies. In this method, released
N-glycans are derivatized at their reducing end with 2-aminobenzamide (2AB) and separated on a RP column with on-line fluorescence and MS detection. The method achieves good resolution of all major glycans and segregates glycan types (high-mannose, hybrid, and complex) to different regions of the chromatogram, thus allowing accurate quantification of
N-glycans from the fluorescent signal alone. Moreover, the mobile phase used allows high quality on-line MS detection. The 2AB-labeled
N-glycans demonstrate good ionization efficiency in electrospray and generate primarily doubly charged [M+2H]
2+ ions. The mass and structural information can be readily obtained from the on-line MS and tandem MS data. As little as 70 fmol glycan species can be detected and identified. |
| doi_str_mv | 10.1016/j.ab.2007.08.012 |
| format | Article |
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N-glycans from recombinant immunoglobulin G antibodies. In this method, released
N-glycans are derivatized at their reducing end with 2-aminobenzamide (2AB) and separated on a RP column with on-line fluorescence and MS detection. The method achieves good resolution of all major glycans and segregates glycan types (high-mannose, hybrid, and complex) to different regions of the chromatogram, thus allowing accurate quantification of
N-glycans from the fluorescent signal alone. Moreover, the mobile phase used allows high quality on-line MS detection. The 2AB-labeled
N-glycans demonstrate good ionization efficiency in electrospray and generate primarily doubly charged [M+2H]
2+ ions. The mass and structural information can be readily obtained from the on-line MS and tandem MS data. As little as 70 fmol glycan species can be detected and identified.</description><identifier>ISSN: 0003-2697</identifier><identifier>EISSN: 1096-0309</identifier><identifier>DOI: 10.1016/j.ab.2007.08.012</identifier><identifier>PMID: 17880905</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Antibody ; Carbohydrate Conformation ; Chromatography, High Pressure Liquid ; Immunoglobulin G - chemistry ; Indicators and Reagents ; LC/MS ; Mass Spectrometry ; Models, Molecular ; N-linked glycans ; Oligosaccharides ; Polysaccharides - chemistry ; Polysaccharides - isolation & purification ; Recombinant Proteins - chemistry ; Tandem MS</subject><ispartof>Analytical biochemistry, 2007-11, Vol.370 (2), p.147-161</ispartof><rights>2007 Elsevier Inc.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c414t-8f6fe8ba0572c0020c5ef392e4a5bc75d060303973525de233dc84558d25d5ac3</citedby><cites>FETCH-LOGICAL-c414t-8f6fe8ba0572c0020c5ef392e4a5bc75d060303973525de233dc84558d25d5ac3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.ab.2007.08.012$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,780,784,3550,27924,27925,45995</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/17880905$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Chen, Xiaoyu</creatorcontrib><creatorcontrib>Flynn, Gregory C.</creatorcontrib><title>Analysis of N-glycans from recombinant immunoglobulin G by on-line reversed-phase high-performance liquid chromatography/mass spectrometry</title><title>Analytical biochemistry</title><addtitle>Anal Biochem</addtitle><description>An on-line reversed-phase (RP) high-performance liquid chromatography/mass spectrometry (MS) method has been developed for profiling and characterizing
N-glycans from recombinant immunoglobulin G antibodies. In this method, released
N-glycans are derivatized at their reducing end with 2-aminobenzamide (2AB) and separated on a RP column with on-line fluorescence and MS detection. The method achieves good resolution of all major glycans and segregates glycan types (high-mannose, hybrid, and complex) to different regions of the chromatogram, thus allowing accurate quantification of
N-glycans from the fluorescent signal alone. Moreover, the mobile phase used allows high quality on-line MS detection. The 2AB-labeled
N-glycans demonstrate good ionization efficiency in electrospray and generate primarily doubly charged [M+2H]
