Preliminary investigation of glycosylated proteins by capillary electrophoresis and capillary electrophoresis/mass spectrometry using electrospray ionization and by matrix-assisted laser desorption ionization/time-to-flight mass spectrometry.
In recent years, the biotechnological industry has emerged as the major source of new human therapeutic proteins. Although the great majority of these occur naturally as glycoproteins, it has been observed that glycosylation of the recombinantly produced proteins could be fundamental for their in vivo activity (e.g. tissue plasminogen activator, erythropoietin) or, on the contrary, insignificant (e.g. interleukin-1 receptor antagonist, gamma-interferon, granulocyte macrophage-colony-stimulating factor). The inherent heterogeneity of these complex biomolecules presents an exciting challenge in the analytical field for both their structural analysis and the development of suitable analytical methods to guarantee consistency of their production. Owing to this ever increasing therapeutic interest in proteins and glycoproteins, this paper compares the information provided by different analytical techniques (i.e. high-performance liquid chromatography, sodium dodecyl sulphate polyacrylamide gel electrophoresis, capillary electrophoresis, capillary electrophoresis/electrospray ionization mass spectrometry, matrix-assisted laser desorption/ionization time-of flight mass spectrometry and high-performance liquid chromatography/matrix-assisted laser desorption/ionization time-of flight mass spectrometry when used for the analysis of proteins and glycoproteins. For the sake of simplicity, reference standard proteins and glycoproteins were used as samples.
Bonfichi
R
R
Marion Merrell Dow Research Institute, Lepetit Research Centre, Gerenzano (VA), Italy.
Sottani
C
C
Colombo
L
L
Coutant
J E
JE
Riva
E
E
Zanette
D
D
D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000737:chemistry*; D006031:Glycosylation; D007202:Indicators and Reagents; D013058:Mass Spectrometry; D011506:Proteins / Q000737:chemistry*; D012015:Reference Standards; D012996:Solutions; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D006023:Glycoproteins; D007202:Indicators and Reagents; D011506:Proteins; D012996:Solutions