PROBING THE CARBOHYDRATE SIDE CHAINS OF RECOMBINANT TISSUE PLASMINOGEN ACTIVATOR
Abstract
The glycosylation of recombinant tissue plasminogen activator derived from transfected CHO cells was assessed using glycosyltransferases to probe for terminal Gal and GlcNAc residues and solid-state lectin binding assays. Analysis of the oligosaccharides released from each glycosylation site by hydrazinolysis by Con A affinity chromatography/ anion exchange HPLC/ BioGel P4 gel filtration corroborated the glycosyltransferase and lectin-binding assay data. Significant undersialylation of the oligosaccharides of t-PA and less than 1% of free terminal GlcNAc residues was indicated. The above assays could be useful in monitoring glycosylation status during quality control in recombinant glycoprotein production.Citation
Protein glycosylation, 275 - 278Affiliation
MRC Collaborative Centre, 1-3 Burtonhole Lane, Mill Hill, London NW7 1AD and National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, U.K.Type
Book chapterconference paper
Language
enSeries/Report no.
GBF monographs ; Volume 15ISSN
0930-4320ISBN
15608118463527283676
Collections
The following license files are associated with this item:
- Creative Commons
Except where otherwise noted, this item's license is described as Attribution-NonCommercial-ShareAlike 4.0 International