title: Partial-filling affinity capillary electrophoresis of glycoprotein oligosaccharides derivatized with 8-aminopyrene-1,3,6-trisulfonic acid.
abstract: Partial-filling affinity capillary electrophoresis has been applied to the simultaneous analysis of interactions between glycoprotein oligosaccharides and certain plant lectins. A lectin solution and a mixture of glycoprotein-derived oligosaccharides labeled with 8-aminopyrene-1,3,6-trisulfonic acid were introduced to a neutrally coated capillary in this order, and separated by application of a negative voltage. Interaction of a lectin with each oligosaccharide in the mixture was observed as the specific retardation or dissipation of peaks, in addition to the size/charge separation of oligosaccharides by zone electrophoresis in the remainder (≈90%) of the capillary. The strength of the interaction with lectin was controlled by introducing an appropriate volume of lectin solution. Application of various specificities of lectins indicated characteristic migration profiles of the oligosaccharides. Moreover, sequential injection of four lectins (Maachia amurensis mitogen, Sambucus sieboldiana agglutinin, Erythrina cristagalli agglutinin, Aleuria aurantia lectin) induced complete dissipation of complex-type oligosaccharides and enabled specific determination of the presence of high-mannose oligosaccharides without the interference or alteration of the electropherogram in porcine thyroglobulin. This method was also applied to determine the binding constants of ovalbumin-derived oligosaccharides to wheat germ agglutinin.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kinki University, 3-4-1 Kowakae, Higashi-Osaka, Japan.
authors.forename: Sachio
authors.initials: S
authors.lastname: Yamamoto
authors.forename: Chikayo
authors.initials: C
authors.lastname: Shinohara
authors.forename: Eriko
authors.initials: E
authors.lastname: Fukushima
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Kakehi
authors.forename: Takao
authors.initials: T
authors.lastname: Hayakawa
authors.forename: Shigeo
authors.initials: S
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D019075:Electrophoresis, Capillary / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D009844:Oligosaccharides / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D037121:Plant Lectins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding; D011721:Pyrenes / Q000737:chemistry*; D013552:Swine
chemical_list: C103430:8-aminopyrene-1,3,6-trisulfonic acid; D006023:Glycoproteins; D009844:Oligosaccharides; D037121:Plant Lectins; D011721:Pyrenes