A new real-time method for investigation of affinity properties and binding kinetics of magnetic nanoparticles

Орлов, А. В.; Nikitin, Maxim P.; Bragina, V.A.; Znoyko, S.L.; Zaikina, Marina N.; Ksenevich, Tatiana I.; Gorshkov, B. G.; Никитин, П. И.

doi:10.1016/j.jmmm.2014.10.019

Cited by 41 publications

(19 citation statements)

References 42 publications

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“…For development of the sensitive and rapid assays, we carried out selection of optimal antibodies and conjugates of fluorescein with BSA by their binding kinetic characterization using modified label-free biosensors based on spectral-phase interferometry (SPI) [13] and spectral-correlation interferometry (SCI) [14] that provide real-time monitoring of thickness of a layer of biomolecular complexes bound on the sensor chip surface due to biochemical reactions. The techniques use microscopic cover glass slips as two-beam reflective interferometers and affordable sensor chips simultaneously.…”

Section: Label-free Monitoring Of Molecular Bindingsmentioning

confidence: 99%

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Volumetric registration of magnetic nanoparticles for optimization of quantitative immunochromatographic assays for detection of small molecules

et al. 2018

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Precise quantitative and highly sensitive detection of small molecules (haptens) is highly demanded in medicine, food quality control, in vitro diagnostics, criminalistics, environmental monitoring, etc. In the present work, the magnetic method of particle quantification and the optical methods of spectral correlation and spectral phase interferometry complement each other for optimization of a quantitative assay for measuring concentrations of small molecules. The assay employs magnetic nanoparticles as labels in rapid immunochromatographic format. The approach was demonstrated with fluorescein as a model molecule. The interferometric label-free biosensors were employed for selection of optimal reagents that produced high specificity and sensitivity. The method of magnetic particle quantification counted the magnetic labels over the entire volume of the immunochromatographic membrane to provide their distribution along the test strip. Such distribution was used for optimization of such parameters as concentrations of the used reagents and of antibody immobilized on the labels, amount of the labels and conjugates of haptens with protein carriers to realize the advanced quantitative immunochromatographic assay.

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Section: Label-free Monitoring Of Molecular Bindingsmentioning

confidence: 99%

“…The techniques use microscopic cover glass slips as two-beam reflective interferometers and affordable sensor chips simultaneously. These biosensors showed high efficiency for optimization of Ab immobilization interfaces and development of protocols for sandwich magnetic immunoassays [14].…”

Section: Label-free Monitoring Of Molecular Bindingsmentioning

confidence: 99%

Volumetric registration of magnetic nanoparticles for optimization of quantitative immunochromatographic assays for detection of small molecules

et al. 2018

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“…It should be noted that the SCI method demonstrated here for label-free immunosensing can also be employed as an effective auxiliary tool for development of various assays including those based on fluorescent, magnetic [22,23], or plasmonic labels on glass surface. Wide employment of the glass substrates for fluorescent microarrays and biochips is due to low-level autofluorescence.…”

Section: B-chipsmentioning

confidence: 99%

“…The simplest way to improve LOD in the existing biosensors is additional amplification steps with non-labeled tracer antibody for sandwich assays [20] or with various nanoparticles [10,11,[21][22][23]. Although these steps diminish the beauty of Breagentless^assays; however, we believe that such biosensors may be attractive because of much shorter assay time as compared with ELISA and due to their compatibility for multi-analyte quantitative assays in contract to current lateral flow tests [13].…”

Section: Introductionmentioning

confidence: 99%

Direct immunosensing by spectral correlation interferometry: assay characteristics versus antibody immobilization chemistry

et al. 2015

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A 3-channel biosensor based on spectral correlation interferometry (SCI) has been adapted for direct optical detection of antigens by measuring changes in thickness of a biolayer on functionalized glass slips employed as affordable single-use sensor chips. The instrument is insensitive to the bulk refractive index of a solution under test and provides signals in metrological units (pm or nm). Using real-time monitoring with the SCI, protocols for fabrication of sensor chips with different functional (epoxylated, carboxylated, and biotinylated) surfaces for antibody immobilization have been developed and optimized to minimize chip-to-chip variations and achieve better limit of detection (LOD), shorter assay time, and longer shelf life. The optimized coupling surfaces have been compared for detection of human serum albumin (HSA) used as a model agent of medical significance. The dynamic ranges for measuring the HSA concentration were 0.07-20, 0.12-30, and 0.25-10 μg/ml, and the assay durations were less than 20, 15, and 30 min for the epoxylated, carboxylated, and biotinylated chips, respectively. The advantages of each type of sensor chip have been shown, namely, the carboxylated chips feature the shortest assay time, the epoxylated ones demonstrate the best LOD, and the biotinylated chips exhibit the longest shelf life in an unprotected environment. The developed protocols of antibody immobilization can be used in different biosensors and assay techniques including those based on fluorescent, magnetic or plasmonic labels, etc. The SCI is well compatible with various partially transparent layers used in biosensing and with microarrays for multi-analyte detection.

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“…One of the research fields that may greatly benefit from these capabilities is development of the advanced nanoagents for targeted drug delivery . Indeed, investigation of the particle–cell interactions with IFC can yield statistical information about the agent specificity , kinetics of its internalization and subsequent localization within the cells , expression levels of cells surface receptors , and their affinity. Furthermore, it can quantitatively assess the cytotoxicity of the agent and its payload .…”

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confidence: 99%

Precise Quantitative Analysis of Cell Targeting by Particle‐Based Agents Using Imaging Flow Cytometry and Convolutional Neural Network

et al. 2019

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Understanding the intricacies of particle–cell interactions is essential for many applications such as imaging, phototherapy, and drug/gene delivery, because it is the key to accurate control of the particle properties for the improvement of their therapeutic and diagnostic efficiency. Recently, high‐throughput methods have emerged for the detailed investigation of these interactions. For example, imaging flow cytometry (IFC) collects up to 60,000 images of cells per second (in 12 optical channels) and provides information about morphology and organelle localization in combination with fluorescence and side scatter intensity data. However, analysis of IFC data is extremely difficult to perform using conventional methods that calculate integral parameters or use mask‐based object recognition. Here, we show application of a convolutional neural network (CNN) for precise quantitative analysis of particle targeting of cells using IFC data. CNN provides high‐throughput object detection with almost human precision but avoids the subjective choice of image processing parameters that often leads to incorrect data interpretation. The method allows accurate counting of cell‐bound particles with reliable discrimination from the nonbound particles in the field of view. The proposed method expands capabilities of spot counting applications (such as organelle counting, quantification of cell–cell and cell–bacteria interactions) and is going to be useful not only for high‐throughput analysis of IFC data but also for other imaging techniques. © 2019 International Society for Advancement of Cytometry

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A new real-time method for investigation of affinity properties and binding kinetics of magnetic nanoparticles

Cited by 41 publications

References 42 publications

Volumetric registration of magnetic nanoparticles for optimization of quantitative immunochromatographic assays for detection of small molecules

Volumetric registration of magnetic nanoparticles for optimization of quantitative immunochromatographic assays for detection of small molecules

Direct immunosensing by spectral correlation interferometry: assay characteristics versus antibody immobilization chemistry

Precise Quantitative Analysis of Cell Targeting by Particle‐Based Agents Using Imaging Flow Cytometry and Convolutional Neural Network

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