Biosensors-Basel
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2024
·Vol. 14
·期 7
·332
·
IF 4.9
Xiaonan Zhang,Lizhong Zhang,Han Li,Yang Xu,Lingqin Meng,Gengyu Liang,Bei Wang,Le Liu,Tian Guan,Cuixia Guo,Yonghong He
The demand for accurate and efficient immunoassays calls for the development of precise, high-throughput analysis methods. This paper introduces a novel approach utilizing a weak measurement interface sensor for immunoassays, offering a solution for high throughput analysis. Weak measurement is a precise quantum measurement method that amplifies the weak value of a system in the weak interaction through appropriate pre- and post-selection states. To facilitate the simultaneous analysis of multiple samples, we have developed a chip with six flow channels capable of conducting six immunoassays concurrently. We can perform real-time immunoassay to determine the binding characteristics of spike protein and antibody through real-time analysis of the flow channel images and calculating the relative intensity. The proposed method boasts a simple structure, eliminating the need for intricate nano processes. The spike protein concentration and relative intensity curve were fitted using the Log-Log fitting regression equation, and R2was 0.91. Utilizing a pre-transformation approach to account for slight variations in detection sensitivity across different flow channels, the present method achieves an impressive limit of detection(LOD) of 0.85 ng/mL for the SARS-CoV-2 the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein, with a system standard deviation of 5.61. Furthermore, this method has been successfully verified for monitoring molecular-specific binding processes and differentiating binding capacities.
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OPTICAL MATERIALS
·
2025
·
IF 4.2
Bing Xu,Xiaoying Xiang,Yan Li,Liu Yang,Zhihui Luo,Jianglou Huang
To tackle the demand for highly accurate and real-time diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a highly sensitive optical fiber SARS-CoV-2 antigen sensor based on triangular gold nanosheets/gold film coupling is proposed. The coupling effect between the local surface plasmon resonance effect of the triangular gold nanosheets and the surface plasmon resonance effect of the gold film increases the local electric field, resulting in higher sensitivity and lower detection limit of the sensor, and the refractive index sensitivity of the sensor is increased to 3894.3 nm/RIU. The antigen reacts specifically with the viral antibody adsorbed on the surface of the optical fiber, which leads to a small change in refractive index, which ultimately leads to a shift in the resonance peak of the optical fiber, thus enabling antigen detection. The biosensor showed good response in the range of SARS-CoV-2 antigen concentration from 0.035 to 100 ng/mL, and the limit of detection (LOD) was 0.017 ng/mL (S/N=3). The biosensor has good selectivity and stability, possesses a short response time (10 min), which provides a new idea for the detection of SARS-CoV-2 in the biomedical field.
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BIOSENSORS & BIOELECTRONICS
·
2025
·
IF 10.5
Hang Ao,Jinshan Xiong,Wencheng Xiao,Wenrui Hu,Jie Wu,Huangxian Ju
Owing to the continuous mutations of the coronavirus, the sensitive and accurate detection of the SARS-CoV-2 nucleocapsid protein is still necessary in clinical diagnosis. Aim to the challenge of sensitivity in chemiluminescence (CL) imaging detection and the need for high-throughput clinical screening, this work used imidazole (Im) as an enhancer for hemin-DNA/luminol/H 2 O 2 CL system to propose a CL imaging method for SARS-CoV-2 nucleocapsid protein detection. In this system Im could reduce the hemin-DNA catalyzed oxidation energy of luminol by H 2 O 2 to promote the CL emission through the accelerated breaking of O-O bond of dianionic cyclic peroxide. The enhancement mechanism was demonstrated with theoretical calculations and several characterization techniques. By combining the proximity-induced initiator release with immunological recognition, an initiator-triggered DNA nanomachine was designed to achieve the release of abundant primers for activating hemin-DNA switch. This signal amplification strategy led to strong CL emission for highly sensitive imaging detection of the target. The Im-enhanced hemin-DNA/luminol/H 2 O 2 CL imaging assay exhibited a linear detection range over 4 orders of magnitude with a detection limit down to 2.75 pg/mL for SARS-CoV-2 nucleocapsid protein, exhibiting promising potential of both Im sensitization and the proposed DNA nanomachine to improve the sensitivity of high-throughput CL biosensing for the early diagnosis and clinical screening of the coronavirus and other diseases.
