Synthesis and evaluation of N-acetylneuraminic acid-based affinity matrices for the purification of sialic acid-recognizing proteins.
The synthesis of 2-S-(2-aminoethyl) 5-acetamido-3,5-dideoxy-2-thio-D-galacto-2-nonulopyranosidonic acid (1) has been successfully achieved from the precursors methyl 5-acetamido-4,7,8,9-tetra-O-acetyl-2-S-acetyl-3,5-dideoxy-2-thio-D-glyce ro-alpha-D-galacto-2-nonulopyranosonate (2) and 2-bromo-N-(tert-butoxycarbonyl)-ethylamine (5). Compounds 1 and 2 were coupled, via amino and thioglycosidic linkages, respectively, to epoxy-activated Sepharose 6B. The resultant affinity adsorbents have proved efficient in purifying the sialic acid-recognizing enzyme Vibrio cholerae sialidase, in a one-step process with yields in the order of 60%.
Ciccotosto
S
S
Department of Medicinal Chemistry, Monash University (Parkville Campus), Victoria, Australia.
Kiefel
M J
MJ
Abo
S
S
Stewart
W
W
Quelch
K
K
von Itzstein
M
M
D000345:Affinity Labels / Q000737:chemistry*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D005069:Evaluation Studies as Topic; D009682:Magnetic Resonance Spectroscopy; D013058:Mass Spectrometry / Q000379:methods; D015394:Molecular Structure; D019158:N-Acetylneuraminic Acid / Q000737:chemistry* / Q000378:metabolism*; D009439:Neuraminidase / Q000737:chemistry / Q000302:isolation & purification*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification; D012685:Sepharose / Q000737:chemistry; D014734:Vibrio cholerae / Q000201:enzymology
D000345:Affinity Labels; D011994:Recombinant Proteins; D012685:Sepharose; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid