scholarly journals Development of an Optical Assay to Detect SARS-CoV-2 Spike Protein Binding Interactions with ACE2 and Disruption of these Interactions Using Electric Current

2020 ◽  
Author(s):  
Mahmoud Al Ahmad ◽  
Farah Mustafa ◽  
Neena Panicker ◽  
Tahir A. Rizvi
Keyword(s):  
Light Source ◽  
Protein Interactions ◽  
Viral Protein ◽  
Virus Transmission ◽  
Smart Phone ◽  
Spike Protein ◽  
Binding Potential ◽  
Label Free ◽  
Measured Intensity ◽  
Antigen Antibody

AbstractThis study proposes a novel optical method of detecting and reducing SARS-CoV-2 transmission, the virus responsible for the COVID-19 pandemic that is sweeping the world today. SARS-CoV-2 belongs to the β-coronaviruses characterized by the crown-shaped spike protein that protrudes out of the virus particles, giving the virus a “corona” shape; hence the name coronavirus. This virus is similar to the viruses that caused SARS (severe acute respiratory syndrome) and MERS (Middle East respiratory syndrome), the other two coronavirus epidemics that were recently contained within the last ten years. The technique being proposed uses a light source from a smart phone and a mobile spectrophotometer to enable detection of viral proteins in solution or paper as well as protein-protein interactions. The proof-of-concept is shown by detecting soluble preparations of spike protein subunits from SARS-CoV-2, followed by detection of the actual binding potential of the spike protein with its host receptor, the angiotensin-converting enzyme 2 (ACE2). The results are validated by showing that this method can detect antigen-antibody binding using two independent viral protein-antibody pairs. The binding could be detected optically both in solution and on a solid support such as nitrocellulose membrane. Finally, this technique is combined with DC bias to show that introduction of a current into the system can be used to disrupt the antigen-antibody reaction, suggesting that the proposed extended technique can be a potential means of not only detecting the virus, but also reducing virus transmission by disrupting virus-receptor interactions electrically.SignificanceThe measured intensity of light can reveal information about different cellular parameters under study. When light passes through a bio-composition, the intensity is associated with its content. The nuclei size, cell shape and the refractive index variation of cells contributes to light intensity. In this work, an optical label-free real time detection method incorporating the smartphone light source and a portable mini spectrometer for SARS-CoV-2 detection was developed based on the ability of its spike protein to interact with the ACE2 receptor. The light interactions with control and viral protein solutions were capable of providing a quick decision regarding whether the sample under test was positive or negative, thus enabling SARS-CoV-2 detection in a rapid manner.

scholarly journals Optical Detection of SARS-CoV-2 Utilizing Antigen-Antibody Binding Interactions

Sensors ◽  
2021 ◽  
Vol 21 (19) ◽  
pp. 6596
Author(s):  
Mahmoud Al Ahmad ◽  
Farah Mustafa ◽  
Neena Panicker ◽  
Tahir A. Rizvi
Keyword(s):  
Optical Detection ◽  
Smart Phone ◽  
Antibody Binding ◽  
Binding Potential ◽  
Proof Of Concept ◽  
Binding Interactions ◽  
Protein Subunits ◽  
Optical Responses ◽  
Analysis Technique ◽  
Antigen Antibody

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for the coronavirus disease (COVID-19) pandemic, is sweeping the world today. This study investigates the optical detection of SARS-CoV-2, utilizing the antigen-antibody binding interactions utilizing a light source from a smart phone and a portable spectrophotometer. The proof-of-concept is shown by detecting soluble preparations of spike protein subunits from SARS-CoV-2, followed by detection of the actual binding potential of the SARS-CoV-2 proteins with their corresponding antigens. The measured binding interactions for RBD and NCP proteins with their corresponding antibodies under different conditions have been measured and analyzed. Based on these observations, a “hump or spike” in light intensity is observed when a specific molecular interaction takes place between two proteins. The optical responses could further be analyzed using the principle component analysis technique to enhance and allows precise detection of the specific target in a multi-protein mixture.

scholarly journals Analysis of Zika virus capsid-Aedes aegypti mosquito interactome reveals pro-viral host factors critical for establishing infection

