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1.
Talanta ; 246: 123429, 2022 Aug 15.
Article in English | MEDLINE | ID: covidwho-1768562

ABSTRACT

Viral infection has been one of the major health issues for human life. The real-time reverse transcription polymerase chain reaction (RT-PCR)-based detection has primarily been used for virus detection as a highly reliable procedure. However, it is a relatively long and multi-stage process. In addition, required skilled personnel and complex instrumentation presents difficulties in large scale monitoring efforts. Therefore, we report here a direct and fast detection method for CoV-2 genome as applied in the nose-throat swab samples without any further processing. The detection principle is based on fluorescein-loaded mesoporous silica nanoparticles capped by specific gene sequences probes immobilized on the surface of the nanoparticles. Upon hybridization with the target viral genome, the fluorescein molecules were released from the mesopores. Testing with synthetic oligonucleotides, the NSP12 gene-based detection resulted in a strong signal. Target detection time could be optimized to 15 min and the limit of detection was 1.4 RFU with 84% sensitivity with clinical samples (n = 43).


Subject(s)
COVID-19 , Nanoparticles , Fluoresceins , Humans , RNA, Viral/genetics , SARS-CoV-2 , Sensitivity and Specificity , Silicon Dioxide
2.
Int J Environ Res Public Health ; 19(6)2022 03 18.
Article in English | MEDLINE | ID: covidwho-1760602

ABSTRACT

Masks are effective for preventing the spread of COVID-19 and other respiratory infections. If antimicrobial properties can be applied to the non-woven fabric filters in masks, they can become a more effective countermeasure against human-to-human and environmental infections. We investigated the possibilities of carrying antimicrobial agents on the fiber surfaces of non-woven fabric filters by applying silica-resin coating technology, which can form silica-resin layers on such fabrics at normal temperature and pressure. Scanning electron microscopy and electron probe microanalysis showed that a silica-resin layer was formed on the fiber surface of non-woven fabric filters. Bioassays for coronavirus and quantitative reverse transcription-polymerase chain reactions (RT-PCR) revealed that all antimicrobial agents tested loaded successfully onto non-woven fabric filters without losing their inactivation effects against the human coronavirus (inhibition efficacy: >99.999%). These results indicate that this technology could be used to load a functional substance onto a non-woven fabric filter by vitrifying its surface. Silica-resin coating technology also has the potential of becoming an important breakthrough not only in the prevention of infection but also in various fields, such as prevention of building aging, protection of various cultural properties, the realization of a plastic-free society, and prevention of environmental pollution.


Subject(s)
COVID-19 , Silicon Dioxide , Antiviral Agents , COVID-19/prevention & control , Humans , Masks , Textiles
3.
IET Nanobiotechnol ; 16(3): 85-91, 2022 May.
Article in English | MEDLINE | ID: covidwho-1758388

ABSTRACT

Mesoporous magnetic nanoparticles of haematite were synthesised using plant extracts according to bioethics principles. The structural, physical and chemical properties of mesoporous Fe2 O3 nanoparticles synthesised with the green chemistry approach were evaluated by XRD, SEM, EDAX, BET, VSM and HRTEM analysis. Then, their toxicity against normal HUVECs and MCF7 cancer cells was evaluated by MTT assay for 48 h. These biogenic mesoporous magnetic nanoparticles have over 71% of doxorubicin loading efficiency, resulting in a 50% reduction of cancer cells at a 0.5 µg.ml-1 concentration. Therefore, it is suggested that mesoporous magnetic nanoparticles be used as a multifunctional agent in medicine (therapeutic-diagnostic). The produced mesoporous magnetic nanoparticles with its inherent structural properties such as polygonal structure (increasing surface area to particle volume) and porosity with large pore volume became a suitable substrate for loading the anti-cancer drug doxorubicin.


