Centre for Research in Biosciences, Faculty of Health and Life Sciences, University of the West of England, Frenchay Campus, Bristol, UK.
Anal Biochem. 2013 Apr 15;435(2):114-9. doi: 10.1016/j.ab.2013.01.006. Epub 2013 Jan 16.
The development of a disposable amperometric biosensor for the measurement of circulating galactose in serum is described. The biosensor comprises a screen-printed carbon electrode (SPCE), incorporating the electrocatalyst cobalt phthalocyanine (CoPC), which is covered by a permselective cellulose acetate (CA) membrane and a layer of immobilized galactose oxidase (GALOX). The optimal response of the biosensor, designated as GALOX-CA-CoPC-SPCE, was obtained by systematically examining the effects of enzyme loading, temperature, pH, and buffer strength. The optimal performance of the biosensor occurred with 2U of GALOX, at 35°C, using 50mM phosphate buffer solution (pH 7.0). The sensitivity was 7.00μAmM(-1)cm(-2) and the linear range from 0.1 to 25mM with a calculated limit of detection (LOD) of 0.02mM; this concentration range and LOD are appropriate to diagnose galactosemia, i.e., concentrations >1.1mM in infants. When the biosensor was used in conjunction with amperometry in stirred solution for the analysis of serum, the precision values obtained on unspiked (endogenous level of 0.153mM) and spiked serum (1mM added) (n=6) were 1.10% and 0.11%, respectively, with a calculated recovery of 99.9%.
描述了一种用于测量血清中循环半乳糖的一次性安培生物传感器的开发。该生物传感器由一个丝网印刷碳电极（SPCE）组成，其中包含电催化剂酞菁钴（CoPC），其被一层具有选择透过性的醋酸纤维素（CA）膜和一层固定化半乳糖氧化酶（GALOX）覆盖。通过系统地研究酶负载、温度、pH 值和缓冲液强度的影响，确定了生物传感器的最佳响应，命名为 GALOX-CA-CoPC-SPCE。在 2U 的 GALOX、35°C 下、使用 50mM 磷酸盐缓冲溶液（pH7.0）时，生物传感器的性能最佳。其灵敏度为 7.00μAmM(-1)cm(-2)，线性范围为 0.1 至 25mM，计算得出的检测限（LOD）为 0.02mM；这个浓度范围和 LOD 适用于诊断半乳糖血症，即婴儿体内浓度>1.1mM。当将该生物传感器与搅拌溶液中的安培法一起用于血清分析时，对未加标（内源性浓度为 0.153mM）和加标血清（添加 1mM）（n=6）的精密度值分别为 1.10%和 0.11%，计算得出的回收率为 99.9%。
