| Popis: |
Frontal affinity chromatography (FAC) is a versatile method to analyze biomolecular interactions in terms of dissociation constants (Kds). The application of FAC to lectin-oligosaccharide interaction analysis is particularly important because it is a key process for elucidating the complex features of glycans, the third class of bioinformative macromolecules; global features of glycans remain more elusive than those of nucleic acids and proteins. Considering that an increasing number of lectins as well as carbohydrate-recognition domains/modules have been investigated both via genome search and by chance, systematic analysis of their basic properties, including oligosaccharide specificity, is crucial. Reinforced FAC is a unique method enabling highly reproducible interaction analysis, typically between fluorescently labeled oligosaccharides and immobilized lectins in a high-throughput manner, whereby even weak interactions (e.g., Kd > 10− 4 M) can be determined precisely by a simple procedure with a small amount of labeled oligosaccharides (~ 1 pmol). |