Subject(s)
Nanoparticles , Silicon Dioxide , Doxorubicin/chemistry , Doxorubicin/pharmacology , Drug Carriers/chemistry , Drug Delivery Systems/methods , Drug Liberation , Humans , Nanoparticles/chemistry , Porosity , Silicon Dioxide/chemistry
4.
Colloids Surf B Biointerfaces ; 213: 112400, 2022 May.
Article in English | MEDLINE | ID: covidwho-1670367

ABSTRACT

The influence of the properties of different solid substrates on the tethering of two antibodies, IgG1-CR3022 and IgG1-S309, which were specifically engineered for the detection of SARS-CoV-2, has been examined at the molecular level using conventional and accelerated Molecular Dynamics (cMD and aMD, respectively). Two surfaces with very different properties and widely used in immunosensors for diagnosis, amorphous silica and the most stable facet of the face-centered cubic gold structure, have been considered. The effects of such surfaces on the structure and orientation of the immobilized antibodies have been determined by quantifying the tilt and hinge angles that describe the orientation and shape of the antibody, respectively, and the dihedrals that measure the relative position of the antibody arms with respect to the surface. Results show that the interactions with amorphous silica, which are mainly electrostatic due to the charged nature of the surface, help to preserve the orientation and structure of the antibodies, especially of the IgG1-CR3022, indicating that the primary sequence of those antibodies also plays some role. Instead, short-range van der Waals interactions with the inert gold surface cause a higher degree tilting and fraying of the antibodies with respect to amorphous silica. The interactions between the antibodies and the surface also affect the correlation among the different angles and dihedrals, which increases with their strength. Overall, results explain why amorphous silica substrates are frequently used to immobilize antibodies in immunosensors.


Subject(s)
Biosensing Techniques , COVID-19 , Antibodies, Monoclonal , Antibodies, Neutralizing/chemistry , Biosensing Techniques/methods , COVID-19/diagnosis , Gold/chemistry , Humans , Immunoassay/methods , Immunoglobulin G , SARS-CoV-2 , Silicon Dioxide
5.
ChemistryOpen ; 10(12): 1244-1250, 2021 12.
Article in English | MEDLINE | ID: covidwho-1598867

ABSTRACT

Rice husk, one of the main side products in the rice production, and its sustainable management represent a challenge in many countries. Herein, we describe the use of this abundant agricultural bio-waste as feedstock for the preparation of silver-containing carbon/silica nano composites with antimicrobial properties. The synthesis was performed using a fast and cheap methodology consisting of wet impregnation followed by pyrolysis, yielding C/SiO2 composite materials doped with varying amounts of silver from 28 to 0.001 wt %. The materials were fully characterized and their antimicrobial activity against ESKAPE pathogens, namely E. faecium, S. aureus, K. pneumoniae, A. baumannii, P. aeruginosa, and E. coli, and the pathogenic yeast C. albicans was investigated. Sensitivities of these strains against the prepared materials were demonstrated, even with exceptional low amounts of 0.015 m% silver. Hence, we report a straightforward method for the synthesis of antimicrobial agents from abundant sources which addresses urgent questions like bio-waste valorization and affordable alternatives to increasingly fewer effective antibiotics.


Subject(s)
Anti-Infective Agents , Oryza , Anti-Bacterial Agents/pharmacology , Anti-Infective Agents/pharmacology , Carbon , Escherichia coli , Silicon Dioxide , Silver/pharmacology , Staphylococcus aureus
6.
Talanta ; 240: 123211, 2022 Apr 01.
Article in English | MEDLINE | ID: covidwho-1595920

ABSTRACT

A label-free electrochemical biosensing approach as an appropriate analysis technique for SARS-CoV-2 spike protein (SARS-CoV-2 S-protein) was investigated to facilitate the diagnosis of coronavirus in real samples. It is crucial to construct diagnostic features that can rapidly identify infected individuals to limit the spread of the virus and assign treatment choices. Therefore, a novel and selective method using SiO2@UiO-66 and a label-free electrochemical immunoassay for rapidly detecting spike protein. The development of innovative approaches for direct viral detection employing simplified and ideally reagent-free assays is a pressing and difficult topic. The absence of speedy and effective ways to diagnose viral diseases especially SARS-CoV-2 on demand has worsened the issue of combating the COVID-19 pandemic. The developed electrode illustrated a wide dynamic range of 100.0 fg mL-1 to 10.0 ng mL-1 with low limit detection. Therefore, the as-fabricated electrochemical SARS-CoV-2 S-protein sensor suggests an appropriate perspective in the point-of-care system, within 5.0 min, in nasal samples with satisfactory recovery.


