VIROLOGY
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2022
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IF 3.513
Haohao Yan,Zhicheng Liu,Gangan Yan,Xiaoli Liu,Xiaoping Liu,Yanchang Wang,Yunyu Chen
The global scourge of COVID-19 is a serious threat to public health, but effective therapies remain very limited for this disease. Therefore, the discovery of novel antiviral agents is urgently needed to fight against COVID-19. In the lifecycle of SARS-CoV-2, the causing pathogen of COVID-19, papain-like protease (PLpro) is responsible for the cleavage of polyprotein into functional units as well as immune evasion of vaccines. Hence, PLpro has been regarded as an attractive target to develop antiviral agents. Herein, we first developed a robust and simple sandwich-like fluorescence polarization (FP) screening assay for the discovery of PLpro inhibitors, and identified anacardic acid as a novel competitive inhibitor against PLpro in vitro with an IC 50 value of 24.26 ± 0.4 μM. This reliable FP screening assay could provide a prospective avenue for rapid discovery of antiviral agents targeting PLpro in a large-scale screening.
研究领域:
病毒学
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药理学
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分子生物学
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Infectious Diseases and Clinical Microbiology
·
2026
·Vol. 8
·期 2
·165
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IF 1.1
Jingping Yang,Dan Liu,Xiyuan Xu,Na Meiri,Dezhi Yang
Objective This study aimed to develop a sensitive and reliable method for detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in various environmental samples using nano–liquid chromatography–tandem mass spectrometry parallel reaction monitoring (nanoLC-MS/MS PRM), targeting sequences of the viral spike (S) and nucleocapsid (N) proteins. Materials and Methods A synthetic peptide screening approach was employed to systematically evaluate candidate sequences as surrogates for protein quantification. The process began with the digestion of purified SARS-CoV-2 proteins. The candidate sequences were then refined to a subset of target peptides, and stable isotope-labeled heavy peptides were used as standards in a PRM workflow. The detection limits were determined using the nanoLC-MS/MS PRM method. Results The developed method achieved limits of detection in the 1 fmol/µL (2–8 × 10-15 viral copies/L) range on-column. This demonstrates high sensitivity for detecting SARS-CoV-2 in environmental samples using the PRM method. Conclusion The study successfully developed a sensitive and reliable peptide probe-based approach for detecting SARS-CoV-2 in various environmental samples. The use of the PRM method targeting the S and N proteins of SARS-CoV-2 provides a robust approach for virus detection, particularly given the potential persistence of the virus on surfaces. Graphic Graphic . Open in a new tab
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Advanced Science
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2026
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IF 14.1
Sidi Yang,Kun Li,Lihong Liu,Linsen Zeng,Qifeng Deng,Jiacheng Huang,Xiaoran Dong,Xin Wang,Jianwei Liang,Hongchao Liu,Hong Peng,Yuxin Lin,Xiaolu Xie,Yuzhen Ye,Tiefeng Xu,Zhaohuan Wang,Chun-Mei Li,Deyin Guo
SARS-CoV-2 accessory protein ORF3a contributes to viral pathogenesis through membrane remodeling, immune evasion, and inflammation induction. However, the molecular mechanisms underlying ORF3a-mediated pathogenesis remain poorly characterized, and no therapeutic strategies targeting ORF3a currently exist. Here, we demonstrate that palmitoylation, a post-translational modification, governs ORF3a-mediated viral pathogenesis. Specifically, ORF3a undergoes ZDHHC18-mediated palmitoylation at evolutionarily conserved Cys130/Cys133 residues, which stabilizes the protein by masking an intrinsic proteasomal degradation signal. This palmitoylation competitively inhibits tripartite motif-containing 16 (TRIM16)-dependent K27-linked polyubiquitination, thereby preventing ORF3a degradation and enhancing viral replication and inflammatory responses. A designed ORF3a-mimicking palmitoylation-inhibitory peptide (OPIP) blocked ORF3a palmitoylation, promoted its degradation, and significantly reduced SARS-CoV-2 pathogenicity. Collectively, these findings establish ZDHHC18-mediated palmitoylation as a central regulator of ORF3a stability and virulence, revealing a potentially druggable axis for disrupting SARS-CoV-2 pathogenesis.
