title: Frontal affinity chromatography: practice of weak interaction analysis between lectins and fluorescently labeled oligosaccharides.
abstract: Frontal affinity chromatography (FAC) is a simple and effective method that is applicable to the analysis of interactions between glycans and glycan-recognition proteins, including lectins, with weak affinity ranging from 10(-4) to 10(-6) (M) in terms of dissociation constant (Kd). Using conventional instruments, such as a high-performance liquid chromatography (HPLC) system equipped with pump, injector, (fluorescent) detector, and data recorder, the dissociation constants for weak glycan-based interactions can be easily determined with high throughput and accuracy. Notably, if the glycans are labeled with fluorescent dyes, only a small amount of glycans is required for the analysis. Fluorescent labeling of glycans is a common technique, and an increasing number of fluorescent-labeled glycans are commercially available. In this chapter, an advanced FAC method using fluorescent-labeled glycans is described.
authors.affiliation: Bioscience and Biotechnology Center, Nagoya University, Nagoya, 464-8601, Japan, chi@agr.nagoya-u.ac.jp.
authors.forename: Chihiro
authors.initials: C
authors.lastname: Sato
mesh_terms: D002846:Chromatography, Affinity / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D037102:Lectins / Q000378:metabolism*; D009844:Oligosaccharides / Q000737:chemistry* / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding
chemical_list: D005456:Fluorescent Dyes; D037102:Lectins; D009844:Oligosaccharides; D011134:Polysaccharides