SEPARATION METHODS FOR GLYCOPROTEIN ANALYSIS AND PREPARATION

Several chromatographic methods have been developed for the isolation and characterization of glycoproteins. In these methods, affinity chro matography, a single-step method, or combined use with general chromat ographic methods have now become essential for the purification of man y biologically important glycoproteins, including alpha(1)-acid glycop rotein, immunoglobulins, ceruloplasmin and erythropoietin. On the othe r hand, almost all glycoproteins exhibit polymorphism associated with their glycan moieties. This feature is wide-spread and has been observ ed in natural as well as in recombinant DNA glycoproteins. Recently, s everal sophisticated techniques - such as electromigration method (hig h-performance capillary electrophoresis) and chromatographic methods ( two-dimensional polyacrylamide gel electrophoresis, high-pH anion-exch ange chromatography with pulsed-amperometric detection) - have been in troduced for qualitative or quantitative estimation of the microhetero geneity of glycoproteins. For gaining further insight into the structu re-function relations for microheterogeneity, preparative chromatograp hic techniques that can yield sufficient quantities of glycoprotein va riants must be developed. (C) 1997 Elsevier Science B.V.