Separation methods for glycoprotein analysis and preparation.
Several chromatographic methods have been developed for the isolation and characterization of glycoproteins. In these methods, affinity chromatography, a single-step method, or combined use with general chromatographic methods have now become essential for the purification of many biologically important glycoproteins, including alpha1-acid glycoprotein, immunoglobulins, ceruloplasmin and erythropoietin. On the other hand, almost all glycoproteins exhibit polymorphism associated with their glycan moieties. This feature is wide-spread and has been observed in natural as well as in recombinant DNA glycoproteins. Recently, several sophisticated techniques--such as electromigration method (high-performance capillary electrophoresis) and chromatographic methods (two-dimensional polyacrylamide gel electrophoresis, high-pH anion-exchange chromatography with pulsed-amperometric detection)--have been introduced for qualitative or quantitative estimation of the microheterogeneity of glycoproteins. For gaining further insight into the structure-function relations for microheterogeneity, preparative chromatographic techniques that can yield sufficient quantities of glycoprotein variants must be developed.
Kishino
S
S
Department of Pharmacy, Hokkaido University Hospital, School of Medicine, Hokkaido University, Sapporo, Japan.
Miyazaki
K
K
D002240:Carbohydrate Sequence; D002570:Ceruloplasmin / Q000737:chemistry / Q000302:isolation & purification; D002853:Chromatography, Liquid / Q000379:methods*; D004586:Electrophoresis / Q000379:methods*; D004921:Erythropoietin / Q000737:chemistry / Q000652:urine; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009961:Orosomucoid / Q000737:chemistry / Q000302:isolation & purification
D006023:Glycoproteins; D007074:Immunoglobulin G; D009961:Orosomucoid; D004921:Erythropoietin; D002570:Ceruloplasmin