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MICROCHEMICAL JOURNAL
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2026
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IF 5.2
Dandan Chen,Bin Yin,Zhirun Ji,Zhiqi Liu,Jiahao Li,Fei Liu,Xuping Wu
Amplification-free digital detection of SARS-CoV-2 via CRISPR/Cas12a. • Pneumatically actuated droplet microfluidics enables uniform ∼30 μm droplets. • Rapid detection of SARS-CoV-2 with demonstrated performance in clinical samples. • Offers a simple, portable method for rapid virus detection. Outbreaks of respiratory viruses, including severe acute respiratory syndrome coronavirus 2(SARS-CoV-2), Influenza A (IAV), and Influenza B (IBV) viruses, present a formidable challenge to global public health, driving the urgent need for rapid, sensitive, and deployable diagnostic tools. Here, we present an amplification-free digital nucleic acid detection platform that synergistically integrates a pneumatically actuated droplet microfluidics platform (PADMP) with CRISPR/Cas12a. PADMP enables the precise generation of highly uniform, monodisperse microdroplets (∼30 μm), within which an optimized Cas12a reaction is compartmentalized. This digital partitioning strategy exponentially increases the local target concentration and facilitates single-molecule detection without pre-amplification. Through systematic optimization of droplet stability, surfactant formulation, and Cas12a assay kinetics, the PADMP-Cas12a platform achieves a detection limit (LOD) of 100 pM for synthetic SARS-CoV-2 N gene targets within 40 min. It demonstrates exceptional specificity, showing no cross-reactivity with non-target respiratory viruses (IAV, IBV, SARS-CoV, MERS-CoV, HRSV). The platform successfully detected SARS-CoV-2 in clinical positive samples, with mean droplet fluorescence intensity negatively correlated with Ct values. By combining the advantages of droplet microfluidics with the programmable recognition and trans -cleavage activity of CRISPR/Cas12a, this platform provides a rapid, amplification-free, and highly specific digital detection pathway for RNA viruses, marking an important advance toward high-throughput and point-of-care testing (POCT) for molecular diagnostics. Download: Download high-res image (127KB) Download: Download full-size image
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Journal of Materials Chemistry B
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2026
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IF 6.2
Xiaoyu Ma,Zhuang Zhang,Dandan Yang,Hui Wang,Hao Zhang,Yue Yang,Siying Li,Yuping Shan
Beyond the canonical angiotensin-converting enzyme 2 (ACE2)-mediated entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into host cells, basigin (BSG) and dipeptidyl peptidase 4 (DPP4) may also act as alternative receptors that can facilitate viral attachment and subsequent endocytosis. Herein, we analyzed the binding efficiency and binding dynamics of SARS-CoV-2 virus-like particles (VLPs), mediated by BSG and DPP4 at the single-particle level, by combining super-resolution direct stochastic optical reconstruction microscopy (dSTORM) and single-molecule force spectroscopy (SMFS). dSTORM imaging demonstrated that DPP4 exhibits higher surface expression levels than BSG on HepG2 cells. However, SARS-CoV-2 VLPs display a distinct preference for BSG-dependent cellular entry. Using SMFS, we further probed the binding dynamics between the SARS-CoV-2 spike (S) protein receptor-binding domain (RBD) and these two receptors. The results demonstrated that RBD-BSG exhibits greater stability and a more rapid formation rate compared to RBD-DPP4. Subsequently, a real-time force tracing technique was used to show that BSG-mediated VLP entry requires lower force and faster speed compared with DPP4-mediated entry. In particular, we confirmed that the structural integrity of intramolecular disulfide bonds within the RBD is indispensable for mediating high-affinity receptor binding. These findings elucidate the distinct receptor-mediated endocytic dynamic mechanisms underlying SARS-CoV-2 entry mediated by BSG and DPP4, expanding our understanding of viral invasion beyond the ACE2 pathway and providing insights into potential intervention strategies targeting viral infection.