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Molecular Medicine Reports
·
2024
·Vol. 29
·期 3
·1-12
·
IF 3.4
Mi-Hua Liu,Xiao-Long Lin,Le-Le Xiao
Hyperphosphatemia or severe acute respiratory syndrome coronavirus 2 (SARS‑CoV‑2) infection can promote cardiovascular adverse events in patients with chronic kidney disease. Hyperphosphatemia is associated with elevated inflammation and sterol regulatory element binding protein 2 (SREBP2) activation, but the underlying mechanisms in SARS‑CoV‑2 that are related to cardiovascular disease remain unclear. The present study aimed to elucidate the role of excess inorganic phosphate (PI) in SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation and the underlying mechanisms in vascular smooth muscle cells (VSMCs). The expression levels of SARS‑CoV‑2 N protein, SREBP cleavage‑activating protein (SCAP), mature N‑terminal SREBP2, NLRP3, procaspase‑1, cleaved caspase‑1, IL‑1β and IL‑18 were examined by western blotting. The expression levels of SREBP2, HMG‑CoA reductase, HMGCS1, low density lipoprotein receptor, proprotein convertase subtilisin/kexin type 9 (PCSK9), SREBP1c, fatty acid synthase, stearyl coenzyme A desaturase 1, acetyl‑CoA carboxylase α and ATP‑citrate lyase were determined by reverse transcription‑quantitative PCR. The translocation of SCAP or NLRP3 from the endoplasmic reticulum to the Golgi was detected by confocal microscopy. The results showed that excess PI promoted SCAP‑SREBP and NLRP3 complex translocation to the Golgi, potentially leading to NLRP3 inflammasome activation and lipogenic gene expression. Furthermore, PI amplified SARS‑CoV‑2 N protein‑induced inflammation via the SCAP‑SREBP pathway, which facilitates NLRP3 inflammasome assembly and activation. Inhibition of phosphate uptake with phosphonoformate sodium alleviated NLRP3 inflammasome activation and reduced SREBP‑mediated lipogenic gene expression in VSMCs stimulated with PI and with SARS‑CoV‑2 N protein overexpression. Inhibition of SREBP2 or small interfering RNA‑induced silencing of SREBP2 effectively suppressed the effect of PI and SARS‑CoV‑2 N protein on NLRP3 inflammasome activation and lipogenic gene expression. In conclusion, the present study identified that PI amplified SARS‑CoV‑2 N protein‑induced NLRP3 inflammasome activation and lipogenic gene expression via the SCAP‑SREBP signaling pathway.
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ANALYST
·
2025
·
IF 3.3
Yang Li,Yikan Zhao,Zhihao Yi,Shitong Han
Timely and accurate diagnosis of RNA viruses, represented by the SARS-CoV-2, is the foundation for protecting people from health threats. Currently, direct detection of viral RNA genes remains the most accurate method. Herein, a rapid, ultrasensitive, and no heating equipment required CRISPR/Cas13a based evanescent wave fluorescence biosensing platform for quantitative detection of the SARS-CoV-2 gene is reported. The collateral effect of CRISPR/Cas13a for RNA is combined with a self-driven enzyme-free hybridization chain reaction (HCR) as a signal amplification step. When the initiator RNA strand is cleaved by Cas13a, the downstream signal amplification induced by HCR is blocked, and a multiple crRNA strategy is used to enhance the cleavage efficiency. The newly designed HCR assemblies are captured by the cDNA-modified optical fiber and generate a higher-intensity fluorescence signal induced by the evanescent field. The CRISPR/Cas13a-HCR evanescent wave fluorescence biosensing platform is capable of detection of SARS-CoV-2 with a LOD of 0.47 copies per μL and the detection time is within 35 min. The spike recovery tests in saliva and high specificity have demonstrated the potential of this method for point-of-care diagnosis. This method is also suitable for the detection of other RNA viruses, without the need to alter any design of the HCR component, and only the corresponding crRNA needs to be replaced.
