Using label-free screening technology to improve efficiency in drug discovery

Halai, Reena; Cooper, Matthew A.

doi:10.1517/17460441.2012.651121

Cited by 35 publications

(36 citation statements)

References 55 publications

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“…Thereby some of the caveats of traditional endpoint assays are circumvented: no labels (e.g. radioactive, fluorescent) or reporter genes are required and measurements can be performed continuously [4]. Cell types used include recombinant cell lines, cell lines endogenously expressing the target of interest and even patient-derived cells [5].…”

Section: Introductionmentioning

confidence: 99%

Constitutive activity of the metabotropic glutamate receptor 2 explored with a whole-cell label-free biosensor

Doornbos

Linden

Vereyken

et al. 2018

Biochemical Pharmacology

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Label-free cellular assays using a biosensor provide new opportunities for studying G protein-coupled receptor (GPCR) signaling. As opposed to conventional in vitro assays, integrated receptor-mediated cellular responses are determined in real-time rather than a single downstream signaling pathway. In this study, we examined the potential of a label-free whole cell impedance-based biosensor system (i.e. xCELLigence) to study the pharmacology of one GPCR in particular, the mGlu 2 receptor. This receptor is a target for the treatment of several psychiatric diseases such as schizophrenia and depression.After optimization of assay conditions to prevent interference of endogenous glutamate in the culture medium, detailed pharmacological assessments were performed. Concentration-response curves showed a concentration-dependent increase in impedance for agonists and positive allosteric modulators, whereas receptor inhibition by an antagonist or negative allosteric modulator resulted in a concentration-dependent decrease in cellular impedance. Interestingly, constitutive receptor activity was observed that was decreased by LY341495, which therefore behaved as an inverse agonist here, a property that was heretofore unappreciated. This was confirmed by concentration-dependent modulation of LY341495 potency and efficacy by a allosteric modulators.In summary, the use of the xCELLigence system to study mGlu 2 receptor pharmacology was validated. This is the first class C GPCR to be characterized extensively by such method, opening new avenues to study receptor pharmacology including inverse agonism and demonstrating its value for future drug discovery efforts of mGlu receptors as well as other GPCRs.

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Section: Introductionmentioning

confidence: 99%

Constitutive activity of the metabotropic glutamate receptor 2 explored with a whole-cell label-free biosensor

Doornbos

Linden

Vereyken

et al. 2018

Biochemical Pharmacology

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“…A theorem of linear systems states that the current response to the integrated form of a voltage signal is the integral form of the current response to the original voltage signal. 27 Therefore the integral of the Dirac delta function, the unit step function, is applied, and the current measured is the integral of the desired response, which can then be recovered by simple differentiation, after which the discrete time Fourier transform is performed to transform the data from the time domain to the frequency domain, (4) where f(n) is the nth observed signal, ω is the frequency, and ∆t is the sampling interval. The impedance is then calculated by dividing the voltage by the current.…”

Section: Ft-eismentioning

confidence: 99%

“…2 The impedance of cells can differ in response to the presence of toxins due to changes in cell morphology, disruption of the cell membrane, oxidative stress, etc., thus providing real-time, label-free, non-invasive toxin measurements. 3,4 In contrast, labels used in other cell-based sensing methods may interfere with normal cell functions and add preparation time and complexity to the biosensor. Furthermore, impedance can be measured continuously, whereas single point cell-based assays, e.g.…”

Section: Introductionmentioning

confidence: 99%

Nanofluidic structures for coupled sensing and remediation of toxins

Shaw

Contento

et al. 2014

SPIE Proceedings

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One foundational motivation for chemical sensing is that knowledge of the presence and level of a chemical agent informs decisions about treatment of the agent, for example by sequestration, separation or chemical conversion to a less harmful substance. Commonly the sensing and treatment steps are separate. However, the disjoint detection/treatment approach is neither optimal, nor required. Thus, we are investigating how nanostructured architectures can be constructed so that molecular transport (analyte/reagent delivery), chemical sensing (optical or electrochemical) and subsequent treatment can all be coupled in the same physical space during the same translocation event. Chemical sensors that are uniquely well-poised for integration into 3-D micro-/nanofluidic architectures include those based on plasmonics and impedance. Following detection, treatment can be substantially enhanced if mass transport limitations can be overcome. In this context, in situ generation of reactive species within confined geometries, such as nanopores or nanochannels, is of significant interest, because of its potential utility in overcoming mass transport limitations in chemical reactivity. Solvent electrolysis in electrochemically coupled nanochannels supporting electrokinetic flow for the generation of reactive species, can produce arbitrarily tunable quantities of reagents, such as O 2 or H 2 , in situ in close proximity to the site of a hydrogenation catalyst, for example. Semi-quantitative estimates of the local H 2 concentration are obtained by comparing the spatiotemporal fluorescence behavior and current measurements with finite element simulations accounting for electrolysis and subsequent convection and diffusion within the confined geometry. H 2 saturation can easily be achieved at modest overpotentials.

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“…Another class of label-free biosensors have been developed that focuses on morphological changes from a cellular systembased approach [22]. It has been known that changes to the cellular microenvironment can cause changes in cellular structure, behavior and viability [23].…”

Section: Current Label-free Techniques and Eismentioning

confidence: 99%

From Cellular Cultures to Cellular Spheroids: Is Impedance Spectroscopy a Viable Tool for Monitoring Multicellular Spheroid (MCS) Drug Models?

Alexander

Price

Bhansali

2013

IEEE Rev. Biomed. Eng.

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The use of 3-D multicellular spheroid (MCS) models is increasingly being accepted as a viable means to study cell-cell, cell-matrix and cell-drug interactions. Behavioral differences between traditional monolayer (2-D) cell cultures and more recent 3-D MCS confirm that 3-D MCS more closely model the in vivo environment. However, analyzing the effect of pharmaceutical agents on both monolayer cultures and MCS is very time intensive. This paper reviews the use of electrical impedance spectroscopy (EIS), a label-free whole cell assay technique, as a tool for automated screening of cell drug interactions in MCS models for biologically/physiologically relevant events over long periods of time. EIS calculates the impedance of a sample by applying an AC current through a range of frequencies and measuring the resulting voltage. This review will introduce techniques used in impedance-based analysis of 2-D systems; highlight recently developed impedance-based techniques for analyzing 3-D cell cultures; and discuss applications of 3-D culture impedance monitoring systems.

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Using label-free screening technology to improve efficiency in drug discovery

Cited by 35 publications

References 55 publications

Constitutive activity of the metabotropic glutamate receptor 2 explored with a whole-cell label-free biosensor

Constitutive activity of the metabotropic glutamate receptor 2 explored with a whole-cell label-free biosensor

Nanofluidic structures for coupled sensing and remediation of toxins

From Cellular Cultures to Cellular Spheroids: Is Impedance Spectroscopy a Viable Tool for Monitoring Multicellular Spheroid (MCS) Drug Models?

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