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Abstract In this work, we demonstrate the development of a rapid and label-free electrochemical biosensor to detectListeria monocytogenesusing a novel stimulus–response thiomer nanobrush material. Nanobrushes were developed via one-step simultaneous co-deposition of nanoplatinum (Pt) and alginate thiomers (ALG-thiomer). ALG-thiomer/Pt nanobrush platform significantly increased the average electroactive surface area of electrodes by 7 folds and maintained the actuation properties (pH-stimulated osmotic swelling) of the alginate. Dielectric behavior during brush actuation was characterized with positively, neutral, and negatively charged redox probes above and below the isoelectric point of alginate, indicating ALG-thiomer surface charge plays an important role in signal acquisition. The ALG-thiomer platform was biofunctionalized with an aptamer selective for the internalin A protein onListeriafor biosensing applications. Aptamer loading was optimized and various cell capture strategies were investigated (brush extended versus collapsed). Maximum cell capture occurs when the ALG-thiomer/aptamer is in the extended conformation (pH > 3.5), followed by impedance measurement in the collapsed conformation (pH < 3.5). Low concentrations of bacteria (5 CFU mL−1) were sensed from a complex food matrix (chicken broth) and selectivity testing against other Gram-positive bacteria (Staphylococcus aureus) indicate the aptamer affinity is maintained, even at these pH values. The new hybrid soft material is among the most efficient and fastest (17 min) forL. monocytogenesbiosensing to date, and does not require sample pretreatment, constituting a promising new material platform for sensing small molecules or cells.
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One-Step Fabrication of Stimuli-Responsive Chitosan-Platinum Brushes for Listeria monocytogenes Detection
Bacterial contamination in food-processing facilities is a critical issue that leads to outbreaks compromising the integrity of the food supply and public health. We developed a label-free and rapid electrochemical biosensor for Listeria monocytogenes detection using a new one-step simultaneous sonoelectrodeposition of platinum and chitosan (CHI/Pt) to create a biomimetic nanostructure that actuates under pH changes. The XPS analysis shows the effective co-deposition of chitosan and platinum on the electrode surface. This deposition was optimized to enhance the electroactive surface area by 11 times compared with a bare platinum–iridium electrode (p < 0.05). Electrochemical behavior during chitosan actuation (pH-stimulated osmotic swelling) was characterized with three different redox probes (positive, neutral, and negative charge) above and below the isoelectric point of chitosan. These results showed that using a negatively charged redox probe led to the highest electroactive surface area, corroborating previous studies of stimulus–response polymers on metal electrodes. Following this material characterization, CHI/Pt brushes were functionalized with aptamers selective for L. monocytogenes capture. These aptasensors were functional at concentrations up to 106 CFU/mL with no preconcentration nor extraneous reagent addition. Selectivity was assessed in the presence of other Gram-positive bacteria (Staphylococcus aureus) and with a food product (chicken broth). Actuation led to improved L. monocytogenes detection with a low limit of detection (33 CFU/10 mL in chicken broth). The aptasensor developed herein offers a simple fabrication procedure with only one-step deposition followed by functionalization and rapid L. monocytogenes detection, with 15 min bacteria capture and 2 min sensing.
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- Award ID(s):
- 1805512
- PAR ID:
- 10385431
- Date Published:
- Journal Name:
- Biosensors
- Volume:
- 11
- Issue:
- 12
- ISSN:
- 2079-6374
- Page Range / eLocation ID:
- 511
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Accepted Manuscript1.0
- Journal Article:
- https://doi.org/10.3390/bios11120511