Subject(s)
Biosensing Techniques , COVID-19 , Electrochemical Techniques , Humans , Immunoassay , Metal-Organic Frameworks , Pandemics , Phthalic Acids , SARS-CoV-2 , Silicon Dioxide , Spike Glycoprotein, Coronavirus
7.
Zhong Nan Da Xue Xue Bao Yi Xue Ban ; 46(11): 1203-1211, 2021 Nov 28.
Article in English, Chinese | MEDLINE | ID: covidwho-1575959

ABSTRACT

OBJECTIVES: Coronavirus disease 2019 (COVID-19) is an acute respiratory infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). SARS-CoV-2 can damage the myocardium directly, or activate the immune system, trigger a cytokine storm, and cause inflammatory cells to infiltrate the myocardial tissue and damage the myocardium. This study is based on the sequencing data to analyze the changes in gene expression of cardiomyocytes and macrophages after SARS-CoV-2 infection, and explore the potential effects of SARS-CoV-2 on the heart and immune system. METHODS: The public data set GSE151879 was retrieved. The online software Network Analyst was used to preprocess the data, and the differentially expressed genes (DEGs) [log2(fold change)>2, adjusted P-value<0.05] screening between the infection group and the control group in cardiomyocytes, human embryonic stem cell-derived cardiomyocytes, and macrophages were screened. Consistent common differentially expressed genes (CCDEGs) with the same expression pattern in cardiomyocytes and macrophages were obtained, and the online analysis software String was used to conduct enrichment analysis of their biological functions and signal pathways. Protein-protein interaction network, transcription factor-gene interaction network, miRNA-gene interaction network and environmental chemical-gene interaction network were established, and Cytoscape 3.72 was used to perform visualization. RESULTS: After data standardization, the data quality was excellent and it can ensure reliable results. Myocardial cell infection with SARS-CoV-2 and gene expression spectrum were changed significantly, including a total of 484 DEGs in adult cardiomyoblasts, a total of 667 DEGs in macrophages, and a total of 1 483 DEGs in human embryo source of cardiomyopathy. The Stum, mechanosensory transduction mediator homolog (STUM), dehydrogenase/reductase 9 (DHRS9), calcium/calmodulin dependent protein kinase II beta (CAMK2B), claudin 1(CLDN1), C-C motif chemokine ligand 2 (CCL2), TNFAIP3 interacting protein 3 (TNIP3), G protein-coupled receptor 84 (GPR84), and C-X-C motif chemokine ligand 1 (CXCL1) were identical in expression patterns in 3 types of cells. The protein-protein interaction suggested that CAMK2B proteins may play a key role in the antiviral process in 3 types of cells; and silicon dioxide (SiO2), benzodiazepine (BaP), nickel (Ni), and estradiol (E2) affect anti-SARS-CoV-2 processes of the 3 types of cells. CONCLUSIONS: CAMK2B, CLDN1, CCL2, and DHRS9 genes play important roles in the immune response of cardiomyocytes against SARS-CoV-2. SiO2, BaP, Ni, E2 may affect the cell's antiviral process by increasing the toxicity of cardiomyocytes, thereby aggravating SARS-CoV-2 harm to the heart.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , Macrophages , Myocytes, Cardiac , Silicon Dioxide , Transcriptome
8.
Int J Nanomedicine ; 16: 6575-6591, 2021.
Article in English | MEDLINE | ID: covidwho-1533527

ABSTRACT

Public awareness of infectious diseases has increased in recent months, not only due to the current COVID-19 outbreak but also because of antimicrobial resistance (AMR) being declared a top-10 global health threat by the World Health Organization (WHO) in 2019. These global issues have spiked the realization that new and more efficient methods and approaches are urgently required to efficiently combat and overcome the failures in the diagnosis and therapy of infectious disease. This holds true not only for current diseases, but we should also have enough readiness to fight the unforeseen diseases so as to avoid future pandemics. A paradigm shift is needed, not only in infection treatment, but also diagnostic practices, to overcome the potential failures associated with early diagnosis stages, leading to unnecessary and inefficient treatments, while simultaneously promoting AMR. With the development of nanotechnology, nanomaterials fabricated as multifunctional nano-platforms for antibacterial therapeutics, diagnostics, or both (known as "theranostics") have attracted increasing attention. In the research field of nanomedicine, mesoporous silica nanoparticles (MSN) with a tailored structure, large surface area, high loading capacity, abundant chemical versatility, and acceptable biocompatibility, have shown great potential to integrate the desired functions for diagnosis of bacterial infections. The focus of this review is to present the advances in mesoporous materials in the form of nanoparticles (NPs) or composites that can easily and flexibly accommodate dual or multifunctional capabilities of separation, identification and tracking performed during the diagnosis of infectious diseases together with the inspiring NP designs in diagnosis of bacterial infections.


