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Application of Fluorophore-Assisted Carbohydrate Electrophoresis to Analysis of Disaccharides and Oligosaccharides Derived From Glycosaminoglycans

Application of Fluorophore-Assisted Carbohydrate Electrophoresis to Analysis of Disaccharides and Oligosaccharides Derived From Glycosaminoglycans

Yoji Oonuki, Yuko Yoshida, Yasuo Uchiyama, Akira Asari

Anal Biochem. 2005 Aug 15;343(2):212-22.

PMID: 16004957

Abstract:

Various combinations of fluorescent dyes, polyacrylamide gels, and electrophoresis buffers were tested by fluorophore-assisted carbohydrate electrophoresis (FACE) for the purpose of analyzing sulfated and nonsulfated glycosaminoglycan (GAG) oligosaccharides in which disaccharides and low-molecular weight oligosaccharides were included. A nonionic fluorescent dye was found to be suitable for analyzing sulfated disaccharides derived from sulfated GAGs (e.g., chondroitin sulfate, dermatan sulfate) because sulfated disaccharides themselves had enough anionic potential for electrophoresis. The migration rates of chondroitin sulfate (CS) disaccharides in polyacrylamide gels were affected by the number of sulfate residues and the conformation of each disaccharide. When an anionic fluorescent dye, 8-aminonaphthalene-1,3,6-trisulfonic acid disodium salt (ANTS), was coupled with sulfated GAG oligosaccharides, nearly all of the conjugates migrated at the electrophoretic front due to the added anionic potential. Nonsulfated hyaluronan (HA) oligosaccharides (2-16 saccharides) were subjected to electrophoresis by coupling with a nonionic fluorescent dye, 2-aminoacridone (AMAC), but did not migrate in the order of their molecular size. Especially di-, tetra-, hexa-, and octasaccharides of HA migrated in the reverse order of their molecular size. HA/CS oligosaccharides were able to migrate in the order of their chain lengths by coupling with an anionic fluorescent dye in a nonborate condition.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
AP5398345	8-Aminonaphthalene-1,3,6-trisulfonic acid disodium salt		5398-34-5	Price

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