CHEMBIOCHEM
·
2024
·
IF 2.6
Chunyu Yan,Wei Zhou,Yan Zhang,Xuelian Zhou,Qinglong Qiao,Lu Miao,Zhaochao Xu
In this study, super-resolution structured illumination microscope (SIM) was used to analyze molecular mechanism of endocytic acidification inhibitors in the SARS-CoV-2 pandemic, such as Chloroquine (CQ), Hydroxychloroquine (HCQ) and Bafilomycin A1 (BafA1). We fluorescently labeled the SARS-CoV-2 RBD and its receptor ACE2 protein with small molecule dyes. Utilizing SIM imaging, the real-time impact of inhibitors (BafA1, CQ, HCQ, Dynasore) on the RBD-ACE2 endocytotic process was dynamically tracked in living cells. Initially, the protein activity of RBD and ACE2 was ensured after being labeled. And then our findings revealed that these inhibitors could inhibit the internalization and degradation of RBD-ACE2 to varying degrees. Among them, 100 nM BafA1 exhibited the most satisfactory endocytotic inhibition (~63.9%) and protein degradation inhibition (~97.7%). And it could inhibit the fusion between endocytic vesicles in the living cells. Additionally, Dynasore, a widely recognized dynein inhibitor, also demonstrated cell acidification inhibition effects. Together, these inhibitors collectively hinder SARS-CoV-2 infection by inhibiting both the viral internalization and RNA release. The comprehensive evaluation of pharmacological mechanisms through super-resolution fluorescence imaging has laid a crucial theoretical foundation for the development of potential drugs to treat COVID-19.
研究领域:
病毒学
·
细胞生物学
·
药理学
·
成像科学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2021
·
IF 7.46
Yongqiang Li,Peixiang Ma,Quan Tao,Hans-Joachim Krause,Siwei Yang,Guqiao Ding,Hui Dong,Xiaoming Xie
The rapid and sensitive diagnosis of the highly contagious severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is one of the crucial issues at the outbreak of the ongoing global pandemic that has no valid cure. Here, we propose a SARS-CoV-2 antibody conjugated magnetic graphene quantum dots (GQDs)-based magnetic relaxation switch (MRSw) that specifically recognizes the SARS-CoV-2. The probe of MRSw can be directly mixed with the test sample in a fully sealed vial without sample pretreatment, which largely reduces the testers’ risk of infection during the operation. The closed-tube one-step strategy to detect SARS-CoV-2 is developed with home-made ultra-low field nuclear magnetic resonance (ULF NMR) relaxometry working at 118 μT. The magnetic GQDs-based probe shows ultra-high sensitivity in the detection of SARS-CoV-2 due to its high magnetic relaxivity, and the limit of detection is optimized to 248 Particles mL ‒1 . Meanwhile, the detection time in ULF NMR system is only 2 min, which can significantly improve the efficiency of detection. In short, the magnetic GQDs-based MRSw coupled with ULF NMR can realize a rapid, safe, and sensitive detection of SARS-CoV-2.
研究领域:
生物医学工程
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病毒学
·
分子诊断
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2022
·
IF 12.545
Yang Sun,Cheng Yang,Xiaolin Jiang,Pengbo Zhang,Shuo Chen,Fengxia Su,Hui Wang,Weiliang Liu,Xiaofei He,Lei Chen,Baoyuan Man,Zhengping Li
False detection of SARS-CoV-2 is detrimental to epidemic prevention and control. The scalar nature of the detected signal and the imperfect target recognition property of developed methods are the root causes of generating false signals. Here, we reported a collaborative system of CRISPR-Cas13a coupling with the stabilized graphene field-effect transistor, providing high-intensity vector signals for detecting SARS-CoV-2. In this collaborative system, SARS-CoV-2 RNA generates a “big subtraction” signal with a right-shifted feature, whereas any untargets cause the left-shifted characteristic signal. Thus, the false detection of SARS-CoV-2 is eliminated. High sensitivity with 0.15 copies/μL was obtained. In addition, the wide concerned instability of the graphene field-effect transistor for biosensing in solution environment was solved by the hydrophobic treatment to its substrate, which should be a milestone in advancing it's engineering application. This collaborative system characterized by the high-intensity vector signal and amazing stability significantly advances the accurate SARS-CoV-2 detection from the aspect of signal nature.
