On the preparative use of recombinant pig α(1-3)galactosyl-transferase

Bioorganic and Medicinal Chemistry Letters

Volumn 8, Issue 12, 1998, Pages 1575-1578

  Baisch, Gabi ^a   Öhrlein, Reinhold ^a   Kolbinger, Frank ^a   Streiff, Markus ^a  

^a NOVARTIS PHARMA AG   (Switzerland)

Author keywords

[No Author keywords available]

Indexed keywords

GALACTOSE; GALACTOSYLTRANSFERASE; LACTOSE; RECOMBINANT ENZYME; TRISACCHARIDE; URIDINE DIPHOSPHATE GALACTOSE;

ANIMAL TISSUE; ARTICLE; CONTROLLED STUDY; DRUG SYNTHESIS; GLYCOSYLATION; MASS SPECTROMETRY; NONHUMAN; NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY; SWINE;

ANIMALS; GALACTOSYLTRANSFERASES; GLYCOSYLATION; MAGNETIC RESONANCE SPECTROSCOPY; RECOMBINANT PROTEINS; SWINE;

ANIMALIA; SUS SCROFA;

EID: 0032537524     PISSN: 0960894X     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0960-894X(98)00266-2     Document Type: Article

References (17)

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12. 13. For large scale expression the fusion protein was recloned into Baculovirus. The secreted ProteinA-α(1-3)gal-t fusion protein was harvested 3 days post-infection from the medium of insect cells and purified using SP- and Q-Sepharose followed by affinity chromatography on UDP-hexanolamine-Sepharose.
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16. 2. This mixture is briefly vortexed and then incubated at 37°C with 125 μ1 of a galactosyl-transferase solution (3U/ml) and 3 μ1 calf intestine alkaline phosphatase (Boehringer no. 108146, 7500 U/498 μ1) for 48 h. The turbid mixture is then passed over a short C-18 reversed phase column, washed with water and eluted with methanol. The organic phase is evaporated and the residue purified on silica gel (eluent: dichloromethane - methanol - water / 10 - 2 - 0.2) to give 7.2 mg (64 %) of the title compound as a white powder after a final lyophilization from dioxane-water.
17. 17) Recently the parent compound (R = Ac) has been prepared in a one-pot reaction with both β(1-4)- and α(1-3)galactosyl-transferase on a microscale: Hokke, C. H.; Zervosen, A.; Elling, L.; Joziasse, D. H.; van den Eijnden, D. H. Glycoconjugate J., 1996, 13, 687.