Subject(s)
Bacterial Infections , COVID-19 , Nanoparticles , Bacterial Infections/diagnosis , Bacterial Infections/drug therapy , Humans , Porosity , SARS-CoV-2 , Silicon Dioxide
9.
Environ Sci Pollut Res Int ; 29(12): 17001-17016, 2022 Mar.
Article in English | MEDLINE | ID: covidwho-1469752

ABSTRACT

The balanced amelioration of mechanical characteristics of fat clay with an additive refers to the attainment of high strength without compromising ductility, which is unattainable by solitary usage of a cementing additive. For this purpose, an amalgamated binary admixture (ABA) is proposed by assimilating shredded face mask (FM) waste, which is posing serious environmental concerns these days, with a cementitious waste material, i.e., silica fume (SF). However, for such ABA, the optimization of mix design is desirable because an excessive amount of one component could disturb the required balance. To address this issue, response surface methodology (RSM) is used in the current study, which is a strong technique used during the process of production to develop, improve, and optimize product inputs. Several experiments are designed and conducted to evaluate mechanical responses, i.e., unconfined compressive strength (qu), brittleness index (IB), deformability index (ID), and California bearing ratio (CBR) value, of treated fat clay by varying mix designs of ABA. Based on the test results, mathematical models are developed which are found to be statistically valid to predict the subjected responses using SF and FM as inputs. Afterward, an optimized mix design is determined by integrating developed models with a desirability function model and setting maximization of strength and ductility as the optimization goals. An ABA having 7.9% SF and 1.2% FM is observed to provide the highest strength and ductility for multiple applications, i.e., road and buildings, with desirability factor close to unity; responses of which are also validated by performing tests. Furthermore, analysis of cleaning aspect shows that the use of optimized ABA in place of cement for subgrade improvement of 1 km two-lane road could avoid CO2 emission of around 79,032 kg of C, save 42,720 kWh and 1174.8 GJ of electrical and thermal energy, respectively, and clean 43 Mg of FM waste; however, astute protocols of COVID-19 FM waste handling and disinfection are needed to be established and followed.


Subject(s)
COVID-19 , Masks , Clay , Humans , SARS-CoV-2 , Silicon Dioxide
10.
Int J Mol Sci ; 22(19)2021 Oct 08.
Article in English | MEDLINE | ID: covidwho-1463712

ABSTRACT

Nanomaterials have found extensive interest in the development of novel vaccines, as adjuvants and/or carriers in vaccination platforms. Conjugation of protein antigens at the particle surface by non-covalent adsorption is the most widely used approach in licensed particulate vaccines. Hence, it is essential to understand proteins' structural integrity at the material interface in order to develop safe-by-design nanovaccines. In this study, we utilized two model proteins, the wild-type allergen Bet v 1 and its hypoallergenic fold variant (BM4), to compare SiO2 nanoparticles with Alhydrogel® as particulate systems. A set of biophysical and functional assays including circular dichroism spectroscopy and proteolytic degradation was used to examine the antigens' structural integrity at the material interface. Conjugation of both biomolecules to the particulate systems decreased their proteolytic stability. However, we observed qualitative and quantitative differences in antigen processing concomitant with differences in their fold stability. These changes further led to an alteration in IgE epitope recognition. Here, we propose a toolbox of biophysical and functional in vitro assays for the suitability assessment of nanomaterials in the early stages of vaccine development. These tools will aid in safe-by-design innovations and allow fine-tuning the properties of nanoparticle candidates to shape a specific immune response.


