CAPILLARY GEL-ELECTROPHORESIS SEPARATION OF HIGH-MANNOSE TYPE OLIGOSACCHARIDES DERIVATIZED BY 1-AMINOPYRENE-3,6,8-TRISULFONIC ACID

In this paper we report the capillary gel electrophoresis separation o f 1-aminopyrene-3,6,8-trisulfonic acid (APTS) labeled oligosaccharides , released enzymatically from bovine pancreatic ribonuclease B. The re leased and labeled high-mannose structures were identified by spiking the separated peaks with the appropriate commercially available indivi dual oligosaccharides. Baseline separation of the three positional iso mers of the mannose-7 and mannose-8 oligosaccharides was attained. Com parison of the electrophoretic mobilities of the high-mannose type bra nched carbohydrates to the linear molecules of maltooligosaccharides ( glucose oligomers) have been shown using different gel concentrations in the running buffer system. We observed that increasing gel concentr ation in the running buffer causes an increase in the relative mobilit y values of the high-mannose type carbohydrate molecules compared to t he linear glucose oligomers. Analysis of our data indicated that this increase in relative migration time was not due to sieving, but seemed to be related to the mannose content and hydrodynamic volume of the b ranched glycans as well as to the viscosity of the separation medium.