Exploring enzyme amplification to characterize specificities of protein-carbohydrate recognition.

Publisher Summary A considerable amount of evidence has accumulated indicating that a large number of glycoconjugates become altered during normal and abnormal cellular development. Understanding these diverse biological processes at the molecular level requires knowledge of the structure and biochemistry of cellular oligosaccharides. Because of their exquisite carbohydrate specificity, ready availability, and diversity lectins mostly from plants have become indispensable tools in biological research for the investigation of sugars of glycoconjugates. This chapter describes the principle and procedure employed for enzyme-linked lectinsorbent assay (ELLA) for measuring the carbohydrate-binding activity of lectins using a microtiter assay. The procedure is broadly divided into direct and indirect methods and is discussed with representative lectin examples optimized in author's laboratory. ELLA provides a sensitive biochemical assay with high specificity and broad flexibility to determine the sugar-binding affinities for different classes of lectins and has been applied to various protein–carbohydrate interaction studies

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