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ChemistrySelect
·
2025
·Vol. 10
·期 15
·e202406023
·
IF 2
Leyao Chen,Nuoya Wang,Xiaoyu Chang,Ruiyong Wang
Graphical This work investigated the inhibition of 4-thiazolidinone derivatives on SARS-CoV-2 Mpro using enzyme assays, spectroscopy, and molecular docking. Compound 3a showed the best inhibition (IC 50 = 2.36 ± 0.36 µM, K i = 1.72 µM), and the trifluoromethyl group in 3a played an important role. Hydrogen bonding and van der Waals forces were the main forces for the binding. This work provided clues for Mpro inhibitor development. Since the outbreak of COVID-19, one of the strategies used to search for potential drugs has been to screen inhibitors of SARS-CoV-2 Mpro. This work investigated the inhibitory effects of 4-thiazolidinone derivatives on main protease (Mpro) using various methods, including enzyme activity assays, enzyme kinetics, spectroscopy experiments, molecular docking simulations, microscale thermophoresis, and cytotoxicity tests. Compound 3a demonstrated the best inhibition effect among the compounds tested, with IC 50 = 2.36 ± 0.36 µM and K i = 1.72 µM. The trifluoromethyl group in 3a might enhance its inhibition ability. Interactions between the 4-thiazolidinone derivatives and Mpro resulted in fluorescence quenching primarily through static quenching. Thermodynamic analysis indicated that the interaction forces are mainly hydrogen bonds and van der Waals forces. Spectral results revealed that the 4-thiazolidinone derivatives affected the microenvironment of aromatic amino acid residues in Mpro and altered its secondary structure. Differential scanning fluorimetry was used to confirm the interaction between the 4-thiazolidinone derivatives and Mpro. Results of microscale thermophoresis indicated that the K d between 3a and Mpro was 1.56 ± 0.24 µM. Molecular docking simulations provided a more intuitive visualization of the interaction forces. This study provides clues for screening novel Mpro inhibitors.
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TALANTA
·
2025
·
IF 6.1
Lingyan Kang,Tao Xu,Zhimeng Fan,Yiqi Li,Ye Tian
Achieving multiplexed, equipment-free, and sensitivity-enhanced antigen detection within a true one-step lateral flow format remains a technical challenge. Lateral flow immunoassays (LFIAs) are widely used for point-of-care diagnostics, yet their sensitivity—particularly in weak-positive specimens—often limits reliable pathogen identification. Herein, we enhance the analytical performance of LFIA by introducing a structurally integrated self-enhancement strategy that maintains the simplicity of one-step operation without requiring additional user steps. In the proposed SET-LFIA, two enhancement pads loaded with Au 3+ and NH 2 OH·HCl are positioned beneath the detection layer and isolated by a double-sided adhesive barrier, enabling sequential reagent release along the capillary flow path and inducing localized in situ gold growth during the post-binding phase. This design strengthens the visual signal while preserving the immunoreaction process. Under optimized conditions, the SET-LFIA achieved detection limits of 1.0 ng/mL for IAV, 1.0 ng/mL for IBV, and 0.2 ng/mL for SARS-CoV-2, corresponding to 5–25-fold sensitivity gains over conventional LFIA. Tests using spiked samples demonstrated markedly improved discrimination of weak-positive specimens and reduced false-negative outcomes. In addition, stability evaluation showed >85 % signal retention after 21 days at 45 °C, and clinical samples further confirmed robust multiplex detection across all three targets. With its enhanced sensitivity, one-step workflow, and compatibility with industrial lamination, the SET-LFIA represents a practical and scalable solution for rapid on-site respiratory pathogen screening.