Subject(s)
Allergens/immunology , Antigens, Plant/immunology , Epitopes/immunology , Lymphocyte Activation/immunology , Nanoparticles/chemistry , Silicon Dioxide/chemistry , Vaccines/immunology , Allergens/chemistry , Humans , Hydrogels , Immunoglobulin E/immunology , Respiratory Hypersensitivity/immunology , T-Lymphocytes/immunology
11.
Biomater Sci ; 9(21): 7287-7296, 2021 Oct 26.
Article in English | MEDLINE | ID: covidwho-1454827

ABSTRACT

Development of a rapidly scalable vaccine is still an urgent task to halt the spread of COVID-19. We have demonstrated biodegradable mesoporous silica nanoparticles (BMSNs) as a good drug delivery carrier for tumor therapy. In this study, seven linear B cell epitopes and three CD8+ T cell epitopes were screened from the spike (S) glycoprotein of SARS-CoV-2 by computer-based immunoinformatic approaches for vaccine design. A nanoparticle-based candidate vaccine (B/T@BMSNs) against SARS-CoV-2 was rapidly prepared by encapsulating these ten epitope peptides within BMSNs, respectively. BMSNs with potential biodegradability, proved to possess excellent safety in vitro and in vivo, could efficiently deliver epitope peptides into the cytoplasm of RAW264.7 cells. Strong Th1-biased humoral and cellular immunity were induced by B/T@BMSNs in mice and all the 10 selected epitopes were identified as effective antigen epitopes, which could induce robust peptide-specific immune response. The elicited functional antibody could bind to the recombinant S protein and block the binding of the S protein to the ACE-2 receptor. These results demonstrate the potential of a nanoparticles vaccine platform based on BMSNs to rapidly develop peptide-based subunit vaccine candidates against SARS-CoV-2.


Subject(s)
COVID-19 , Nanoparticles , Animals , COVID-19 Vaccines , Epitopes, T-Lymphocyte , Humans , Immunity, Cellular , Mice , Peptides , SARS-CoV-2 , Silicon Dioxide , Spike Glycoprotein, Coronavirus , Vaccines, Subunit
12.
Mater Sci Eng C Mater Biol Appl ; 130: 112438, 2021 Nov.
Article in English | MEDLINE | ID: covidwho-1415666

ABSTRACT

Recently, great attention has been paid to hydroxychloroquine which after promising in vitro studies has been proposed to treat the severe acute respiratory syndrome caused by SARS-CoV-2. The clinical trials have shown that hydroxychloroquine was not as effective as was expected and additionally, several side effects were observed in patients cured with this medicament. In order to reduce them, it is suggested to deliver hydroxychloroquine in a controlled manner. Therefore, in this study non-modified (SBA-15, SBA-16) and modified with copper and aminosilane mesoporous silica materials were applied as novel nanocarriers for hydroxychloroquine. First, pristine and functionalized samples were synthesized and characterized by X-ray diffraction, low-temperature nitrogen sorption, transmission electron microscopy, X-ray photoelectron spectroscopy, infrared spectroscopy, laser diffraction. Then the influence of physicochemical parameters of materials obtained on the adsorption and release processes of hydroxychloroquine was analyzed. The mechanism of hydroxychloroquine binding to non-modified silicas was based on the formation of hydrogen bonds, while in the case of copper and aminosilane functionalized materials the complexes with drug molecules were generated. The release behavior of hydroxychloroquine from silica samples obtained was determined by different factors including pH conditions, textural parameters, surface charge, and presence of surface functional groups. The greatest differences in hydroxychloroquine release profiles between materials were observed at pH 7.2. The amount of drug desorbed from silica decreased in the following order: functionalized SBA-15 (84%) > functionalized SBA-16 (79%) > SBA-15 (59%) > SBA-16 (33%). It proved that a higher amount of drug was released from materials of hexagonal structure.


Subject(s)
COVID-19 , Copper , COVID-19/drug therapy , Humans , Hydroxychloroquine , SARS-CoV-2 , Silicon Dioxide
13.
Biotechniques ; 71(4): 510-515, 2021 10.
Article in English | MEDLINE | ID: covidwho-1411518

ABSTRACT

Purity and integrity are two important criteria for any RNA extraction process to qualify the RNA for meaningful gene expression analysis. This study compares four commercially available RNA extraction kits using silica membrane and magnetic bead separation methods. The performance was evaluated in terms of both quantity (total RNA amount in µg/µl) and purity (260/280 ratio). The concentration and purity of each kit was significantly different from those of the others (p < 0.001). Although quantity obtained from Mag MAX is comparatively lower than QIAGEN, the quality is comparable as evident from real-time PCR performance. This study suggests that there are practical differences between these RNA extraction kits that should be taken into account while isolating RNA required for gene expression analysis.


