Pilot Scale Expression and Purification of Soluble Protein A Tagged β1,6N-Acetylglucosaminyltransferase in CHO Cells
Expression of recombinant soluble protein A tagged mouse core 2 β6-N-acetylglucosaminyltransferase (E.C. 2.4.1.102) has been scaled-up in CHO cells using a continuously operating fluidized bed system yielding 0.3 U/day. A one step 213 fold purification by affinity chromatography on IgG-Sepharose yie...
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| Veröffentlicht in: | Biochemical and biophysical research communications 1997-08, Vol.237 (3), p.653-658 |
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| Hauptverfasser: | , , , , , |
| Format: | Artikel |
| Sprache: | eng |
| Online-Zugang: | Volltext |
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| Zusammenfassung: | Expression of recombinant soluble protein A tagged mouse core 2 β6-N-acetylglucosaminyltransferase (E.C. 2.4.1.102) has been scaled-up in CHO cells using a continuously operating fluidized bed system yielding 0.3 U/day. A one step 213 fold purification by affinity chromatography on IgG-Sepharose yielded a stable enzyme preparation with a specific activity of 44 mU/mg. The enzyme was shown to belong to the L-type with a highly restricted specificity for the acceptor substrate Galβ1→3GalNAcα1→R (core 1). Only little activity towards GlcNAcβ1→3Gal-NAcα1→R (core 3) ( |
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| ISSN: | 0006-291X 1090-2104 |
| DOI: | 10.1006/bbrc.1997.7209 |