2+ ions. The mass and structural information can be readily obtained from the on-line MS and tandem MS data. As little as 70 fmol glycan species can be detected and identified.</description><subject>Antibody</subject><subject>Carbohydrate Conformation</subject><subject>Chromatography, High Pressure Liquid</subject><subject>Immunoglobulin G - chemistry</subject><subject>Indicators and Reagents</subject><subject>LC/MS</subject><subject>Mass Spectrometry</subject><subject>Models, Molecular</subject><subject>N-linked glycans</subject><subject>Oligosaccharides</subject><subject>Polysaccharides - chemistry</subject><subject>Polysaccharides - isolation & purification</subject><subject>Recombinant Proteins - chemistry</subject><subject>Tandem MS</subject><issn>0003-2697</issn><issn>1096-0309</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2007</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kE1v1DAQhi0Eokvhzgn5xC3pOI4Th1tVlYJUwQXOlmNPNl7FcWonlfIX-NW42pU4cZoPPfNK8xDykUHJgDU3p1L3ZQXQliBLYNUrcmDQNQVw6F6TAwDwomq69oq8S-kEwFgtmrfkirVSQgfiQP7cznrak0s0DPRHcZx2o-dEhxg8jWiC792s55U677c5HKfQb5Ob6QPtdxrmIveYuWeMCW2xjDohHd1xLBaMQ4hezwbp5J42Z6kZc6hewzHqZdxvvE6JpgXNmte4xv09eTPoKeGHS70mv7_e_7r7Vjz-fPh-d_tYmJrVayGHZkDZaxBtZQAqMAIH3lVYa9GbVlho8vu8a7mohMWKc2tkLYS0eRTa8Gvy-Zy7xPC0YVqVd8ngNOkZw5ZUI3m2JHgG4QyaGFKKOKglOq_jrhioF__qpHSvXvwrkCr7zyefLtlb79H-O7gIz8CXM4D5w2eHUSXjMFuyLutelQ3u_-l_AQW3mBI</recordid><startdate>20071115</startdate><enddate>20071115</enddate><creator>Chen, Xiaoyu</creator><creator>Flynn, Gregory C.</creator><general>Elsevier Inc</general><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>7X8</scope></search><sort><creationdate>20071115</creationdate><title>Analysis of N-glycans from recombinant immunoglobulin G by on-line reversed-phase high-performance liquid chromatography/mass spectrometry</title><author>Chen, Xiaoyu ; Flynn, Gregory C.</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c414t-8f6fe8ba0572c0020c5ef392e4a5bc75d060303973525de233dc84558d25d5ac3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2007</creationdate><topic>Antibody</topic><topic>Carbohydrate Conformation</topic><topic>Chromatography, High Pressure Liquid</topic><topic>Immunoglobulin G - chemistry</topic><topic>Indicators and Reagents</topic><topic>LC/MS</topic><topic>Mass Spectrometry</topic><topic>Models, Molecular</topic><topic>N-linked glycans</topic><topic>Oligosaccharides</topic><topic>Polysaccharides - chemistry</topic><topic>Polysaccharides - isolation & purification</topic><topic>Recombinant Proteins - chemistry</topic><topic>Tandem MS</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Chen, Xiaoyu</creatorcontrib><creatorcontrib>Flynn, Gregory C.</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Analytical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Chen, Xiaoyu</au><au>Flynn, Gregory C.</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Analysis of N-glycans from recombinant immunoglobulin G by on-line reversed-phase high-performance liquid chromatography/mass spectrometry</atitle><jtitle>Analytical biochemistry</jtitle><addtitle>Anal Biochem</addtitle><date>2007-11-15</date><risdate>2007</risdate><volume>370</volume><issue>2</issue><spage>147</spage><epage>161</epage><pages>147-161</pages><issn>0003-2697</issn><eissn>1096-0309</eissn><abstract>An on-line reversed-phase (RP) high-performance liquid chromatography/mass spectrometry (MS) method has been developed for profiling and characterizing
N-glycans from recombinant immunoglobulin G antibodies. In this method, released
N-glycans are derivatized at their reducing end with 2-aminobenzamide (2AB) and separated on a RP column with on-line fluorescence and MS detection. The method achieves good resolution of all major glycans and segregates glycan types (high-mannose, hybrid, and complex) to different regions of the chromatogram, thus allowing accurate quantification of
N-glycans from the fluorescent signal alone. Moreover, the mobile phase used allows high quality on-line MS detection. The 2AB-labeled
N-glycans demonstrate good ionization efficiency in electrospray and generate primarily doubly charged [M+2H]
2+ ions. The mass and structural information can be readily obtained from the on-line MS and tandem MS data. As little as 70 fmol glycan species can be detected and identified.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>17880905</pmid><doi>10.1016/j.ab.2007.08.012</doi><tpages>15</tpages></addata></record> |
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| ispartof | Analytical biochemistry, 2007-11, Vol.370 (2), p.147-161 |
| issn | 0003-2697 1096-0309 |
| language | eng |
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| source | MEDLINE; Access via ScienceDirect (Elsevier) |
| subjects | Antibody Carbohydrate Conformation Chromatography, High Pressure Liquid Immunoglobulin G - chemistry Indicators and Reagents LC/MS Mass Spectrometry Models, Molecular N-linked glycans Oligosaccharides Polysaccharides - chemistry Polysaccharides - isolation & purification Recombinant Proteins - chemistry Tandem MS |
| title | Analysis of N-glycans from recombinant immunoglobulin G by on-line reversed-phase high-performance liquid chromatography/mass spectrometry |
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