Capillary electrophoresis of sialic acid-containing glycoprotein.: Effect of the heterogeneity of carbohydrate chains on glycoform separation using an α1-acid glycoprotein as a model

被引:48
作者
Kakehi, K [1 ]
Kinoshita, M [1 ]
Kawakami, D [1 ]
Tanaka, J [1 ]
Sei, K [1 ]
Endo, K [1 ]
Oda, Y [1 ]
Iwaki, M [1 ]
Masuko, T [1 ]
机构
[1] Kinki Univ, Fac Pharmaceut Sci, Higashiosaka, Osaka 5778502, Japan
关键词
D O I
10.1021/ac001382u
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
alpha (1)-Acid glycoprotein (AGP) showed multiple peaks on separation using capillary electrophoresis in a chemically modified capillary dth dimethylpolysiloxane at slightly acidic conditions. We analyzed glycoforms of AGP species after separation by ion-exchange chromatography Con A affinity chromatography, and Cu(II)-chelating affinity chromatography. The AGP species thus obtained were digested with N-glycosidase F, and the released carbohydrate chains were analyzed by high-performance liquid chromatography after labeling with 3-aminobenzoic acid. The results afforded basic information on the contribution of carbohydrate chains to the separation mechanism of glycoforms of AGP by capillary electrophoresis, In addition, we describe an easy method for AGP analysis in serum samples using the electrokinetic injection.
引用
收藏
页码:2640 / 2647
页数:8
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