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 (K d). 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.
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Sato, C. (2014). Frontal Affinity Chromatography: Practice of Weak Interaction Analysis Between Lectins and Fluorescently Labeled Oligosaccharides. In: Hirabayashi, J. (eds) Lectins. Methods in Molecular Biology, vol 1200. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1292-6_22
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DOI: https://doi.org/10.1007/978-1-4939-1292-6_22
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