Multichannel Electrochemical Detection System for Quantitative Monitoring of PCR Amplification

Wojciechowski, Marek; Sundseth, R; Moreno, María Virginia; Henkens, Robert W.

doi:10.1093/clinchem/45.9.1690

Cited by 33 publications

(12 citation statements)

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“…Neutravidin (NAv) has the advantage over avidin or streptavidin of having a neutral isoelectric point (pI = 6.3) and thus to be neutral at pH 6-7 and to lead to the lowest nonspecific binding. 13 NAv can also be labelled by fluorophore groups, such as fluorescein (FITC), to follow the bioconjugation by optical spectroscopy. The two-site "sandwich-like" biotin/neutravidin pairing is widely exploited for immunoassays, especially ELISA bioassays 14,15,16 and appears as a linkage of choice for rapidly screening polymers' viability.…”

Section: 12mentioning

confidence: 99%

“…In this new step of our investigation, we describe the synthesis of a polylysine functionalized by the previously reported indium chelate [In( cb-te2pa )] + and a biotin (Btn) end group. The latter allows fast and efficient bioconjugation on beads or antibodies via the very strong and fast biotin–neutravidin noncovalent interactions. , Neutravidin (NAv) has the advantage over avidin or streptavidin of having a neutral isoelectric point (pI = 6.3) and thus to be neutral at pH 6–7 and to lead to the lowest nonspecific binding . NAv can also be labeled by fluorophore groups, such as fluorescein (FITC), to follow the bioconjugation by optical spectroscopy.…”

Section: Introductionmentioning

confidence: 99%

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Enabling Indium Channels for Mass Cytometry by Using Reinforced Cyclam-Based Chelating Polylysine

et al. 2020

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The synthesis of a polylysine polymer functionalized with the previously reported astonishingly inert [In(cb-te2pa)] + chelate was performed. A biotin end group allowed the conjugation to biotinylated beads by the intermediary of a Fluorescein isothiocyanate / neutravidin receptor. High quality Imaging Mass Cytometry TM trials, based on 115 In detection were performed to highlight the behavior of the material. Anti-CD20 antibody was labeled by the so obtained In(III) modified polylysine using the biotin/neutravidin interaction. Ramos (CD20[+]) and HL-60 (CD20[-]) cell lines were co-stained with the In(III) modified bioconjugate by finding the best staining conditions. Both immunofluorescence IF-M and Mass Cytometry analyses confirmed the specific binding of anti-CD20 onto Ramos cells. CyTOF R histograms constructed on the 115 In detection allowed to define and to separate, with a good signal-to-noise ratio, two populations (Ramos and HL-60). The inertness of In(III)-MCP-Nav over a three-month storage period was proved by performing new functionality tests involving Jurkat cells (CD20[-]) as well as the possibility of multiplexing trials involving the 115 In channel. The results assure a promising future use of the previously announced [In(cb-te2pa)] + complex based polymers for mass cytometry. Electronic Supplementary Information (ESI) available: [1H NMR of TFA-Lys-NCA, Poly(TFA-L-Lys)-Btn, Poly(L-Lys)-Btn and (PEGpoly(L-Lys)-Btn); GPC chromatogram of Poly(L-Lys)-Btn; NMR integration for PEG number evaluation;. Selected areas of blanks' and beads' 115 In integral intensities on IMC images of indium beads; Average 154 Sm counts per cell for Jurkat (CD45[+]) and Ramos (CD45[+]) cells; Overlayed histograms of Jurkat and Ramos cells].

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Section: 12mentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Enabling Indium Channels for Mass Cytometry by Using Reinforced Cyclam-Based Chelating Polylysine

et al. 2020

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“…Accordingly, the solid-contact ISEs can be used to simultaneously detect two molecules. Previous research has shown that an amplification factor of 10 4 can be obtained for the measurements involving a HRP label . The sensitivity could be improved further on the basis of signal amplification using multiple enzyme tags in a single binding event …”

Section: Resultsmentioning

confidence: 99%

Sequential and Selective Detection of Two Molecules with a Single Solid-Contact Chronopotentiometric Ion-Selective Electrode

Ding

Wang

et al. 2018

Anal. Chem.

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A polymeric membrane ion-selective electrode (ISE) is typically designed for the determination of one specific ion using a conventional method. In this work, we demonstrate a simple, versatile, and sensitive platform for simultaneous detection of two molecules with a single ISE. Under a series of periodic galvanostatic polarization, a solid-contact ISE without ion exchanger properties under zero-current conditions has been successfully used for simultaneous detection of two opposite charged ions with high sensitivity, good selectivity, and fast reversibility. By integration of biorecognition elements with the potentiometric measurement, highly sensitive and selective detection of a broad range of different molecular targets can be predicted. As a proof of concept, a potentiometric genosensor based on magnetic beads-enzyme sandwich assay has been designed for sensitive and selective detection of pathogenic bacteria Escherichia coli O157:H7 and Staphylococcus aureus. Under optimal conditions, two bacteria nucleic acid sequences can be detected simultaneously with high sensitivity and good selectivity by using a single solid-contact potentiometric ISE. The detection limits of Escherichia coli O157:H7 DNA and Staphylococcus aureus DNA are 120 and 54 fM (3σ), respectively. Because of its simplicity, this potentiometric technique based on ISE can be an attractive tool or detector to perform two analyte measurements.

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“…Screen-printed multielectrode sensor arrays are another technology that has enabled high-throughput electrochemical screening outside organic synthesis. These arrays were first designed to detect nucleic acids in multiple samples after polymerase chain reaction amplification and have since been applied to the detection of other biomolecules . The arrays generally consist of 96-well plates containing two carbon electrodes that have been screen-printed onto a printed circuit board.…”

Section: Recent History and State Of The Art Of Batch Electrochemical...mentioning

confidence: 99%

High-Throughput Electrochemistry: State of the Art, Challenges, and Perspective

Wills

Charvet

Battilocchio

et al. 2021

Org. Process Res. Dev.

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Electrochemical transformations involve complex parameter interactions, ranging from universal chemistry variables such as solvent and reagents to specialist factors including electrode material and current density. Hence, the development of a robust and scale-independent electrochemical reaction can currently be a challenge. High-throughput experimentation (HTE) is an enabling method for reaction optimization and robustness testing. Here we provide an industrial and academic perspective on the state of the art of the combination of HTE with electrochemical reaction optimization for applications, including scale-up. We then present our vision for a future in which HTE reduces barriers to wide adoption of electrochemistry across the field of chemical synthesis.

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Multichannel Electrochemical Detection System for Quantitative Monitoring of PCR Amplification

Cited by 33 publications

References 0 publications

Enabling Indium Channels for Mass Cytometry by Using Reinforced Cyclam-Based Chelating Polylysine

Enabling Indium Channels for Mass Cytometry by Using Reinforced Cyclam-Based Chelating Polylysine

Sequential and Selective Detection of Two Molecules with a Single Solid-Contact Chronopotentiometric Ion-Selective Electrode

High-Throughput Electrochemistry: State of the Art, Challenges, and Perspective

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