A pico-HPLC-LIF system for the amplification-free determination of multiple miRNAs in cells |
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| Authors: | Wenmei Zhang Zunsheng Han Yingqi Liang Qi Zhang Xiangnan Dou Guangsheng Guo Xiayan Wang |
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| Institution: | 1. Center of Excellence for Environmental Safety and Biological Effects, Beijing Key Laboratory for Green Catalysis and Separation, Department of Chemistry and Biology, Beijing University of Technology, Beijing 100124, China;2. State Key Laboratory of Bioactive Substance and Function of Natural Medicines, Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China |
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| Abstract: | MicroRNAs are a class of important biomarkers,and the simultaneous detection of multiple miRNAs can provide valuable information about many diseases and biological processes.Amplification-free determination has been developed for the analysis of multiple miRNAs because of its characteristic low cost and high fidelity.Herein,a method for the amplification-free analysis and simultaneous detection of multiple miRNAs based on a so-called pico-HPLC-LIF system is described.In this process,a bare open capilla ry with an inner diameter of 680 nm is used as a sepa ration column for a sample volume of several hundreds of femtoliters(300 fL),followed by separation and detection.The technique has a zeptomolar limit of detection.The method was applied to detect cellular miRNA from adenocarcinomic human alveolar basal epithelial(A549) cell extracts,and the simultaneous detection of the mir-182,miR-155,and let-7 a was achieved.The results showed that the expression of mir-182 and miR-155 was up-regulated and that of let-7 a was down-regulated in A549 cells.This method for multiple miRNAs detection is expected to have broad applications in miRNA-based disease diagnosis,prognosis,treatment,and monitoring. |
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| Keywords: | Multiple miRNAs Amplification-free Pico-HPLC Nanocapillary Laser-induced fluorescence |
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