Affinity chromatography of neoglycoproteins.
Glycoproteins, as a class of biomolecules, exhibit much more heterogeneous structures than non-glycosylated proteins. They present a challenging area of research. Model glycoproteins with well-defined protein and carbohydrate structures are helpful in the search for high-resolution methods for the separation of glycoproteins. Neoglycoproteins, maltose-modified chymotrypsin and lactose-modified chymotrypsin, were synthesised by modifying chymotrypsin with maltose and lactose, respectively, using the reductive amination method. Boronate chromatography was applied to isolate the neoglycoproteins from non-glycosylated substances. The use of Tris-HCl as a shielding reagent during the boronate chromatography proved to be efficient in eliminating unwanted interactions between the boronate ligand and the peptide backbone of chymotrypsin. The retention time of neoglycoproteins on the boronate column was increased with increasing the degree of modification.
Li
Y
Y
Gramineer International AB, IDEON, Lund, Sweden.
Larsson
E L
EL
Jungvid
H
H
Galaev IYu
Mattiasson
B
B
D001896:Boron Compounds; D002846:Chromatography, Affinity / Q000379:methods*; D002918:Chymotrypsin / Q000737:chemistry*; D006023:Glycoproteins / Q000302:isolation & purification*; D007785:Lactose; D008320:Maltose
D001896:Boron Compounds; D006023:Glycoproteins; D008320:Maltose; D002918:Chymotrypsin; D007785:Lactose