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JOURNAL OF PROTEOME RESEARCH
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2026
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IF 3.8
Xue Li,Zhanxuan Emily Wu,Xinyuan Wang,Fengming Luo,Cheng Deng,Yu Chen,Huajing Wan,Xiaoju Tang,Yabin Zhang,Peiliang Shi,Zhengzhong Wu,Chunyou Ning,Wenxiang Yang,Hao Yang,Jia Liu,Fei Li
Age is a key risk factor for morbidity in coronavirus disease 2019 (COVID-19). This study investigates how “inflammaging” predisposes older hosts to severe SARS-CoV-2 outcomes through systemic and localized metabolic shifts. SARS-CoV-2 infection experiments were conducted respectively on 8 week-old mice and 16 week-old mice. An integrated multiomics approach was employed to delineate age-dependent molecular signatures and metabolic-immune crosstalk. 16 week-old mice exhibited significantly exacerbated weight loss and pulmonary pathology, successfully recapitulating age-related susceptibility. Multiomics integration revealed a “systemic bioenergetic gap”: suppressed plasma TCA cycle metabolites correlated with depleted pulmonary nucleotide pools (AMP, GMP, UMP and uracil). This metabolic failure led to a paradoxical proteomic profile: despite interferon pathway activation, key antiviral effectors (RSAD2, ISG15, IFIT3B) were downregulated, while stress markers (CYP1A1, MAPK8) increased. Furthermore, threonic acid, betaine, and d-ribose were identified as robust, age-dependent biomarkers of infection severity. Our findings suggest that COVID-19 severity in the aging host is driven by the failure of a systemic-to-local metabolic-immune axis. The exhaustion of circulating energetic precursors constrains localized pulmonary nucleotide metabolism, thereby impairing essential antiviral responses and tissue repair. This study identifies the “systemic bioenergetic gap” as a novel therapeutic target, suggesting that systemic metabolic resuscitation may improve clinical outcomes in elderly populations.
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BIOORGANIC CHEMISTRY
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2021
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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.
研究领域:
病毒学
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药学
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生物技术
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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.
研究领域:
生物传感器
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分析化学
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材料科学
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病毒学
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生物医学工程
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BIOORGANIC & MEDICINAL CHEMISTRY
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2022
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IF 3.461
Ting Xu,Jie-Ru Meng,Wanqing Cheng,Jia-Zheng Liu,Junyan Chu,Qian Zhang,Nannan Ma,Li-Ping Bai,Yong Guo
Honokiol, isolated from a traditional Chinese medicine (TCM) Magnolia officinalis , is a biphenolic compound with several biological activities. To improve and broaden its biological activity, herein, two series of honokiol thioethers bearing 1,3,4-oxadiazole moieties were prepared and assessed for their α -glucosidase and SARS-CoV-2 entry inhibitory activities. Among all the honokiol thioethers, compound 7l exhibited the strongest α -glucosidase inhibitory effect with an IC 50 value of 18.9 ± 2.3 µM, which was superior to the reference drug acarbose (IC 50 = 24.4 ± 0.3 µM). Some interesting results of structure–activity relationships (SARs) have also been discussed. Enzyme kinetic study demonstrated that 7l was a noncompetitive α -glucosidase inhibitor, which was further supported by the results of molecular docking. Moreover, honokiol thioethers 7e , 9a , 9e , and 9r exhibited potent antiviral activity against SARS-CoV-2 pseudovirus entering into HEK-293 T-ACE2 h . Especially 9a displayed the strongest inhibitory activity against SARS-CoV-2 pseudovirus entry with an IC 50 value of 16.96 ± 2.45 μM, which was lower than the positive control Evans blue (21.98 ± 1.98 μM). Biolayer interferometry (BLI) binding and docking studies suggested that 9a and 9r may effectively block the binding of SARS-CoV-2 to the host ACE2 receptor through dual recognition of SARS-CoV-2 spike RBD and human ACE2. Additionally, the potent honokiol thioethers 7l , 9a , and 9r displayed relatively no cytotoxicity to normal cells (LO2). These findings will provide a theoretical basis for the discovery of honokiol derivatives as potential both α -glucosidase and SARS-CoV-2 entry inhibitors.
