Duong Hong Dinh, Rhee Jong Il
School of Applied Chemical Engineering, Research Center for Biophotonics, Chonnam National University, Yong-Bong dong 300, 500-757 Gwangju, Republic of Korea.
Anal Chim Acta. 2008 Sep 19;626(1):53-61. doi: 10.1016/j.aca.2008.07.042. Epub 2008 Aug 3.
In this work, urea detection techniques based on the pH sensitivity of CdSe/ZnS QDs were developed using three types of sol-gel membranes: a QD-entrapped membrane, urease-immobilized membrane and double layer consisting of a QD-entrapped membrane and urease-immobilized membrane. The surface morphology of the sol-gel membranes deposited on the wells in a 24-well microtiter plate was investigated. The linear detection range of urea was in the range of 0-10mM with the three types of sol-gel membranes. The urea detection technique based on the double layer consisting of the QD-entrapped membrane and urease-immobilized membrane resulted in the highest sensitivity to urea due to the Michaelis-Menten kinetic parameters. That is, the Michaelis-Menten constant (K(m)=2.0745mM) of the free urease in the QD-entrapped membrane was about 4-fold higher than that (K(m)=0.549mM) of the immobilized urease in the urease-immobilized membrane and about 12-fold higher than that (K(m)=0.1698mM) of the immobilized urease in the double layer. The good stability of the three sol-gel membranes for urea sensing over 2 months showed that the use of sol-gel membranes immobilized with QDs or an enzyme is suitable for biomedical and environmental applications.
在这项工作中,基于CdSe/ZnS量子点的pH敏感性开发了尿素检测技术,使用了三种类型的溶胶-凝胶膜:量子点包埋膜、固定化脲酶膜以及由量子点包埋膜和固定化脲酶膜组成的双层膜。研究了沉积在24孔微量滴定板孔中的溶胶-凝胶膜的表面形态。这三种类型的溶胶-凝胶膜对尿素的线性检测范围为0-10mM。基于由量子点包埋膜和固定化脲酶膜组成的双层膜的尿素检测技术,由于米氏动力学参数,对尿素具有最高的灵敏度。也就是说,量子点包埋膜中游离脲酶的米氏常数(K(m)=2.0745mM)比固定化脲酶膜中固定化脲酶的米氏常数(K(m)=0.549mM)高约4倍,比双层膜中固定化脲酶的米氏常数(K(m)=0.1698mM)高约12倍。这三种溶胶-凝胶膜在2个月以上的尿素传感过程中具有良好的稳定性,表明使用固定有量子点或酶的溶胶-凝胶膜适用于生物医学和环境应用。