Sun M, Lee C S
Department of Chemistry and Ames Laboratory, USDOE, Iowa State University, Ames, Iowa 50011, USA.
Biotechnol Bioeng. 1998 Mar 5;57(5):545-51.
The introduction of an enzyme array-electrochemical detection method for carbohydrate analysis is demonstrated by using two complex and one high mannose N-linked oligosaccharides. Instead of measuring the remaining uncleaved oligosaccharides in enzymatic digestion, released monosaccharides are directly quantified by pulsed amperometric detection at a gold electrode. The measured monosaccharide concentrations in combination with the enzyme array analysis provide structural characterization of oligosaccharides. The enzyme array-electrochemical detection method does not require any separation procedure or any prior labeling of oligosaccharides. However, this method is limited by the use of purified oligosaccharide samples and the nature of the enzyme array. The development of more sophisticated enzyme arrays relies upon the introduction of a bank of highly specific (bond, arm, aglycon) exoglycosidases.
通过使用两种复合型和一种高甘露糖型N-连接寡糖,展示了一种用于碳水化合物分析的酶阵列-电化学检测方法。不是测量酶促消化中剩余未切割的寡糖,而是通过在金电极上的脉冲安培检测直接定量释放的单糖。所测量的单糖浓度与酶阵列分析相结合,可提供寡糖的结构表征。酶阵列-电化学检测方法不需要任何分离程序或寡糖的任何预先标记。然而,该方法受到纯化寡糖样品的使用和酶阵列性质的限制。更复杂的酶阵列的开发依赖于引入一组高度特异性(键、臂、糖苷配基)外切糖苷酶。