Quantitation of surface-reducing-end covalently bound polysaccharides via hydrazinolysis and deamination.
Depending on the method of deposition, reactive sites of polysaccharides on substrates may not be available when their reducing ends have been used to covalently bind them to the substrates. Here we present a method that allows surface density measurements of reducing-end covalently bound polysaccharides in a procedure that cleaves the polysaccharide chain from the surface via hydrazinolysis and deamination, leaving on the surface a disaccharide that is later radiolabeled with an aldehyde in a reaction with enamine formation. The method described has the advantage that it may be used with any polysaccharide patterned to any surface exposing an amino-terminated monolayer by reductive amination of their galactosamine or glucosamine repeating units. We illustrate the technique with the quantitation of glycosaminoglycans (GAGs) on silanized glass surfaces.
Peramo
Antonio
A
Department of Physics, UniVersity of South Florida, Tampa, Florida 33620, USA. aperamo@umich.edu
Matthews
Garrett
G
D000447:Aldehydes / Q000737:chemistry; D001671:Biochemistry / Q000379:methods*; D005898:Glass; D006025:Glycosaminoglycans / Q000737:chemistry*; D006834:Hydrazines / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008956:Models, Chemical; D011134:Polysaccharides / Q000737:chemistry*; D012821:Silanes / Q000737:chemistry; D013499:Surface Properties
D000447:Aldehydes; D006025:Glycosaminoglycans; D006834:Hydrazines; D011134:Polysaccharides; D012821:Silanes