Use of monomeric, monovalent lectin derivatives for flow cytometric analysis of cell surface glycoconjugates

Because of their unique carbohydrate binding characteristics, lectins serve as invaluable tools in biological and medical research; separation and characterization of glycoproteins and glycopeptides, histochemistry of cells and tissues, and the study of cell differentiation (1). Direct analysis of individual cells carrying glycoconjugate on the cell surface by flow cytometry/cell sorting should give invaluable information on the distribution, dynamics, and biological role of these glycoconjugates. However, native lectins with multiple binding sites often suffer from their property to agglutinate cells in these applications. Single cell suspensions are required for these applications to avoid the stacking problems and also for accurate measurement. To attain this, lectins should be used in the concentration range, in which the agglutination does not occur (2). This has been one of the major problems for the application of lectins in this field.