SpringerProtocols: Abstract: 13. Use of Capillary Electrophoresis for Polysaccharide Studies and Applications

13. Use of Capillary Electrophoresis for Polysaccharide Studies and Applications

By: Amelia Gamini³, Anna Coslovi³, Isabella Rustighi³, Cristiana Campa⁴, Amedeo Vetere³, Sergio Paoletti³

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Capillary electrophoresis (CE) applications to charged polysaccharides are briefly reported. A simple procedure is presented to determine the esterification degree of a hyaluronan derivative. In this case, the degree of substitution was as low as 14%. The molecular weight distribution of mannuronic oligosaccharides mixture produced by hydrolysis of native polymannuronic is readily calculated from peak area of the species resolved by CE on the basis of a specific degree of polymerization. The influence of the applied electric field strength on the free solution mobility of hyaluronan samples is briefly addressed for molar masses of the order of 10⁵ and 10⁶ g/mol. The data are compared with the results obtained for a 50% galactose-substituted hyaluronic acid (HA). Mobility data obtained as a function of buffer pH for a native HA sample as well as for two galactose-amide HA derivatives, having slightly different degrees of substitution, are presented and discussed in terms of the polymer charge density parameters ξ. In most cases, more questions than answers arise from the application of CE to charged polysaccharides. However, perspectives are disclosed for a further understanding of the reliability of CE applied for the structural elucidation of such macromolecules.

Affiliation(s): (3) Department of Biochemistry, Biophysics and Macromolecular Chemistry, University of Trieste, Trieste, Italy
(4) Novartis Vaccines and Diagnostics, Siena, Italy

Book Title: Capillary Electrophoresis: Methods and Protocols

Series: Methods in Molecular Biology | Volume: 384 | Pub. Date: Jan-04-2008 | Page Range: 357-400 | DOI: 10.1007/978-1-59745-376-9_13

Subject: Cell Biology

Key Words: Charged polysaccharides - hyaluronan - capillary electrophoresis - electrophoretic mobility - charge density - molar mass distribution - random degradation - glycoconjugates

References

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