Abstract: Methods for profiling oligosaccharides released from glycoproteins and methods for the electrophoretic separation of high mannose, complex and hybrid oligosaccharides are disclosed. The methods include first providing an analytical sample which includes at least one released oligosaccharide and introducing the sample into an electrophoretic channel containing a separation medium having a pH less than 8 and greater than 2.5. Then applying a sufficiently high electric field causes the released oligosaccharides to migrate and separate within the electrophoretic channel in accordance with their molecular size or charge to mass ratio. Detecting the separated oligosaccharides provides glycoprotein conjugate profiling information. Preferred embodiments include using a separation medium which includes lithium acetate buffer having a pH of about 4.75 and a gel of about 0.4% polyethylene oxide.

Abstract: Methods for profiling oligosaccharides released from glycoproteins and methods for the electrophoretic separation of high mannose, complex and hybrid oligosaccharides are disclosed. The disclosed methods include first providing an analytical sample which includes at least one released oligosaccharide and introducing the sample into an electrophoretic channel containing an electrophoretic separation medium having a pH less than 8 and greater than 2.5.Then applying a sufficiently high electric field causes the released oligosaccharides to electrophoretically migrate and separate within the electrophoresis channel in accordance with their molecular size or charge to mass ratio. Detecting the separated oligosaccharides provides glycoprotein conjugate profiling information. Preferred embodiments include using an electrophoresis separation medium which includes a lithium acetate buffer having pH of about 4.75 and a gel of about 0.4 wt % polyethylene oxide.