N-Acetyl-Neuraminic Acid → 3-O-(N-Acetyl-alpha-neuraminosyl)-beta-D-galactopyranose

Participants

Role	Name	HCID	Charge
Reactant	N-Acetyl-Neuraminic Acid	HCID 439197	—
Reactant	2-(acetylamino)-2-deoxy-3-O-beta-D-galactopyranosyl-D-galactopyranose	HCID 441248	—
Reactant	CMP-Neu5Ac	HCID 448209	—
Reactant	(2R,3S,4S,5R)-2,3,4,5,6-pentahydroxyhexanal	HCID 3037556	—
Reactant	beta-D-Galp-(1->3)-beta-D-GlcpNAc	HCID 10177989	—
Reactant	sodium;[5-acetamido-2-carboxy-4-hydroxy-6-(1,2,3-trihydroxypropyl)oxan-2-yl] [(2R,3S,4R,5R)-5-(4-amino-2-oxopyrimidin-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate	HCID 16760374	—
Product	3-O-(N-Acetyl-alpha-neuraminosyl)-beta-D-galactopyranose	HCID 5289366	—

Reaction Steps

The resulting CMP-sialic acid analogue (which in FIG. 3 is illustrated as the CMP derivative of Neu5Ac, i.e., CMP-Neu5Ac) is then combined with the derivatized βGal(1→3)βGlcNAc-OR compound in the presence of the βGal(1→3/4)βGlcNAc α(2→3)sialyltransferase or the βGal(1→3)GalNAc α(2→3)sialyltransferase under conditions wherein sialic acid is transferred to the 3 position of the galactose to form a αNeu5Ac(2→3)βGal linkage. Suitable conditions, known in the art, include the addition of the sialyltransferase to a mixture of the derivatized βGal(1→3)βGlcNAc-OR compound and of the CMP-sialic acid in a appropriate buffer such as 0.1M sodium cacodylate in appropriate conditions of pH and temperature such as at a pH of 6.5 to 7.5 and a temperature between 25° and 45° C., preferably 35°-40° C., while incubating for 12 hours to 4 days. The resulting sialylated product can be isolated and purified using conventional methodology comprising HPLC, ion exchange-, gel-, reverse-phase- or adsorption chromatography.

Provenance & Evidence

DOI	10.6084/m9.figshare.5104873.v1
Patent number	US05872096
Source dataset	uspto-grants-1999_02
ORD ID	ord-857116f0617b473b8f8deecd872b47d2
Source note	uspto-grants-1999_02