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Label-free detection of DNA-binding proteins based on microfluidic solid-state molecular beacon sensor

Wang, Jun ; Onoshima, Daisuke ; Aki, Michihiko ; Okamoto, Yukihiro ; Kaji, Noritada ; Tokeshi, Manabu ; Baba, Yoshinobu

Analytical chemistry, 01 May 2011, Vol.83(9), pp.3528-32 [Peer Reviewed Journal]

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  • Title:
    Label-free detection of DNA-binding proteins based on microfluidic solid-state molecular beacon sensor
  • Author: Wang, Jun ; Onoshima, Daisuke ; Aki, Michihiko ; Okamoto, Yukihiro ; Kaji, Noritada ; Tokeshi, Manabu ; Baba, Yoshinobu
  • Subjects: Biosensing Techniques -- Instrumentation ; DNA Probes -- Chemistry ; DNA-Binding Proteins -- Metabolism ; Microfluidic Analytical Techniques -- Methods ; Oligonucleotide Probes -- Chemistry
  • Is Part Of: Analytical chemistry, 01 May 2011, Vol.83(9), pp.3528-32
  • Description: A solid-state molecular beacon using a gold support as a fluorescence quencher is combined with a polydimethylsiloxane (PDMS) microfluidic channel to construct an optical sensor for detecting single-stranded DNA binding protein (SSBP) and histone protein. The single-stranded DNA-Cy3 probe or double-stranded DNA-Cy3 probe immobilized on the gold surface is prepared for the detection of SSBP or histone, respectively. Due to the different quenching ability of gold to the immobilized single-stranded DNA-Cy3 probe and the immobilized double-stranded DNA-Cy3 probe, low fluorescence intensity of the attached single-stranded DNA-Cy3 is obtained in SSBP detection, whereas high fluorescence intensity of the attached double-stranded DNA-Cy3 is obtained in histone detection. The amounts of SSBP in sample solutions are determined from the degree of fluorescence recovery of the immobilized single-stranded DNA-Cy3 probe, whereas that of histone in sample solutions is determined from the degree of fluorescence quenching of the immobilized double-stranded DNA-Cy3 probe. Using this approach, label-free detection of target proteins at nanomolar concentrations is achieved in a convenient, general, continuous flow format. Our approach has high potential for the highly sensitive label-free detection of various proteins based on binding-induced conformation changes of immobilized DNA probes.
  • Language: English
  • Identifier: E-ISSN: 1520-6882 ; PMID: 21476599 Version:1 ; DOI: 10.1021/ac200236r

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