Frontal affinity chromatography: practice of weak interaction analysis between lectins and fluorescently labeled...

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.