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Rommel J. Gestuveo ◽  
Jamie Royle ◽  
Claire L. Donald ◽  
Douglas J. Lamont ◽  
Edward C. Hutchinson ◽  
...  
Keyword(s):  
Aedes Aegypti ◽  
Protein Interactions ◽  
Zika Virus ◽  
Label Free ◽  
Protein Protein Interactions ◽  
Zikv Infection ◽  
Ubiquitin Protein Ligase ◽  
Mosquito Cells ◽  
Endoplasmic Reticulum Protein ◽  
Arboviral Diseases

AbstractThe escalating global prevalence of arboviral diseases emphasizes the need to improve our understanding of their biology. Research in this area has been hindered by the lack of molecular tools for studying virus-mosquito interactions. Here, we develop an Aedes aegypti cell line which stably expresses Zika virus (ZIKV) capsid proteins in order to study virus-vector protein-protein interactions through quantitative label-free proteomics. We identify 157 interactors and show that eight have potentially pro-viral activity during ZIKV infection in mosquito cells. Notably, silencing of transitional endoplasmic reticulum protein TER94 prevents ZIKV capsid degradation and significantly reduces viral replication. Similar results are observed if the TER94 ortholog (VCP) functioning is blocked with inhibitors in human cells. In addition, we show that an E3 ubiquitin-protein ligase, UBR5, mediates the interaction between TER94 and ZIKV capsid. Our study demonstrates a pro-viral function for TER94/VCP during ZIKV infection that is conserved between human and mosquito cells.

scholarly journals Nanoparticle Determination in Water by LED-Excited Surface Plasmon Resonance Imaging

Chemosensors ◽  
2021 ◽  
Vol 9 (7) ◽  
pp. 175
Author(s):  
Lukas Wunderlich ◽  
Peter Hausler ◽  
Susanne Märkl ◽  
Rudolf Bierl ◽  
Thomas Hirsch
Keyword(s):  
Light Source ◽  
Self Assembled Monolayers ◽  
Receptor Interaction ◽  
Complex Nature ◽  
Label Free ◽  
Quality Of Water ◽  
Spr Imaging ◽  
Online Sensor ◽  
Assembled Monolayers

The increasing popularity of nanoparticles in many applications has led to the fact that these persistent materials pollute our environment and threaten our health. An online sensor system for monitoring the presence of nanoparticles in fresh water would be highly desired. We propose a label-free sensor based on SPR imaging. The sensitivity was enhanced by a factor of about 100 by improving the detector by using a high-resolution camera. This revealed that the light source also needed to be improved by using LED excitation instead of a laser light source. As a receptor, different self-assembled monolayers have been screened. It can be seen that the nanoparticle receptor interaction is of a complex nature. The best system when taking sensitivity as well as reversibility into account is given by a dodecanethiol monolayer on the gold sensor surface. Lanthanide-doped nanoparticles, 29 nm in diameter and with a similar refractive index to the most common silica nanoparticles were detected in water down to 1.5 µg mL−1. The sensor can be fully regenerated within one hour without the need for any washing buffer. This sensing concept is expected to be easily adapted for the detection of nanoparticles of different size, shape, and composition, and upon miniaturization, suitable for long-term applications to monitor the quality of water.

scholarly journals Affinity Sensors for the Diagnosis of COVID-19

Micromachines ◽  
2021 ◽  
Vol 12 (4) ◽  
pp. 390
Author(s):  
Maryia Drobysh ◽  
Almira Ramanaviciene ◽  
Roman Viter ◽  
Arunas Ramanavicius
Keyword(s):  
Field Effect Transistors ◽  
Resonance Field ◽  
Analytical Signal ◽  
Nucleic Acid Hybridization ◽  
Detection Methods ◽  
Label Free ◽  
Infected People ◽  
Label Free Detection ◽  
Main Instrument ◽  
Antigen Antibody

The coronavirus disease 2019 (COVID-19) outbreak caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) was proclaimed a global pandemic in March 2020. Reducing the dissemination rate, in particular by tracking the infected people and their contacts, is the main instrument against infection spreading. Therefore, the creation and implementation of fast, reliable and responsive methods suitable for the diagnosis of COVID-19 are required. These needs can be fulfilled using affinity sensors, which differ in applied detection methods and markers that are generating analytical signals. Recently, nucleic acid hybridization, antigen-antibody interaction, and change of reactive oxygen species (ROS) level are mostly used for the generation of analytical signals, which can be accurately measured by electrochemical, optical, surface plasmon resonance, field-effect transistors, and some other methods and transducers. Electrochemical biosensors are the most consistent with the general trend towards, acceleration, and simplification of the bioanalytical process. These biosensors mostly are based on the determination of antigen-antibody interaction and are robust, sensitive, accurate, and sometimes enable label-free detection of an analyte. Along with the specification of biosensors, we also provide a brief overview of generally used testing techniques, and the description of the structure, life cycle and immune host response to SARS-CoV-2, and some deeper details of analytical signal detection principles.

