Aptamer-based detection and quantitative analysis of ricin using affinity probe capillary electrophoresis

被引:58
作者
Haes, Amanda J. [1 ]
Giordano, Braden C. [1 ]
Collins, Greg E. [1 ]
机构
[1] USN, Res Lab, Div Chem, Washington, DC 20375 USA
关键词
D O I
10.1021/ac060021x
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
The ability to detect sub-nanomolar concentrations of ricin using fluorescently tagged RNA aptamers is demonstrated. Aptamers rival the specificity of antibodies and have the power to simplify immunoassays using capillary electrophoresis. Under nonequilibrium conditions, a dissociation constant, K-d, of 134 nM has been monitored between the RNA aptamer and ricin A-chain. With use of this free-solution assay, the detection of 500 pM ( similar to 14 ng/mL) or 7.1 amol of ricin is demonstrated. The presence of interfering proteins such as bovine serum albumin and casein do not inhibit this interaction at sub-nanomolar concentrations. When spiked with RNAse A, ricin can still be detected down to 1 nM concentrations despite severe aptamer degradation. This approach offers a promising method for the rapid, selective, and sensitive detection of biowarfare agents.
引用
收藏
页码:3758 / 3764
页数:7
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