Aptamers offer many advantages that make them very attractive to the development of detection and diagnostic tools from health to food science. Here, we are introducing a novel aptamer-based biosensor that can be used for detecting RNase contamination or foodborne pathogens with further configuration. This biosensor is composed of a RNA aptamer combined with a conditional fluorescent molecule that loses fluorescence when detached from the aptamer, after RNA aptamer degradation. The characterization of aptamer-based biosensor is a labor-intensive and time-consuming process when using gel electrophoresis-based methods. It also requires the accurate repetition of the generation of wide range concentration gradient. However, generation of a wide range of concentration gradient requires large amount of rare and precious samples and can hardly be obtained with conventional methodology. Therefore, we used our integrated microfluidic platform for gradient formation that we previously described (Anal Chem 2011, 83 (16):6148-6153).To characterize the degradation behavior of aptamer-based biosensor, we generated a log-scale concentration gradient of degrading agents: lead acetate, RNase T1, or RNase A, using our microfluidic gradient generator system. We measured the fluorescence intensity to quantify the degradation of biosensor over different times and we obtained the degradation profiles of biosensor. The different degradation behaviors of biosensor by degrading agents were observed and the degradation profiles can be used to differentiate degrading agents due to the different degradation kinetics. Our microfluidic method can provide rapid characterization of our aptamer-based biosensor within minutes while gel electrophoresis-based methods require at least several hours.

Rapid characterization of a novel aptamer-based biosensor on a chip

247th National Meeting (2014)

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