scholarly journals Therapeutic development by repurposing drugs targeting SARS-CoV-2 spike protein interactions by simulation studies

2021 ◽  
Author(s):  
Qazi Mohammad Sajid Jamal ◽  
Varish Ahmad ◽  
Ali H Alharbi ◽  
Mohammad Azam Ansari ◽  
Mohammad A Alzohairy ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Spike Protein ◽  
Simulation Studies ◽  
Therapeutic Development

scholarly journals Proteomic Discovery of VEEV E2-Host Partner Interactions Identifies GRP78 Inhibitor HA15 as a Potential Therapeutic for Alphavirus Infections

Pathogens ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 283
Author(s):  
Michael D. Barrera ◽  
Victoria Callahan ◽  
Ivan Akhrymuk ◽  
Nishank Bhalla ◽  
Weidong Zhou ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Protein Trafficking ◽  
Viral Protein ◽  
Sindbis Virus ◽  
Encephalitis Virus ◽  
Host Proteins ◽  
Atp Production ◽  
Equine Encephalitis Virus ◽  
E2 Glycoprotein ◽  
Late Stages

Alphaviruses are a genus of the Togaviridae family and are widely distributed across the globe. Venezuelan equine encephalitis virus (VEEV) and eastern equine encephalitis virus (EEEV), cause encephalitis and neurological sequelae while chikungunya virus (CHIKV) and Sindbis virus (SINV) cause arthralgia. There are currently no approved therapeutics or vaccines available for alphaviruses. In order to identify novel therapeutics, a V5 epitope tag was inserted into the N-terminus of the VEEV E2 glycoprotein and used to identify host-viral protein interactions. Host proteins involved in protein folding, metabolism/ATP production, translation, cytoskeleton, complement, vesicle transport and ubiquitination were identified as VEEV E2 interactors. Multiple inhibitors targeting these host proteins were tested to determine their effect on VEEV replication. The compound HA15, a GRP78 inhibitor, was found to be an effective inhibitor of VEEV, EEEV, CHIKV, and SINV. VEEV E2 interaction with GRP78 was confirmed through coimmunoprecipitation and colocalization experiments. Mechanism of action studies found that HA15 does not affect viral RNA replication but instead affects late stages of the viral life cycle, which is consistent with GRP78 promoting viral assembly or viral protein trafficking.

scholarly journals Biosensors for the Determination of SARS-CoV-2 Virus and Diagnosis of COVID-19 Infection

2022 ◽  
Vol 23 (2) ◽  
pp. 666
Author(s):  
Maryia Drobysh ◽  
Almira Ramanaviciene ◽  
Roman Viter ◽  
Chien-Fu Chen ◽  
Urte Samukaite-Bubniene ◽  
...  
Keyword(s):  
Current Trend ◽  
Analytical Signal ◽  
Diagnostic Methods ◽  
Detection Methods ◽  
Label Free ◽  
Free Mode ◽  
Rna Hybridisation ◽  
Antigen Antibody ◽  
Affinity Interaction

Monitoring and tracking infection is required in order to reduce the spread of the coronavirus disease 2019 (COVID-19), induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). To achieve this goal, the development and deployment of quick, accurate, and sensitive diagnostic methods are necessary. The determination of the SARS-CoV-2 virus is performed by biosensing devices, which vary according to detection methods and the biomarkers which are inducing/providing an analytical signal. RNA hybridisation, antigen-antibody affinity interaction, and a variety of other biological reactions are commonly used to generate analytical signals that can be precisely detected using electrochemical, electrochemiluminescence, optical, and other methodologies and transducers. Electrochemical biosensors, in particular, correspond to the current trend of bioanalytical process acceleration and simplification. Immunosensors are based on the determination of antigen-antibody interaction, which on some occasions can be determined in a label-free mode with sufficient sensitivity.

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