Subject(s)
Magnets/chemistry , Membranes, Artificial , RNA, Viral/isolation & purification , Reagent Kits, Diagnostic , Silicon Dioxide/chemistry , COVID-19/diagnosis , COVID-19/virology , COVID-19 Nucleic Acid Testing/methods , Gene Expression Profiling/methods , Humans , Polymerase Chain Reaction/methods , RNA, Viral/genetics , SARS-CoV-2/genetics , SARS-CoV-2/isolation & purification
14.
Mol Biol Rep ; 48(10): 6871-6877, 2021 Oct.
Article in English | MEDLINE | ID: covidwho-1401056

ABSTRACT

BACKGROUND: RT-qPCR technique is the current world-wide method used for the early detection of SARS-CoV2 RNA in the suspected clinical samples. Viral RNA extraction is the key pre-analytical step for SARS-CoV2 detection which often achieved using commercial RNA-extraction kits. However, due to the COVID-19 pandemic, bulk production and the supply chains for the commercial RNA-extraction kit have been seriously compromised. The shortage of commercial RNA-extraction kit is even more acute in developing country. Furthermore, use of one-off design RNA-columns can generate plastic wastes that have an environmental pollution effect. METHODS AND RESULTS: To address these issues, in this study, we used warm alkaline solution containing Triton X-100 for the complete removal of the residual SARS-CoV2 RNA from the used RNA-binding silica column. Columns regenerated using the alkaline solution have the viral RNA purification capability that is comparable to the fresh silica columns. We also demonstrated that RNA-binding silica columns can be regenerated and reused for a minimum of five-times. CONCLUSIONS: Therefore, the use of the RNA-column regeneration method may benefits several SARS-CoV2 diagnostic laboratories throughout the world by cutting down the requirement of commercial RNA-purification column.


Subject(s)
COVID-19 Nucleic Acid Testing/instrumentation , Chromatography/instrumentation , RNA, Viral/isolation & purification , COVID-19 Nucleic Acid Testing/methods , Chromatography/methods , Humans , Octoxynol , RNA, Viral/metabolism , Real-Time Polymerase Chain Reaction , Recycling , Silicon Dioxide
15.
ACS Sens ; 6(9): 3468-3476, 2021 09 24.
Article in English | MEDLINE | ID: covidwho-1392782

ABSTRACT

This research reveals the promising functionalization of graphene oxide (GrO)-glazed double-interdigitated capacitive (DIDC) biosensing platform to detect severe acute respiratory syndrome coronavirus (SARS-CoV-2) spike (S1) proteins with enhanced selectivity and rapid response. The DIDC bioactive surface consisting of Pt/Ti featured SiO2 substrate was fabricated using GrO/EDC-NHS/anti-SARS-CoV-2 antibodies (Abs) which is having layer-by-layer interface self-assembly chemistry method. This electroactive immune-sensing platform exhibits reproducibility and sensitivity with reference to the S1 protein of SARS-CoV-2. The outcomes of analytical studies confirm that GrO provided a desired engineered surface for Abs immobilization and amplified capacitance to achieve a wide detection range (1.0 mg/mL to 1.0 fg/mL), low limit of detection (1 fg/mL) within 3 s of response time, good linearity (18.56 nF/g), and a high sensitivity of 1.0 fg/mL. Importantly, the unique biochip was selective against blood-borne antigens and standby for 10 days at 5 °C. Our developed DIDC-based SARS-CoV-2 biosensor is suitable for point-of-care (POC) diagnostic applications due to portability and scaling-up ability. In addition, this sensing platform can be modified for the early diagnosis of severe viral infections using real samples.