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Polymers
·
2022
·Vol. 14
·期 24
·5466
·
IF 4.967
Changcai Teng,Xiongyan Meng,Yeqin Hu,Hongzhao Mao,Huiting Li,Jing Yang,Tiantian Sun,Shuai Meng,Chengli Zong
Small synthetic TLR7/8-agonists can be used as vaccine adjuvants to enhance cell and humoral-mediated immune responses to specific antigens. Despite their potency, after local injection they can be dispersed to undesired body parts causing high reactogenicity, limiting their clinical applications. Here we describe a vaccination strategy that employs the covalent conjugate of a mannose and TLR7/8 agonist as a vaccine adjuvant to take advantage of mannose binding C-type lectins on dendritic cells to enhance the vaccine’s immunogenicity. The mannose-TLR7/8 agonist conjugate can self-assemble into nanoparticles with the hydrophilic mannose on the outside and hydrophobic TLR7/8 agonist inside. Although its ability to stimulate HEK-BlueTMhTLR7/8 cells dropped, it can efficiently stimulate mouse bone marrow-derived dendritic cells as indicated by the up-regulation of CD80 and CD86, and higher cytokine expression levels of TNF-α, IL6, and IL-12p70 than the native TLR7/8 agonist. In vivo, vaccination using the SARS-CoV-2 RBD trimer as the antigen and the conjugate as the adjuvant induced a significantly higher amount of IgG2a. These results suggest that the mannose-TLR7/8-agonist conjugate can be used as an effective vaccine adjuvant.Keywords:adjuvants;mannose;TLR7/8 agonist;immune responses;cytokines;antibody
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MICROCHIMICA ACTA
·
2021
·Vol. 188
·期 10
·1-10
·
IF 5.833
Chen Rongyuan,Kan Lun,Duan Fenghe,He Linghao,Wang Minghua,Cui Jing,Zhang Zhihong,Zhang Zhonghou
A novel label-free surface plasmon resonance (SPR) aptasensor has been constructed for the detection of N-gene of SARS-CoV-2 by using thiol-modified niobium carbide MXene quantum dots (Nb 2 C-SH QDs) as the bioplatform for anchoring N-gene-targeted aptamer. In the presence of SARS-CoV-2 N-gene, the immobilized aptamer strands changed their conformation to specifically bind with N-gene. It thus increased the contact area or enlarged the distance between aptamer and the SPR chip, resulting in a change of the SPR signal irradiated by the laser (He-Ne) with the wavelength ( λ ) of 633 nm. Nb 2 C QDs were derived from Nb 2 C MXene nanosheets via a solvothermal method, followed by functionalization with octadecanethiol through a self-assembling method. Subsequently, the gold chip for SPR measurements was modified with Nb 2 C-SH QDs via covalent binding of the Au-S bond also by self-assembling interaction. Nb 2 C-SH QDs not only resulted in high bioaffinity toward aptamer but also enhanced the SPR response. Thus, the Nb 2 C-SH QD-based SPR aptasensor had low limit of detection (LOD) of 4.9 pg mL −1 toward N-gene within the concentration range 0.05 to 100 ng mL −1 . The sensor also showed excellent selectivity in the presence of various respiratory viruses and proteins in human serum and high stability. Moreover, the Nb 2 C-SH QD-based SPR aptasensor displayed a vast practical application for the qualitative analysis of N-gene from different samples, including seawater, seafood, and human serum. Thus, this work can provide a deep insight into the construction of the aptasensor for detecting SARS-CoV-2 in complex environments. Graphical abstract
A novel label-free surface plasmon resonance aptasensor has been constructed to detect sensitively and selectively the N-gene of SARS-CoV-2 by using thiol-modified niobium carbide MXene quantum dots as the scaffold to anchor the N-gene-targeted aptamer.
