Assessment of the interaction between a synthetic epitope of troponin C and its specific antibody using a label-free faradaic impedimetric immunosensor and α-Keggin silicotungstic heteropolyacid as a redox probe

Mantzila, Aikaterini G.; Strongylis, Constantinos; Tsikaris, Vassilios; Prodromidis, Mamas I.

doi:10.1016/j.bios.2007.04.017

Cited by 18 publications

(15 citation statements)

References 31 publications

(33 reference statements)

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“…However, the backfilling did result in a current increase and a prominent decrease in the impedance signal (smaller semi-circles). A similar effect was observed in our previous studies [16] and in studies of other modified surfaces [42]. The impedance decrease upon backfilling can be explained by the easier charge transfer through tunneling across the shorter cysteine.…”

Section: Resultssupporting

confidence: 88%

Rapid Development of New Protein Biosensors Utilizing Peptides Obtained via Phage Display

Park

Dooley

et al. 2011

PLoS ONE

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There is a consistent demand for new biosensors for the detection of protein targets, and a systematic method for the rapid development of new sensors is needed. Here we present a platform where short unstructured peptides that bind to a desired target are selected using M13 phage display. The selected peptides are then chemically synthesized and immobilized on gold, allowing for detection of the target using electrochemical techniques such as electrochemical impedance spectroscopy (EIS). A quartz crystal microbalance (QCM) is also used as a diagnostic tool during biosensor development. We demonstrate the utility of this approach by creating a novel peptide-based electrochemical biosensor for the enzyme alanine aminotransferase (ALT), a well-known biomarker of hepatotoxicity. Biopanning of the M13 phage display library over immobilized ALT, led to the rapid identification of a new peptide (ALT5-8) with an amino acid sequence of WHWRNPDFWYLK. Phage particles expressing this peptide exhibited nanomolar affinity for immobilized ALT (Kd,app = 85±20 nM). The newly identified ALT5-8 peptide was then chemically synthesized with a C-terminal cysteine for gold immobilization. The performance of the gold-immobilized peptides was studied with cyclic voltammetry (CV), QCM, and EIS. Using QCM, the sensitivity for ALT detection was 8.9±0.9 Hz/(µg/mL) and the limit of detection (LOD) was 60 ng/mL. Using EIS measurements, the sensitivity was 142±12 impedance percentage change %/(µg/mL) and the LOD was 92 ng/mL. In both cases, the LOD was below the typical concentration of ALT in human blood. Although both QCM and EIS produced similar LODs, EIS is preferable due to a larger linear dynamic range. Using QCM, the immobilized peptide exhibited a nanomolar dissociation constant for ALT (Kd = 20.1±0.6 nM). These results demonstrate a simple and rapid platform for developing and assessing the performance of sensitive, peptide-based biosensors for new protein targets.

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Section: Resultssupporting

confidence: 88%

Rapid Development of New Protein Biosensors Utilizing Peptides Obtained via Phage Display

Park

Dooley

et al. 2011

PLoS ONE

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“…1 is especially suitable for electrochemical impedance based biosensors in which a semi-conductive biointerface is preferred to obtain a wide dynamic range and sensitivity [5,16]. In addition, EIS is an effective technique for the development of biosensors for real samples [17,18]. Atomic force microscopy (AFM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) techniques were used to investigate the immobilization of BmoLL on AuNpPANI to evaluate its interaction with the patients' sera infected by dengue serotypes 1, 2, and 3 (DEN1, DEN2, and DEN3).…”

Section: Introductionmentioning

confidence: 99%

Diagnosis of dengue infection using a modified gold electrode with hybrid organic–inorganic nanocomposite and Bauhinia monandra lectin

Andrade¹,

Oliveira²,

Melo³

et al. 2011

Journal of Colloid and Interface Science

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A sensitive and selective biosensor for dengue serotyping was successfully developed. The biosensor uses a novel gold nanoparticles-polyaniline hybrid composite (AuNpPANI) for the immobilization of Bauhinia monandra lectin (BmoLL). The nanocomposite was applied to a bare gold electrode surface by chemical adsorption, and BmoLL was subsequently electrostatically adsorbed to the nanocomposite-modified surface. Atomic force microscopy (AFM), cyclic voltammetry (CV) and electrochemical impedance (EI) techniques were applied to evaluate the immobilization of BmoLL on AuNpPANI. The AFM images for AuNpPANI-BmoLL-DEN systems indicate a homogenous, compact and dense film of the conjugate. In the EI analyses, an obvious difference of the electron transfer resistance between the AuNpPANI-modified electrode and the bare gold electrode was observed. Among three dengue serotypes studied, dengue serotype 2 (DEN2) has higher values for R(CT), and lower values for both n and Q. These are indications of a larger blocking effect and smaller capacitive dispersion, resulting from the higher agglutination of glycoproteins from the DEN2 sera. The selective BmoLL recognition for various dengue serotypes may be attributed to different patterns of glycoproteins in the sera produced by the glycoprotein immunoresponse from patients infected by the dengue virus.

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“…A serious disadvantage in the use of the above bulky redox molecules is the increase of the cost of the analysis, whereas in the case of the protein-assisted probing, sensors also suffer in terms of working simplicity, since extra compound(s) should be added in the measuring cell. Alternatively, bulky inorganic redox compounds, that is, silicotungstic acid hydrate (H 4 SiO 4 ·12WO 3 ·xH 2 O), have also been used as redox probes in faradic EIS experiments [59].…”

Section: Recent Developments In Immunosensorsmentioning

confidence: 99%

Impedimetric immunosensors—A review

Prodromidis

2010

Electrochimica Acta

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321

178

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Assessment of the interaction between a synthetic epitope of troponin C and its specific antibody using a label-free faradaic impedimetric immunosensor and α-Keggin silicotungstic heteropolyacid as a redox probe

Cited by 18 publications

References 31 publications

Rapid Development of New Protein Biosensors Utilizing Peptides Obtained via Phage Display

Rapid Development of New Protein Biosensors Utilizing Peptides Obtained via Phage Display

Diagnosis of dengue infection using a modified gold electrode with hybrid organic–inorganic nanocomposite and Bauhinia monandra lectin

Impedimetric immunosensors—A review

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