Electrochemical Aptamer-Based Sandwich Assays for the Detection of Explosives

Analytical Chemistry (Impact Factor: 5.83). 04/2012; 84(10):4245-7. DOI: 10.1021/ac300606n
Source: PubMed

ABSTRACT Electrochemical impedance spectroscopy (EIS) is used to detect 2,4,6-trinitrotoluene (TNT) in a novel sandwiched structure which relies on the specific interactions between (i) primary amine with TNT and (ii) TNT and anti-TNT aptamer. With pure targets, the assay has a sensitivity of 10(-14) M, a dynamic range of 10(-14)-10(-3) M, and employs a small sample volume (25 μL). The method's sensitivity is comparable to state of the art optical methods with the added advantages of electrochemical detection, which can be easily miniaturized and implemented into a hand-held device.

  • [Show abstract] [Hide abstract]
    ABSTRACT: Four custom-designed bidentate adsorbates having either ammonium or Boc-protected amino termini and either methanethiol or ethanethioate headgroups were prepared for the purpose of generating amine-terminated self-assembled monolayers (SAMs) on evaporated gold surfaces. These adsorbates utilize a phenyl-based framework to connect the headgroups to a single hexadecyloxy chain, extending the amino functionality away from the surface of gold, providing two regions within the adsorbate structure where intermolecular interactions contribute to the stability of the fully formed thin film. The structural features of the resulting SAMs were characterized by ellipsometry, X-ray photoelectron spectroscopy, and polarization modulation infrared reflection-absorption spectroscopy. The collected data were compared to those of eight additional SAMs formed from analogous monodentate alkanethiols and alkanethioacetates having either a similar aromatic framework or a simple alkyl chain connecting the headgroup to the tailgroup. Analysis of the data obtained for the full set of SAMs revealed that both the tailgroup and headgroup influenced the formation of a well-packed monolayer, with the Boc-protected amine-terminated alkanethiols producing films with superior surface bonding and adsorbate packing as compared to those formed with ammonium tailgroups or alkanethioacetate headgroups. Comparison of the structural differences before and after deprotection of the Boc-protected amine-terminated thiolate SAMs revealed that the bidentate adsorbate was the most resistant to desorption during the Boc-deprotection procedure. Furthermore, solution-phase thermal desorption tests performed to evaluate the thermal stability of the Boc-deprotected amine-terminated alkanethiolate films provided further evidence of the enhanced stability associated with SAMs formed from these bidentate adsorbates.
  • [Show abstract] [Hide abstract]
    ABSTRACT: A fluorometric sensing array was developed to detect and discern common military explosives and materials used in improvised explosive devices (IEDs). Xanthene-based fluorophores were chosen as reporter molecules and were exposed to explosive analytes in DMF solution. Unique responses were observed for each analyte, with the strongest observed responses resulting from trinitrobenzene (TNB) or trinitrotoluene (TNT) addition. Evidence for substantial electron transfer (ET) between TNB and TNT products and fluorophores was observed. Computational results indicate that anionic trinitroaromatic products are likely donating electrons to cationic fluorophores, leading to an observed enhancement of their emission.
    Analytical methods 01/2013; 5(20):5457. DOI:10.1039/c3ay40293b · 1.94 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Gold nanoparticle (AuNP) based colorimetric aptasensor have been developed for many analytes recently largely because of the ease of detection, high sensitivity, and potential for high-throughput analysis. Most of the target aptamers for detection have short sequences. However, the approach shows poor performance in terms of detection sensitivity for most of the long-sequence aptamers. To address this problem, for the first time, we split the 76 mer aptamer of 17β-estradiol into two short pieces to improve the AuNP based colorimetric sensitivity. Our results showed that the split P1 + P2 still retained the original 76 mer aptamer's affinity and specificity but increased the detection limit by 10-fold, demonstrating that as low as 0.1 ng/mL 17β-estradiol could be detected. The increased sensitivity may be caused by lower aptamer adsorption concentration and a lower affinity to the AuNPs of a short single-strand DNA (ssDNA) sequence. Our study provided a new way to use long-sequence aptamers to develop a highly sensitive AuNP-based colorimetric aptasensor.
    Scientific Reports 12/2014; 4:7571. DOI:10.1038/srep07571 · 5.08 Impact Factor