Rapid and Sensitive Inhibitor Screening Using Magnetically Modulated Biosensors

Roth, Shira; Danielli, Amos

doi:10.3390/s21144814

Cited by 7 publications

(5 citation statements)

References 42 publications

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“…The current techniques to identify protein–DNA interactions are usually time-consuming, expensive, or not sensitive enough. Recently, using an MMB platform, we demonstrated highly specific and sensitive detection of protein–protein interactions. , Here, we used the MMB platform to demonstrate, for the first time, a rapid and cost-effective approach to unambiguously detect protein–DNA interactions. The MMB-based protein–DNA assay was experimentally compared to EMSA, typically regarded as a gold standard for the detection of protein–DNA interactions.…”

Section: Discussionmentioning

confidence: 99%

“…Previously, we introduced a novel method, termed magnetic modulation biosensing (MMB), which can rapidly detect interacting proteins, , antibodies, and specific nucleic acid sequences. ,− While the principles of the MMB technology have been described before, − it has never been applied for detecting protein–DNA interactions. To detect protein–DNA interactions, two binding assays can be used.…”

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confidence: 99%

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Rapid Biosensing Method for Detecting Protein–DNA Interactions

et al. 2022

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Identifying and investigating protein−DNA interactions, which play significant roles in many biological processes, is essential for basic and clinical research. Current techniques for identification of protein−DNA interactions are laborious, timeconsuming, and suffer from nonspecific binding and limited sensitivity. To overcome these challenges and assess protein−DNA interactions, we use a magnetic modulation biosensing (MMB) system. In MMB, one of the interacting elements (protein or DNA) is immobilized to magnetic beads, and the other is coupled to a fluorescent molecule. Thus, the link between the magnetic bead and the fluorescent molecule is established only when binding occurs, enabling detection of the protein−DNA interaction. Using magnetic forces, the beads are concentrated and manipulated in a periodic motion in and out of a laser beam, producing a detectable oscillating signal. Using MMB, we detected protein−DNA interactions between short GC-rich DNA sequences and both a purified specificity protein 1 (Sp1) and an overexpressed Buttonhead (BTD) protein in a cell lysate. The specificity of the interactions was assessed using mutated DNA sequences and competition experiments. The assays were experimentally compared with commonly used electrophoretic mobility shift assay, which takes approximately 4−72 h. In comparison, the MMB-based assay's turnaround time is ∼2 h, and it provides unambiguous results and quantitative measures of performance. The MMB system uses simple and cheap components, making it an attractive alternative method over current costly and time-consuming techniques for analyzing protein−DNA interactions. Therefore, we anticipate that the MMB-based technique will significantly advance the detection of protein−DNA interactions in biomedical research.

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Section: Discussionmentioning

confidence: 99%

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Rapid Biosensing Method for Detecting Protein–DNA Interactions

et al. 2022

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“…Roth, S. et al developed a rapid and sensitive inhibitor screening tool using fluorescence and magnetic modulated biosensor (MMBS) which could screen small molecules that inhibit the interaction between S1 and ACE2. The assay based on MMBS has high sensitivity, minimal non-specific binding, and was much faster than the commonly used ELISA [ 107 ]. Han, Y.X.…”

Section: Drug Screening Sensors Based On Ace2mentioning

confidence: 99%

Research progress of biosensors for detection of SARS-CoV-2 variants based on ACE2

Wei

Zhang

et al. 2023

Talanta

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“…Furthermore, if agent 33 does indeed show activity against SARS-CoV-2 at concentrations of 1.56-2.60 µM [88,89]), there are no published data confirming the activity of substance 32 against SARS-CoV-2. In addition, in [90] authors used fluorescent and magnetically modulated biosensors to develop a rapid and sensitive tool for screening inhibitors of S-protein and ACE2 interactions and showed that even at the highest concentration of compound 32 in 100 µM, inhibition of interaction of RBD of SARS-CoV-2 S-protein and ACE2 does not occur. The authors of [95] carried out high-throughput screening (HTS) of an extensive library of drugs that can be repurposed to find potential therapeutic agents against SARS-CoV-2.…”

Section: Binding Site In the Receptor-binding Domain Of Smentioning

confidence: 99%

Molecular Modeling of Viral Type I Fusion Proteins: Inhibitors of Influenza Virus Hemagglutinin and the Spike Protein of Coronavirus

Borisevich

Zarubaev

Щербаков

et al. 2023

Viruses

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The fusion of viral and cell membranes is one of the basic processes in the life cycles of viruses. A number of enveloped viruses confer fusion of the viral envelope and the cell membrane using surface viral fusion proteins. Their conformational rearrangements lead to the unification of lipid bilayers of cell membranes and viral envelopes and the formation of fusion pores through which the viral genome enters the cytoplasm of the cell. A deep understanding of all the stages of conformational transitions preceding the fusion of viral and cell membranes is necessary for the development of specific inhibitors of viral reproduction. This review systematizes knowledge about the results of molecular modeling aimed at finding and explaining the mechanisms of antiviral activity of entry inhibitors. The first section of this review describes types of viral fusion proteins and is followed by a comparison of the structural features of class I fusion proteins, namely influenza virus hemagglutinin and the S-protein of the human coronavirus.

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Rapid and Sensitive Inhibitor Screening Using Magnetically Modulated Biosensors

Cited by 7 publications

References 42 publications

Rapid Biosensing Method for Detecting Protein–DNA Interactions

Rapid Biosensing Method for Detecting Protein–DNA Interactions

Research progress of biosensors for detection of SARS-CoV-2 variants based on ACE2

Molecular Modeling of Viral Type I Fusion Proteins: Inhibitors of Influenza Virus Hemagglutinin and the Spike Protein of Coronavirus

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