Subject(s)
COVID-19 , SARS-CoV-2 , Graphite , Humans , Reproducibility of Results , Silicon Dioxide , Spike Glycoprotein, Coronavirus
16.
Biophys Chem ; 265: 106441, 2020 10.
Article in English | MEDLINE | ID: covidwho-1343142

ABSTRACT

The possibility of immobilizing a protein with antigenic properties on a solid support offers significant possibilities in the development of immunosensors and vaccine formulations. For both applications, the orientation of the antigen should ensure ready accessibility of the antibodies to the epitope. However, an experimental assessment of the orientational preferences necessarily proceeds through the preparation/isolation of the antigen, the immobilization on different surfaces and one or more biophysical characterization steps. To predict a priori whether favorable orientations can be achieved or not would allow one to select the most promising experimental routes, partly mitigating the time cost towards the final product. In this manuscript, we apply a simple computational model, based on united-residue modelling, to the prediction of the orientation of the receptor binding domain of the SARS-CoV-2 spike protein on surfaces commonly used in lateral-flow devices. These calculations can account for the experimental observation that direct immobilization on gold gives sufficient exposure of the epitope to obtain a response in immunochemical assays.


Subject(s)
Betacoronavirus/metabolism , Epitopes/chemistry , Models, Molecular , Spike Glycoprotein, Coronavirus/metabolism , Antigens/chemistry , Antigens/immunology , Antigens/metabolism , Epitopes/immunology , Molecular Docking Simulation , Protein Domains , SARS-CoV-2 , Silicon Dioxide/chemistry , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/immunology , Surface Properties
17.
J Control Release ; 337: 14-26, 2021 09 10.
Article in English | MEDLINE | ID: covidwho-1306406

ABSTRACT

Acute lung injury (ALI) is a critical inflammatory syndrome, characterized by increased diffuse inflammation and severe lung damage, which represents a clinical concern due to the high morbidity and mortality in critical patients. In last years, there has been a need to develop more effective treatments for ALI, and targeted drug delivery to inflamed lungs has become an attractive research field. Here, we present a nanodevice based on mesoporous silica nanoparticles loaded with dexamethasone (a glucocorticoid extensively used for ALI treatment) and capped with a peptide that targets the TNFR1 receptor expressed in pro-inflammatory macrophages (TNFR-Dex-MSNs) and avoids cargo leakage. TNFR-Dex-MSNs nanoparticles are preferentially internalized by pro-inflammatory macrophages, which overexpressed the TNFR1 receptor, with the subsequent cargo release upon the enzymatic hydrolysis of the capping peptide in lysosomes. Moreover, TNFR-Dex-MSNs are able to reduce the levels of TNF-α and IL-1ß cytokines in activated pro-inflammatory M1 macrophages. The anti-inflammatory effect of TNFR-Dex-MSNs is also tested in an in vivo ALI mice model. The administered nanodevice (intravenously by tail vein injection) accumulated in the injured lungs and the controlled dexamethasone release reduces markedly the inflammatory response (TNF-α IL-6 and IL-1ß levels). The attenuation in lung damage, after treatment with TNFR-Dex-MSNs, is also confirmed by histopathological studies. Besides, the targeted-lung dexamethasone delivery results in a decrease of dexamethasone derived side-effects, suggesting that targeted nanoparticles can be used for therapy in ALI and could help to overcome the clinical limitations of current treatments.


Subject(s)
Acute Lung Injury , Nanoparticles , Acute Lung Injury/drug therapy , Animals , Dexamethasone , Humans , Lung , Mice , Silicon Dioxide
18.
Analyst ; 146(12): 3908-3917, 2021 Jun 14.
Article in English | MEDLINE | ID: covidwho-1319050

ABSTRACT

The pandemic outbreak of the 2019 coronavirus disease (COVID-19), which is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is still spreading rapidly and poses a great threat to human health. As such, developing rapid and accurate immunodiagnostic methods for the identification of infected persons is needed. Here, we proposed a simple but sensitive on-site testing method based on spike protein-conjugated quantum dot (QD) nanotag-integrated lateral flow immunoassay (LFA) to simultaneously detect SARS-CoV-2-specific IgM and IgG in human serum. Advanced silica-core@dual QD-shell nanocomposites (SiO2@DQD) with superior luminescence and stability were prepared to serve as fluorescent nanotags in the LFA strip and guarantee high sensitivity and reliability of the assay. The performance of the SiO2@DQD-strip was fully optimized and confirmed by using 10 positive serum samples from COVID-19 patients and 10 negative samples from patients with other respiratory diseases. The practical clinical value of the assay was further evaluated by testing 316 serum samples (114 positive and 202 negative samples). The overall detection sensitivity and specificity reached 97.37% (111/114) and 95.54% (193/202), respectively, indicating the huge potential of our proposed method for the rapid and accurate detection of SARS-CoV-2-infected persons and asymptomatic carriers.


