The heterogeneous nature of most polysaccharides found in nature includes distribution in molecular weight, primary sequence, and branching. The analytical methodology used in the characterization of these structural aspects must ensure high separation efficiency and selectivity. This paper reports
High Resolution and Rapid Analysis of Branched Oligosaccharides by Capillary Electrophoresis
β Scribed by P. Camilleri; G.B. Harland; G. Okafo
- Publisher
- Elsevier Science
- Year
- 1995
- Tongue
- English
- Weight
- 553 KB
- Volume
- 230
- Category
- Article
- ISSN
- 0003-2697
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β¦ Synopsis
The fluorophore 2-aminoacridone has been used to label a number of branched oligosaccharides previously released from various glycoproteins. Complex glycans were derivatized via their reducing end with this fluorophore by a Schiff's base mechanism followed by reduction to a secondary amine using sodium cyanoborohydride. This process of derivatization was carried out efficiently and in a nonselective manner over a period of 30 min at 90 degrees C. The resulting derivatives were separated with high resolution by capillary electrophoresis using borate buffer, containing taurodeoxycholate, as the separation buffer. The method described does not require the removal of sialic acid residues prior to derivatization, so that treatment of glycans with N-acetyl neuraminidase provided useful and additional structural information. Attempts have also been made to relate the electrophoretic mobility of branched oligosaccharides with their molecular volume.
π SIMILAR VOLUMES
The oligosaccharides in ovalbumin as a glycoprotein model were released with anhydrous hydrazine, and reductively pyridylaminated after re-N-acetylation. The derivatives were analyzed by capillary zone electrophoresis (CZE) with on-column fluorometric detection. Direct CZE could separate the derivat
## Abstract We report on microbore liquid chromatography (ΞΌLC) and capillary electrophoresis (CE) separation of glycopeptides and highβmannoseβtype oligosaccharides, digested from recombinant phospholipaseΒ C, expressed in __Pichia pastoris.__ The glycopeptides were subject to ΞΌLC/electrospray ioniz