LC/MS-based Intact IgG and released glycan analysis for bioprocessing applications.
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Abstract | :
Robust plate based antibody glycan analysis platforms are urgently needed for biopharmaceutical development and manufacturing as well as for clinical biomarker research. A 96-well plate based workflow has been developed to analyze both intact IgG antibodies and released N-glycans using an Orbitrap Fusion Mass Spectrometer and an LC/MS method on the Waters UNIFI platform. Here we describe such a workflow including protein A purification, PNGaseF digestion, 2-AB labeling and SPE clean-up. The measured IgG glycan profile is consistent with that obtained from non-plate based method and commercial kit and has the advantage of less hands-on time. We also demonstrated the application of the workflow in cell culture monitoring and clonal selection work. Apart from checking the major glycan structure changes among clones, post translational modifications such as C- terminal lysine residue clipping and N-terminal pyroglutamic acid formation can also be deduced from the workflow. |
Year of Publication | :
2018
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Journal | :
Biotechnology journal
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Date Published | :
2018
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ISSN Number | :
1860-6768
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URL | :
http://dx.doi.org/10.1002/biot.201700185
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DOI | :
10.1002/biot.201700185
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Short Title | :
Biotechnol J
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