| Rapid characterization of N-linked glycans from secreted and gel-purified monoclonal antibodies using MALDI-ToF mass spectrometry. | |
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MedLine Citation:
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PMID: 20661906 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Recombinant monoclonal antibodies (MAbs) are increasingly being used for therapeutic use and correct glycosylation of these MAbs is essential for their correct function. Glycosylation profiles are host cell- and antibody class-dependent and can change over culture time and environmental conditions. Therefore, rapid monitoring of glycan addition/status is of great importance for process validity. We describe two workflows of generally applicability for glycan profiling of purified and gel-purified MAbs produced in NS0 and CHO cells, in which small-scale antibody purification and buffer exchange is combined with PNGase F glycan cleavage and graphite HyperCarb desalting. MALDI-ToF mass spectrometry is used for sensitive detection of glycan forms, with the ability to confirm glycan structures by selective ion fragmentation. Both workflows are rapid, technically simple and amenable to automation, and use in multi-well formats. |
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Authors:
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Rasmus Hansen; Alan J Dickson; Royston Goodacre; Gill M Stephens; Christopher A Sellick |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Biotechnology and bioengineering Volume: 107 ISSN: 1097-0290 ISO Abbreviation: Biotechnol. Bioeng. Publication Date: 2010 Dec |
Date Detail:
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Created Date: 2010-10-25 Completed Date: 2011-01-31 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 7502021 Medline TA: Biotechnol Bioeng Country: United States |
Other Details:
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Languages: eng Pagination: 902-8 Citation Subset: IM |
Copyright Information:
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© 2010 Wiley Periodicals, Inc. |
Affiliation:
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Faculty of Life Sciences, The University of Manchester, Michael Smith Building, Oxford Road, Manchester M13 9PT, UK. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Animals Antibodies, Monoclonal / chemistry*, isolation & purification*, metabolism Cell Line Electrophoresis, Polyacrylamide Gel Polysaccharides / analysis* Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods* |
| Grant Support | |
ID/Acronym/Agency:
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//Biotechnology and Biological Sciences Research Council |
| Chemical | |
Reg. No./Substance:
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0/Antibodies, Monoclonal; 0/Polysaccharides |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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