研究领域:
生物传感
·
纳米技术
·
分子生物学
·
电子学
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产品关联
DOI
PubMed
高影响因子
Journal of Biophotonics
·
2023
·
IF 2.8
Shuxia Lai,Yi Liu,Shubin Fang,Qiong Wu,Min Fan,Duo Lin,Jizhen Lin,Shangyuan Feng
The fast spread and transmission of the coronavirus 2019 (COVID-19) has become one of serious global public health problems. Herein, a surface enhanced Raman spectroscopy-based lateral flow immunoassay (LFA) was developed for the detection of SARS-CoV-2 antigen. Using uniquely designed core–shell nanoparticle with embedded Raman probe molecules as the indicator to reveal the concentration of target protein, excellent quantitative performance with a limit of detection (LOD) of 0.03 ng/mL and detection range of 10–1000 ng/mL can be achieved within 15 min. Besides, the detection of spiked virus protein in human saliva was also performed with a portable Raman spectrometer, proposing the feasibility of the method in practical applications. This easy-to-use, rapid and accurate method would provide a point-of-care testing way as the ideal alternative for current detection requirement of virus-related biomarkers.
研究领域:
分析化学
·
生物医学工程
·
病毒学
·
公共卫生
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产品关联
DOI
PubMed
高影响因子
APPLIED SURFACE SCIENCE
·
2024
·
IF 6.3
Jialin Zhao,Ting Wu,Youlin Xiong,Wei Zhang,Zian Pan,Shuangfei Cai,Rong Yang
Electrochemical biosensors have shown promise in bioanalysis and diagnosis. However, they suffer from the accumulation and adhesion of biomolecules in biofluids on sensing surfaces, while the hiring of potential antifouling is limited by the cost and complexity of the construction methods. Here, an economical three-dimensional (3D) conductive antifouling coating was prepared. This coating was composed of bovine serum albumin (BSA) crosslinked glutaraldehyde (GA) and copper-gold nanowires (CuAu NWs) with excellent electrical conductivity. Furthermore, a dual-targeted aptamer (D-Apt), with a “3-in-1″ structure of two specific binding sequences and a DNA linker, was designed to specifically recognize BSA and the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike (S) protein, affording low nanomolar binding affinity. Importantly, the introduction of the D-Apt greatly simplified the functionalization procedure on the electrode surface and the immobilization of the biorecognition element. Then, a simple electrochemical biosensor based on the CuAu/BSA/GA and D-Apt was constructed for the detection of S protein. This sensor exhibited merits of cost-effectiveness, sensitivity and practicality for detection of S protein. The work could provide a facile avenue to construct electrochemical biosensors with low-cost antifouling for the accurate diagnosis of epidemic antigenic disease markers.
研究领域:
生物医学工程
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分析化学
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病毒学
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产品关联
DOI
高影响因子
ARCHIV DER PHARMAZIE
·
2025
·Vol. 358
·期 9
·e70089
·
IF 3.6
Leyao Chen,Xinluan Lv,Xiaoyu Chang,Ruiyong Wang
The SARS-CoV-2 pandemic has spurred global efforts to develop therapeutic approaches. The main protease (Mpro) of SARS-CoV-2 is crucial for viral replication and a key target for therapeutic development. In this study, 22 thiosemicarbazone derivatives were synthesized. Enzyme activity assays showed compound 3c exhibited obvious inhibition of Mpro, with an IC 50 value of 3.89 μM. The interaction mechanisms between thiosemicarbazone derivatives and Mpro were investigated using synchronous fluorescence, three-dimensional fluorescence, and circular dichroism spectroscopy. The fluorescence results indicate that static quenching is predominant. The equilibrium dissociation constant ( K d ) of 2.3 μM between 3c and Mpro was obtained by microscale thermophoresis. This K d value demonstrates the binding affinity of 3c for Mpro, consistent with the spectral results. Meanwhile, the binding modes and stability of thiosemicarbazone derivatives with Mpro were evaluated through molecular docking and molecular dynamics simulations, which also confirmed the stable binding between compound 3c and Mpro. This study provides valuable insights into the interaction mechanism between thiosemicarbazone derivatives and Mpro and provides clues for the design of Mpro inhibitors.
