Large-scale preparation, purification, and characterization of hyaluronan oligosaccharides from 4-mers to 52-mers.
Hyaluronan (HA) was depolymerized by partial digestion with testicular hyaluronidase and separated into size-uniform HA oligosaccharides from 4-mers to 52-mers by anion exchange chromatography after removal of the hyaluronidase. The purity and size of each HA oligosaccharide was confirmed by using HPLC analyses, FACE, and ESI-MS. (1)H and (13)C NMR assignments and elemental analyses were obtained for each HA oligosaccharide. Endotoxins, proteins, and DNA were absent or in trace amounts in these HA oligosaccharides. Gram/mg-scale hyaluronan oligosaccharides were obtained from 200 g of HA starting material. These pure, size-uniform, and large range of HA oligosaccharides will be available for investigating important biological functions of HA, such as for the determination of the size(s) of HA oligosaccharides that induce angiogenesis or mediate inflammatory responses, and to interact with HA-binding proteins and receptors both in in vitro and in vivo studies.
Tawada
Akira
A
Seikagaku Corporation, Central Research Laboratories, 1253, Tateno 3-chome, Higshiyamato-shi, Tokyo 207-0021, Japan.
Masa
Takahiro
T
Oonuki
Yoji
Y
Watanabe
Atsushi
A
Matsuzaki
Yuji
Y
Asari
Akira
A
D002851:Chromatography, High Pressure Liquid; D004586:Electrophoresis / Q000379:methods; D006820:Hyaluronic Acid / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification; D009844:Oligosaccharides / Q000138:chemical synthesis / Q000737:chemistry* / Q000302:isolation & purification; D021241:Spectrometry, Mass, Electrospray Ionization
D009844:Oligosaccharides; D006820:Hyaluronic Acid