Selective isolation of Vibrio cholerae neuraminidase using an immobilized 4-(nitrophenyl)oxamic acid.
N-(4-Nitrophenyl)oxamic acid[1] (1) was coupled with Sepharose 4B containing 1,6-diaminohexane as spacer group. This material was used as a specific adsorbent in the purification of Vibrio cholerae neuraminidase. The enzyme was completely retarded and separated from the bulk of the protein when washed with 50mM sodium acetate buffer, pH 5.0. A stepwise increase of sodium chloride concentration from 1.0 to 2.0M was found to be necessary for a sharp elution of neuraminidase activity. The purification was tenfold, and a recovery of more than 90% was obtained. Neuraminidase is only weakly retarded on a column of 1,6-diaminohexane coupled with Sepharose 4B and is not adsorbed by Sepharose 4B.
D002846:Chromatography, Affinity; D007700:Kinetics; D009439:Neuraminidase / Q000302:isolation & purification* / Q000378:metabolism; D009596:Nitrophenols; D009994:Osmolar Concentration; D010072:Oxamic Acid / Q000031:analogs & derivatives; D011485:Protein Binding; D012965:Sodium Chloride / Q000494:pharmacology; D014734:Vibrio cholerae / Q000201:enzymology*
D009596:Nitrophenols; D012965:Sodium Chloride; D009439:Neuraminidase; D010072:Oxamic Acid