Usefulness of sugar mapping by liquid chromatography/mass spectrometry in comparability assessments of glycoprotein products.
We have previously reported that sugar-mapping by liquid chromatography/mass spectrometry (LC/MS) equipped with a graphitized carbon column (GCC) can be useful for structural analysis of carbohydrates in a glycoprotein. In this paper, we evaluated sugar-mapping with regard to its use in comparability assessment of glycoprotein products. Erythropoietins (EPO) produced from three different sources were chosen as models of the closely related glycoprotein products. The two-dimensional displays of sugar maps drawn by LC/MS with GCC clearly showed the differences in carbohydrate heterogeneity with regard to sialylation, acetylation, and sulphation patterns among three EPOs. Exoglycosidase digestion followed by sugar-mapping provided information regarding the structure of characteristic carbohydrates in each EPO. These results demonstrate that LC/MS with GCC can reveal the details of carbohydrate heterogeneity in order to distinguish between closely related glycoprotein products. Our method can thus be useful in comparability assessments of therapeutic glycoproteins.
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1, Kamiyoga, Setagaya-ku, Tokyo, 158-8501, Japan. nana@nihs.go.jp
Nana
N
Kawasaki
Miyako
M
Ohta
Satsuki
S
Itoh
Masashi
M
Hyuga
Sumiko
S
Hyuga
Takao
T
Hayakawa
D000818:Animals; D016466:CHO Cells; D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D002853:Chromatography, Liquid / Q000379:methods*; D006224:Cricetinae; D004921:Erythropoietin / Q000737:chemistry*; D006023:Glycoproteins / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods*; D009844:Oligosaccharides; D011994:Recombinant Proteins; D013402:Sugar Alcohols / Q000737:chemistry; D013997:Time Factors
D002241:Carbohydrates; D006023:Glycoproteins; D009844:Oligosaccharides; D011994:Recombinant Proteins; D013402:Sugar Alcohols; D004921:Erythropoietin; D006026:Glycoside Hydrolases