研究领域:
病毒学
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药物化学
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结构生物学
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分子药理学
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抗病毒治疗
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生物物理化学
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计算生物学
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产品关联
DOI
PubMed
高影响因子
TALANTA
·
2021
·
IF 6.057
Chenchen Ge,Juan Feng,Jiaming Zhang,Kai Hu,Dou Wang,Ling Zha,Xuejuan Hu,Rongsong Li
Nucleocapsid protein (N protein) is the most abundant protein in SARS-CoV2 and is highly conserved, and there are no homologous proteins in the human body, making it an ideal biomarker for the early diagnosis of SARS-CoV2. However, early detection of clinical specimens for SARS-CoV2 remains a challenge due to false-negative results with viral RNA and host antibodies based testing. In this manuscript, a microfluidic chip with femtoliter-sized wells was fabricated for the sensitive digital detection of N protein. Briefly, β-galactosidase (β-Gal)-linked antibody/N protein/aptamer immunocomplexes were formed on magnetic beads (MBs). Afterwards, the MBs and β-Gal substrate fluorescein-di-β- d -galactopyranoside (FDG) were injected into the chip together. Each well of the chip would only hold one MB as confined by the diameter of the wells. The MBs in the wells were sealed by fluorocarbon oil, which confines the fluorescent (FL) product generated from the reaction between β-Gal and FDG in the individual femtoliter-sized well and creates a locally high concentration of the FL product. The FL images of the wells were acquired using a conventional inverted FL microscope. The number of FL wells with MBs (FL wells number) and the number of wells with MBs (MBs wells number) were counted, respectively. The percentage of FL wells was calculated by dividing (FL wells number) by (MBs wells number). The higher the percentage of FL wells, the higher the N protein concentration. The detection limit of this digital method for N protein was 33.28 pg/mL, which was 300 times lower than traditional double-antibody sandwich based enzyme-linked immunosorbent assay (ELISA).
研究领域:
生物化学
·
分子生物学
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生物医学工程
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病毒学
·
分析化学
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产品关联
DOI
PubMed
高影响因子
Small
·
2023
·Vol. 19
·期 20
·2208167
·
IF 13.3
Mengya He,Xican Xu,Hongyu Wang,Qingyuan Wu,Linghao Zhang,Dongsheng Zhou,Yigang Tong,Xin Su,Huiyu Liu
Fluorescence-based PCR and other amplification methods have been used for SARS-CoV-2 diagnostics, however, it requires costly fluorescence detectors and probes limiting deploying large-scale screening. Here, a cut-price colorimetric method for SARS-CoV-2 RNA detection by iron manganese silicate nanozyme (IMSN) is established. IMSN catalyzes the oxidation of chromogenic substrates by its peroxidase (POD)-like activity, which is effectively inhibited by pyrophosphate ions (PPi). Due to the large number of PPi generated by amplification processes, SARS-CoV-2 RNA can be detected by a colorimetric readout visible to the naked eye, with the detection limit of 240 copies mL −1 . This conceptually new method has been successfully applied to correctly distinguish positive and negative oropharyngeal swab samples of COVID-19. Colorimetric assay provides a low-cost and instrumental-free solution for nucleic acid detection, which holds great potential for facilitating virus surveillance.
研究领域:
生物技术
·
分子生物学
·
病毒学
·
诊断方法
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MEDICINAL CHEMISTRY
·
2024
·
IF 6.8
Yahong Tan,Jinyue Yang,Min Wang,Qi Peng,Yongqi Li,Lifeng Fu,Mengmeng Zhang,Jiang Wu,Guanya Yang,Christopher John Hipolito,Youming Zhang,Jianxun Qi,Yi Shi,Yizhen Yin
Macrocyclic peptides have garnered significant attention as promising drug candidates. However, they typically face challenges in achieving and enhancing cell permeability for access to intracellular targets. In this study, we focused on the de novo screening of macrocyclic peptide inhibitors against the main protease (Mpro) of SARS-CoV-2 and identified novel noncovalently bound macrocyclic peptides that effectively inhibit proteolytic activity. High-resolution crystal structures further revealed molecular interactions between the macrocyclic peptides and Mpro. Subsequently, a specific macrocyclic peptide lacking cell permeability was further optimized and transformed into a low-toxicity, metabolically stable bicyclic peptide with a cell penetration capacity and therapeutic potential against SARS-CoV-2. The bicyclic peptide was achieved using a novel strategy that involved introducing both a bicyclic structure and a bridging perfluorobiphenyl group. Our study not only provides a lead peptide inhibitor for COVID-19 but also offers valuable insights into achieving cell penetration for macrocyclic peptides through strategic modifications.
