The Application of Whole Cell-Based Biosensors for Use in Environmental Analysis and in Medical Diagnostics

Gui, Qingyuan; Lawson, Tom; Shan, Suyan; Lü, Yan; Liu, Yong

doi:10.3390/s17071623

Cited by 285 publications

(156 citation statements)

References 105 publications

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“…In particular, electric cell-substrate impedance sensing (ECIS) has been effectively used to assess bioviability, demonstrating a potential for high-throughput monitoring of cells adhesion and proliferation in vitro. [4][5][6][7] However, these systems are generally not compatible with the microscopy techniques extensively used for life-sciences research, the current gold standard for cell physiology monitoring, because of either the opaque nature of the substrates and/or of the electrodes used for sensing. The excellent works by Róisín Owens and collaborators have in part overcome the limitations of standard ECIS, demonstrating organic electrochemical transistors able to detect and measure cell adhesion, proliferation and detachment in vitro with enhanced sensitivity and temporal resolution compared to standard technologies, with the added advantage of allowing for simultaneous electrical and optical analyses.…”

Section: Introductionmentioning

confidence: 99%

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Scuratti

Bonacchini

Bossio

et al. 2019

ACS Appl. Mater. Interfaces

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Cell-based biosensors constitute a fundamental tool in biotechnology, and their relevance has greatly increased in recent years as a result of a surging demand for reduced animal testing and for high-throughput and cost-effective in vitro screening platforms dedicated to environmental and biomedical diagnostics, drug development and toxicology. In this context, electrochemical/electronic cell-based biosensors represent a promising class of devices that enable long-term and real-time monitoring of cell physiology in a non-invasive and label-free fashion, with a remarkable potential for process automation and parallelization. Common limitations of this class of devices at large include the need for substrate surface modification strategies to ensure cell adhesion and immobilization, limited compatibility with complementary optical cell-probing techniques, and need for frequency-dependent measurements, which rely on elaborated equivalent electrical circuit models for data analysis and interpretation. We hereby demonstrate the monitoring of cell adhesion and detachment through the time-dependent variations in the quasi-static characteristic current curves of a highly stable electrolyte-gated transistor, based on an optically transparent network of printable polymer-wrapped semiconducting carbon-nanotubes. MAIN TEXTOptical cell viability assay and immunofluorescence staining: The proliferation was evaluated after 1, 2, 3, and 4 d in vitro. For each time point the medium was removed and replaced with RPMI without phenol red containing 0.5 mg mL −1 of MTT reagent (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, Sigma-Aldrich). Cells were reincubated at 37 °C for 3 h. Formazan salt produced by cells through reduction of MTT was then solubilized with 200 mL of ethanol and the absorbance was read at 560 and 690 nm (using a microplate reader TECAN Spark10M). The proliferation cell rate was calculated as the difference in absorbed intensity at 560 and 690 nm. Cells grown on glass coverslips coated with SWCNT networks were washed twice with PBS and fixed for 15 min at RT in 4 % paraformaldehyde and 4 % sucrose in 0.12 M sodium phosphate buffer, pH 7.4. Fixed cells were pre-incubated for 20 min in gelatin dilution buffer (GDB: 0.02 M sodium phosphate buffer, pH 7.4, 0.45 M NaCl, 0.2% (w/v) gelatin) containing 0.3% (v/v)

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Section: Introductionmentioning

confidence: 99%

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Scuratti

Bonacchini

Bossio

et al. 2019

ACS Appl. Mater. Interfaces

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“…12 Biosensors are typically designed by identifying natural biorecognition elements and then engineering them into a genetic circuit to drive the expression of a reporter gene in response to a desired target molecule. 12,13 Synthetic biosensors can be deployed as whole cells, in which the circuit is embedded in live bacteria, or cell-free biosensors, in which the circuit functions wholly within a cell extract or in vitro reaction mixture. Although theoretically simple, the engineering of either whole-cell or cell-free biosensors is unpredictable and often requires significant optimization.…”

Section: Introductionmentioning

confidence: 99%

Design of a transcriptional biosensor for the portable, on-demand detection of cyanuric acid

Li¹,

Silverman²,

Alam³

et al. 2019

Preprint

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Rapid molecular biosensing is an emerging application area for synthetic biology. Here, we engineer a portable biosensor for cyanuric acid (CYA), an analyte of interest for human and environmental health, using a LysR-type transcription regulator (LTTR) fromPseudomonas within the context of Escherichia coli gene expression machinery. To overcome cross-host portability challenges of LTTRs, we rationally engineered hybridPseudomonas-E. coli promoters by integrating DNA elements required for transcriptional activity and ligand-dependent regulation from both hosts, which enabled E. coli to function as whole-cell biosensor for CYA. To alleviate challenges of whole-cell biosensing, we adapted these promoter designs to function within a freeze-dried E. coli cell-free system to sense CYA. This portable, on-demand system robustly detects CYA within an hour from laboratory and real-world samples and works with both fluorescent and colorimetric reporters. This work elucidates general principles to facilitate the engineering of a wider array of LTTR-based environmental sensors.

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“…Accurate and sensitive detection of cancer cells or microorganisms is a significant issue in the clinical field, since it is directly related to human health. Consequently, rapid and sensitive detection can be helpful to prevent and control the potential diseases, with the suitable design of biosensors . Biosensors are, generally, the microelectronic systems or devices that make use of biological components to enable rapid, accurate and low‐level detection of various substances in body fluids or other media.…”

Section: Introductionmentioning

confidence: 99%

“…Consequently, rapid and sensitive detection can be helpful to prevent and control the potential diseases, with the suitable design of biosensors. [1] Biosensors are, generally, the microelectronic systems or devices that make use of biological components to enable rapid, accurate and low-level detection of various substances in body fluids or other media. The obvious virtue of using the biological components for detection lies in the highly specific property of biological molecules.…”

Section: Introductionmentioning

confidence: 99%

Single‐Cell Nanometric Coating Towards Whole‐Cell‐Based Biodevices and Biosensors

Dai

Wang

et al. 2018

ChemistrySelect

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Living cells with different coatings, like polyelectrolytes, biomolecules, metal nanoparticles or oxides, have more functions compared with the original state, which would be beneficial to many potential applications. Amongst these applications, whole‐cell biosensors/devices are commonly used for high‐throughput disease diagnosis, detection of toxicities, biomedical imaging, environmental monitoring or microbial fuel cells. As a key factor of the fabrication of whole cell biosensors/devices, the techniques for cell immobilization and cell viability have obvious influence on the sensitivity, stability and reproducibility. Cell coatings not only can be helpful to maintain the cell viability, but also can offer multifunction to facilitate the cell immobilization or orientation in the construction of biosensors/devices. Therefore, in this review, we are going to discuss various coating methods and materials, with the focus on their applications in the whole‐cell based biosensors and bio‐devices.

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The Application of Whole Cell-Based Biosensors for Use in Environmental Analysis and in Medical Diagnostics

Cited by 285 publications

References 105 publications

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Real-Time Monitoring of Cellular Cultures with Electrolyte-Gated Carbon Nanotube Transistors

Design of a transcriptional biosensor for the portable, on-demand detection of cyanuric acid

Single‐Cell Nanometric Coating Towards Whole‐Cell‐Based Biodevices and Biosensors

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