Chapter title |
Glycosylation Profiling of α/β T Cell Receptor Constant Domains Expressed in Mammalian Cells
|
---|---|
Chapter number | 12 |
Book title |
Synthetic Antibodies
|
Published in |
Methods in molecular biology, March 2017
|
DOI | 10.1007/978-1-4939-6857-2_12 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6855-8, 978-1-4939-6857-2
|
Authors |
Kai Zhang, Stephen J. Demarest, Xiufeng Wu, Jonathan R. Fitchett |
Editors |
Thomas Tiller |
Abstract |
Glycoprofiling recombinant proteins expressed and secreted from mammalian cells is key to understanding their interactions with glycoprotein receptors in vivo. Recently, recombinant T cell receptors (TCRs) are being considered as therapeutic moieties. Here we present a mass spectrometry based protocol with a "bottom up" approach to characterize glycosylation in recombinant fusion proteins with α/β TCR constant domains expressed in mammalian cells. The protocol focuses on using peptide mass mapping and mass spectrometry for N-linked glycan profiling, including analyses of site occupancy, glycan heterogeneity, and possible glycan compositions and structures. |
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