研究领域:
药理学
·
病毒学
·
药物发现
·
蛋白质工程
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DOI
高影响因子
CELL
·
2022
·
IF 66.85
Liming Yan,Yucen Huang,Ji Ge,Zhenyu Liu,Pengchi Lu,Bo Huang,Shan Gao,Junbo Wang,Liping Tan,Sihan Ye,Fengxi Yu,Weiqi Lan,Shiya Xu,Feng Zhou,Lei Shi,Luke W. Guddat,Yan Gao,Zihe Rao,Zhiyong Lou
Summary Decoration of cap on viral RNA plays essential roles in SARS-CoV-2 proliferation. Here, we report a mechanism for SARS-CoV-2 RNA capping and document structural details at atomic resolution. The NiRAN domain in polymerase catalyzes the covalent link of RNA 5′ end to the first residue of nsp9 (termed as RNAylation), thus being an intermediate to form cap core (GpppA) with GTP catalyzed again by NiRAN. We also reveal that triphosphorylated nucleotide analog inhibitors can be bonded to nsp9 and fit into a previously unknown “Nuc-pocket” in NiRAN, thus inhibiting nsp9 RNAylation and formation of GpppA. S-loop (residues 50-KTN-52) in NiRAN presents a remarkable conformational shift observed in RTC bound with sofosbuvir monophosphate, reasoning an “induce-and-lock” mechanism to design inhibitors. These findings not only improve the understanding of SARS-CoV-2 RNA capping and the mode of action of NAIs but also provide a strategy to design antiviral drugs.
研究领域:
病毒学
·
分子生物学
·
结构生物学
·
药物设计
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产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2024
·
IF 10.7
Chunxia Chen,Shujie Duan,Jianing Ji,Mengli Wu,Zhan Yang,Mengyang Cai,Menghan Xue,Lanju Wang,Rui Chen,Sima Yaron,Keying Guo,Sergio Benardini,Zhizeng Wang,Yang Luo
Conventional point-of-care lateral flow immunoassays are characterized by an antibody-tagged probe irregular coupling that can limit sensitivity and require a long assay’s time. We synthesized polyethylene glycol-modified selenium nanoparticles (PEG-SeNPs) by template method and developed a strategy to set antibody probes targeted and orderly by using PEG-SeNPs. Synthesized PEG-SeNPs with high stability could immobilize antibodies in the “stand-up” orientation, resulting in a faster detection time of less than 1 min by direct observer visualization without instruments or equipment. Results showed that SARS-CoV-2 antibody could be systematically structured on the chip, resulting in a detection limit of 10 pg/mL, significantly higher than conventional chips. The new device has been validated on 192 clinical samples and we found 100% negative coincidence, 93.94% positive coincidence, and 95.83% overall coincidence with reverse transcriptional PCR test. The orderly arrayed probe’s stability allowed to detect throat swabs, saliva, serum, fingertip blood samples, and mutant strains without cross-reactivity with common respiratory viruses or pathogenic strains, demonstrating promising potential for a universal colorimetric platform for ultrafast field-deployable diagnostics.
研究领域:
生物医学工程
·
纳米技术
·
病毒学
·
诊断医学
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产品关联
DOI
PubMed
高影响因子
Cell Discovery
·
2023
·Vol. 9
·期 1
·1-15
·
IF 33.5
Ai Liangxia,Li Yafei,Zhou Li,Yao Wenrong,Zhang Hao,Hu Zhaoyu,Han Jinyu,Wang Weijie,Wu Junmiao,Xu Pan,Wang Ruiyue,Li Zhangyi,Li Zhouwang,Wei Chengliang,Liang Jianqun,Chen Haobo,Yang Zhimiao,Guo Ming,Huang Zhixiang,Wang Xin,Zhang Zhen,Xiang Wenjie,Sun Dazheng,Xu Lianqiang,Huang Meiyan,Lv Bin,Peng Peiqi,Zhang Shangfeng,Ji Xuhao,Luo Huiyi,Chen Nanping,Chen Jianping,Lan Ke,Hu Yong
Advanced mRNA vaccines play vital roles against SARS-CoV-2. However, most current mRNA delivery platforms need to be stored at −20 °C or −70 °C due to their poor stability, which severely restricts their availability. Herein, we develop a lyophilization technique to prepare SARS-CoV-2 mRNA-lipid nanoparticle vaccines with long-term thermostability. The physiochemical properties and bioactivities of lyophilized vaccines showed no change at 25 °C over 6 months, and the lyophilized SARS-CoV-2 mRNA vaccines could elicit potent humoral and cellular immunity whether in mice, rabbits, or rhesus macaques. Furthermore, in the human trial, administration of lyophilized Omicron mRNA vaccine as a booster shot also engendered strong immunity without severe adverse events, where the titers of neutralizing antibodies against Omicron BA.1/BA.2/BA.4 were increased by at least 253-fold after a booster shot following two doses of the commercial inactivated vaccine, CoronaVac. This lyophilization platform overcomes the instability of mRNA vaccines without affecting their bioactivity and significantly improves their accessibility, particularly in remote regions.
研究领域:
病毒学
·
免疫学
·
药学
·
生物技术
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产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）