Electrochemical impedance spectroscopy biosensor for detection of active botulinum neurotoxin

Halliwell, Jennifer; Savage, Alison C.; Buckley, Nicholas; Gwenin, Christopher

doi:10.1016/j.sbsr.2014.08.002

Cited by 30 publications

(33 citation statements)

References 16 publications

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“… Voltammograms at a sweep rate of 50 mVs −1 in 0.1 M NaOH; the working electrodes were SNAP-25 modified Au (111) (─ ─ ─), VAMP modified Au (111) (●●●●) and a bare Au (111) slide (───). SNAP-25 modified Au (111) and bare Au (111) voltammograms reproduced from [ 25 ]. …”

Section: Resultsmentioning

confidence: 99%

“…The SNAP-25 modified electrodes were used to detect the activity of native botulinum neurotoxins BoNT/A, BoNT/C and BoNT/E. The use of this electrochemical impedance assay with the pharmaceutical Dysport ® containing BoNT/A has previously been reported [ 25 ]. The pharmaceutical sample contains stabilising excipients HSA and lactose, which could explain the lack of linear response observed using the pharmaceutical sample in the previously published work [ 25 ].…”

Section: Resultsmentioning

confidence: 99%

“…The use of this electrochemical impedance assay with the pharmaceutical Dysport ® containing BoNT/A has previously been reported [ 25 ]. The pharmaceutical sample contains stabilising excipients HSA and lactose, which could explain the lack of linear response observed using the pharmaceutical sample in the previously published work [ 25 ]. A typical Nyquist plot is shown in Figure 3 , and the charge transfer resistance was calculated by fitting the data to a Randles circuit.…”

Section: Resultsmentioning

confidence: 99%

“…Herein we report the development of two electrochemical impedance biosensors using the SNARE proteins SNAP-25 and VAMP on gold electrodes for the detection of different serotypes of botulinum neurotoxin. Previously we have reported the SNAP-25 electrochemical impedance biosensor for the detection of active BoNT/A in the pharmaceutical botulinum neurotoxin Dysport ® , capable of identifying active toxin in the presence of stabilising excipient human serum albumin (HSA) and lactose at concentrations as low as 25 fg/mL [ 24 , 25 ]. Building on from the success of this sensor for the detection of active BoNT/A in the pharmaceutical samples, we have applied it to the detection of native BoNT/A, as well as serotypes BoNT/C and BoNT/E.…”

Section: Introductionmentioning

confidence: 99%

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Botulinum Neurotoxin Serotypes Detected by Electrochemical Impedance Spectroscopy

Savage

Buckley

Halliwell

et al. 2015

Toxins

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Botulinum neurotoxin is one of the deadliest biological toxins known to mankind and is able to cause the debilitating disease botulism. The rapid detection of the different serotypes of botulinum neurotoxin is essential for both diagnosis of botulism and identifying the presence of toxin in potential cases of terrorism and food contamination. The modes of action of botulinum neurotoxins are well-established in literature and differ for each serotype. The toxins are known to specifically cleave portions of the SNARE proteins SNAP-25 or VAMP; an interaction that can be monitored by electrochemical impedance spectroscopy. This study presents a SNAP-25 and a VAMP biosensors for detecting the activity of five botulinum neurotoxin serotypes (A–E) using electrochemical impedance spectroscopy. The biosensors are able to detect concentrations of toxins as low as 25 fg/mL, in a short time-frame compared with the current standard methods of detection. Both biosensors show greater specificity for their compatible serotypes compared with incompatible serotypes and denatured toxins.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Botulinum Neurotoxin Serotypes Detected by Electrochemical Impedance Spectroscopy

Savage

Buckley

Halliwell

et al. 2015

Toxins

Self Cite

View full text Add to dashboard Cite

show abstract

“…The coupling of electrochemical transduction schemes has also been successfully demonstrated to generate immunosensors technologies that are highly selective and sensitive [18,[20][21][22]. Electrochemical biosensors can be divided by the electrical parameter that they measure (potentiometric, amperometric, voltammetric, capacitative, conductometric and impedance).…”

Section: Combination Of Electro-chemical and Optical Transductionsmentioning

confidence: 99%

Photonic tools for advanced sensing and imaging at the nanoscale.

Ghithan¹

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This dissertation reports a novel bio-sensing strategy based on single-mode, electro-active, integrated optical waveguide (SM-EA-IOW) platforms. It also reports the development of a super-resolved far-field optical imaging tool to enable optical, electronic, and spectroelectrochemical investigations at the nanoscale. SM-EA-IOW platforms with its outstanding sensitivity for spectroelectrochemical interrogation was combined with a sandwich bioassay for the development of a novel immunosensing based strategy for label-free detection of infectious pathogens. The strategy begins with the functionalization of the electroactive waveguide surface with a capturing antibody aimed at a specific target analyte. Once the target analyte is bound to the photonic interface, it promotes the binding of a secondary antibody that has been labeled with a redox active reporter. This labeled antibody reporter forms the analytical signal, which is linked uniquely to both the spectral and electrochemical properties of the redox probe designed to specifically recognize a target analyte. Based on this novel detection strategy experimental results in the interrogation of influenza A (H5N1) HA protein have reached an outstanding level of detection in the picomolar range. In vii addition, the novel label-free SM-EA-IOW bio-sensing strategy was successfully demonstrated for detection of gram-negative bacteria in present authentic clinical eye samples. Such demonstration has also shown the flexibility and ability of the new strategy to probe samples in in the microliter volume range, without any prior processing or pre-enrichment steps. As the groundwork towards the optimal operation of the novel sensing strategy, the effects of the adsorption process and the rate of electron transfer of redox bound species to the electrode surface were thoroughly studied. For each interface of a multilayer immunoassay assembly the surface density, the adsorption kinetic, and the electron-transfer rate of bound species of a redox-active protein were investigated using an optical impedance spectroscopy (OIS) technique based on measurements obtained with the SM-EA-IOW platform. Such methodology and acquired knowledge are crucial for the rational development of novel and advanced immuno-biosensors. Electrochemically modulated fluorescent molecules to be conjugated with relevant antibodies for creating an electroactive probe at the nanoscale was also investigated. Such capability has the potential to enable the development of an arrayed immunosensing technology. Fluorescence emission at the nanoscale suffers from two main restrictions, diffraction limit and photobleaching effects. To address these hinders, a modulated stimulated emission depletion microscope (STED) that is capable of achieving far-field super-resolved images was developed and used to reduce the power of the applied excitation and depletion laser beams diminish photobleaching effects in single-molecule sub-diffraction STED imaging. viii These two photonic devices provide new approaches for bio-sensi...

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