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BIOORGANIC CHEMISTRY
·
2021
·
IF 5.275
Xinjian Yin,Litong Chen,Siwen Yuan,Lan Liu,Zhizeng Gao
Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is a serious threat to global health. One attractive antiviral target is the membrane fusion mechanism employed by the virus to gain access to the host cell. Here we report a robust protein-based fluorescent polarization assay, that mimicking the formation of the six-helix bundle (6-HB) process during the membrane fusion, for the evaluation and screening of SARS-CoV-2 fusion Inhibitors. The IC 50 of known inhibitors, HR2P, EK1, and Salvianolic acid C (Sal-C) were measured to be 6.1 nM, 2.5 nM, and 8.9 µM respectively. In addition, we found Sal-A has a slightly lower IC 50 (3.9 µM) than Sal-C. Interestingly, simple caffeic acid can also disrupt the formation of 6-HB with a sub-mM concentration. Pilot high throughput screening (HTS) of a small marine natural product library validates the assay with a Z’ factor close to 0.8. We envision the current assay provides a convenient way to screen SARS-CoV-2 fusion inhibitors and assess their binding affinity.
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STAR Protocols
·
2022
·
IF 0
Jing Zhang,Haohao Yan,Gangan Yan,Xiaoping Liu,Yanchang Wang,Yunyu Chen
Summary Discovery of efficacious antiviral agents targeting SARS-CoV-2 main protease (Mpro) is of the highest importance to fight against COVID-19. Here, we describe a simple protocol for high-throughput screening of Mpro inhibitors using a robust fluorescence polarization (FP) assay. Candidate Mpro inhibitors from large compound libraries could be rapidly identified by monitoring the change of millipolarization unit value. This affordable FP assay can be modified to screen antiviral agents targeting virus protease. For complete details on the use and execution of this protocol, please refer to Li et al. (2022) , Yan et al. (2021) , and Yan et al. (2022c) .
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INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
·
2026
·
IF 8.7
Fangjun Wu,Juan Sun,Zheng Nie,Fujie Lai,Xudong Yang,Shaoyang Chen,Yang Wang
In the context of the ongoing normalisation of the SARS-CoV-2, the number of patients infected with the new coronavirus remains high on an annual basis. It is therefore imperative that individuals have access to rapid and cost-effective diagnostic tests. Conventional diagnostic tests,such as Reverse Transcription-Quantitative Polymerase Chain Reaction(RT-QPCR), Enzyme-Linked Immunosorbent Assay(ELISA) are costly and cumbersome, impeding their application in single-sample detection. This study has developed a cost-effective lateral flow immunoassay that rivals the sensitivity and simplicity of ELISA and colloidal gold tests, facilitating rapid and affordable detection of SARS-CoV-2 antigen. The synthesis of Fe 3 O 4 particles with a diameter of approximately 150 nm was initially conducted via a solvothermal method. Subsequently, the preparation of Fe 3 O 4 @PMAA particles with carboxyl groups, was undertaken through a reflux precipitation polymerization. Finally, the above particles were coupled with mouse anti-coronavirus SARS-CoV-2 N protein IgG by standard NHS/EDC methods to obtain Fe 3 O 4 @PMAA@IgG nanoprobe. This nanoprobe was then used as an immunoconjugate pad for the detection of the SARS-CoV-2 nucleoprotein. The nanoprobe-based method demonstrated significant advantages in cost-effectiveness and timeliness (with an assay duration of only 1 min), costing approximately $0.13 per sample. When used for SARS-CoV-2 antigen detection, this cost represents only 0.02% of that of RT-QPCR and 0.05% of that of ELISA.The detection limit of the lateral flow immunoassay is (0. 5 ng/mL).
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