Lectin affinity electrophoresis.
Lectin affinity electrophoresis is a powerful technique to investigate the interaction between a lectin and its ligand. Affinity electrophoresis results from the reduced mobility of a charged species owing to its interaction with an immobile species. In this protocol, a two-dimensional lectin affinity electrophoresis experiment is described that affords separation of oligosaccharides. The first-dimension is composed of a weak, polyacrylamide, capillary tube gel containing a lectin. The example described involves a mixture of fluorescently labeled disaccharides. The mobility of only the lectin-binding disaccharide is reduced affording a separation in the first-dimension. The tube gel is then extruded and placed onto the second-dimension gradient polyacrylamide gel and subjected to electrophoresis. Mobility in the second-dimension is dependent on molecular size and visualization si by fluorescence under transillumination. This method is also applicable, with appropriate modifications, for the separation and analysis of glycopeptides and glycoproteins.
Linhardt
R J
RJ
Division of Medical and Natural Products Chemistry, College of Pharmacy, University of Iowa, Iowa City 52242, USA.
Han
X J
XJ
Fromm
J R
JR
D002240:Carbohydrate Sequence; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006020:Glycopeptides / Q000032:analysis; D037102:Lectins / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009844:Oligosaccharides / Q000032:analysis; D037121:Plant Lectins; D010944:Plants / Q000737:chemistry
D006020:Glycopeptides; D037102:Lectins; D009844:Oligosaccharides; D037121:Plant Lectins