Subject(s)
COVID-19 , Spike Glycoprotein, Coronavirus , Antibodies, Viral , Humans , Immunoassay , Immunoglobulin G , Immunoglobulin M , Reproducibility of Results , SARS-CoV-2 , Sensitivity and Specificity , Silicon Dioxide
19.
Anal Chem ; 93(30): 10444-10452, 2021 08 03.
Article in English | MEDLINE | ID: covidwho-1319008

ABSTRACT

The COVID-19 pandemic caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious public health threat. Most vaccines against SARS-CoV-2 target the highly glycosylated spike protein (S). A good knowledge of the glycosylation profile of this protein is key to successful vaccine development. Unlike the 22 confirmed N-glycosylation sites on SARS-CoV-2 S, only a few O-glycosylation sites on this protein have been reported. This difference is mainly ascribed to the extremely low stoichiometry of O-glycosylation. Herein, we designed the biomimetic materials, Trp-Arg (WR) monomer-grafted silica microspheres (designated as WR-SiO2), and these biomimetic materials can enrich N- and O-linked glycopeptides with high selectivity. And WR-SiO2 can resist the nonglycopeptides' interference with the 100 molar fold of BSA during O-linked glycopeptide enrichment. We utilized WR-SiO2 to comprehensively analyze the O-glycosylation profile of recombinant SARS-CoV-2 S. Twenty-seven O-glycosylation sites including 18 unambiguous sites are identified on SARS-CoV-2 S. Our study demonstrates that the biomimetic polymer can offer specific selectivity for O-linked glycopeptides and pave the way for O-glycosylation research in biological fields. The O-glycosylation profile of SARS-CoV-2 S might supplement the comprehensive glycosylation in addition to N-glycosylation of SARS-CoV-2 S.


Subject(s)
Biomimetic Materials , COVID-19 , Biomimetics , COVID-19 Vaccines , Glycosylation , Humans , Pandemics , SARS-CoV-2 , Silicon Dioxide , Spike Glycoprotein, Coronavirus/metabolism
20.
Analyst ; 146(16): 5055-5060, 2021 Aug 09.
Article in English | MEDLINE | ID: covidwho-1317820

ABSTRACT

It is imminent to develop a new type of rapid COVID-19 detection method with high sensitivity. Here, we used novel red emission-enhanced carbon dot (CD)-based silica (RCS) spheres as the signals of lateral flow immunochromatography (LFI) to ultrasensitively detect novel severe acute respiratory syndrome coronavirus 2 nucleocapsid proteins (SARS-CoV-2 NPs). The red emission of CDs can be enhanced and enriched in silica spheres by a simple way. The amino ends of the N-ß-(aminoethyl)-γ-aminopropyltrimethoxy anchor carboxyl-rich CDs and enhance the red emission, while the other end is embedded in the silica carrier. Then, the composite silica spheres werecoated with 3-(triethylsilyl) propylamine to protect the CDs, promote bioconjugation and obtain RCS spheres. The optimal emission peaks of the aqueous solution and the solid state of RCS spheres were at 634 nm and 638 nm, respectively, with quantum yields (QYs) of 48.5% and 35.7%, respectively. Their red emission has a wide excitation range (from the ultraviolet region to the red region), and the best excitation wavelength is about 580 nm. Two fluorescence detection modes of the RCS-LFI technology for the SARS-CoV-2 NP assay are available: the simple mode of observation under ultraviolet light has a sensitivity of 100 pg mL-1; the advanced mode of detection under a fluorescence microscope has a sensitivity of 10 pg mL-1. This assay also exhibits the advantages of fast detection speed, high specificity, and simple operation. In addition, the feasibility of this method in actual sample detection was verified in human serum by the standard-addition method, and the results show that the method has excellent practicability. We believe that this method will be a valuable supplement for the diagnosis of COVID-19.


Subject(s)
COVID-19 , SARS-CoV-2 , Antibodies, Viral , Carbon , Humans , Immunoassay , Nucleocapsid Proteins , Sensitivity and Specificity , Silicon Dioxide
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