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BIOSENSORS & BIOELECTRONICS
·
2023
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IF 12.6
Jiareng Chen,Willis Kwun Hei Ho,Bohan Yin,Qin Zhang,Chuanqi Li,Jiaxiang Yan,Yingying Huang,Jianhua Hao,Changqing Yi,Yu Zhang,Siu Hong Dexter Wong,Mo Yang
Upconversion nanoparticles (UCNPs) are ideal donors for luminescence resonance energy transfer (LRET)-based biosensors due to their excellent upconversion luminescence properties . However, the relatively large size of antibodies and proteins limits the application of UCNPs-based LRET biosensors in protein detection because the large steric hindrance of proteins leads to low energy transfer efficiency between UCNPs and receptors. Herein, we developed a magnetic responsive UCNPs-based LRET biosensor to control the coupling distance between antibody-functionalized UCNPs (Ab-UCNPs) as donors and antibody-PEG linker-magnetic gold nanoparticles (Ab-PEG-MGNs) as acceptors for ultrasensitive and highly selective detection of SARS-CoV-2 spike proteins . Our results showed that this platform reversibly shortened the coupling distance between UCNPs and MGNs and enhanced the LRET signal with a 10-fold increase in the limit of detection (LOD) from 20.6 pg/mL without magnetic modulation to 2.1 pg/mL with magnetic modulation within 1 h. The finite-difference time-domain (FDTD) simulation with cyclic distance change confirmed the distance-dependent LRET efficiency under magnetic modulation, which supported the experimental results. Moreover, the applications of this magnetic-responsive UCNP-based LRET biosensor could be extended to other large-size biomolecule detection.
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SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY
·
2023
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IF 4.4
Yansheng Liu,Zhenle Qin,Xiaobo Jia,Jin Zhou,Hongli Li,Xiaohong Wang,Yating Chen,Junpeng Deng,Zhicheng Jin,Guofu Wang
The global pandemic of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a great disaster to the economy and human society. Nowadays, SARS-CoV-2 is fading away from people’s memory but it still exists around us. PCR plays an important role in detecting SARS-CoV-2 but it requires a long detecting time, equipped laboratory, and professional operators. In comparison with polymerase chain reaction (PCR), surface-enhanced Raman scattering (SERS) is a promising method for detecting SARS-CoV-2 due to its fast, easily operated, and high-sensitivity properties. In this study, the monolayer Ag nanoparticles (MAgNPs) covered with single-layer graphene (SLG) are applied as a SERS substrate. The angiotensin converting enzyme 2 (ACE2) is selected as a bio-probes that can specifically bind to the SARS-CoV-2 S protein. The SERS-based biosensor is formed by ACE2 functionalized SLG/MAgNPs and the LODs of detecting SARS-CoV-2 S protein in phosphate-buffered saline (PBS) and in pharyngeal swabs solution (PSS) are 0.1 fg mL −1 and 10 fg mL −1 , respectively. This biosensor provides a way of directly detecting SARS-CoV-2 S protein with high sensitivity and specificity. It illustrates a practical potential in the rapid detection of the SARS-CoV-2 virus.
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ACS Sensors
·
2023
·
IF 8.9
Sihan Wang,Yougang Zhong,Zhen Gong,Xiaoli Zhu,Kai Wen,Shuhua Wei,Zhiwei He,Zile Wang,Jincheng Xiong,Shuai Zhang,Xiaotian Liu,Liang Zhang,Jianzhong Shen,Haiyang Jiang
Waves of COVID-19 outbreaks have dragged down the global economy and endangered human life. There is an urgent need for timeliness and sensitive SARS-CoV-2 detection techniques to complement the existing PCR assay. Herein, the controllable growth of gold crystalline grains was achieved by applying the reverse current during pulse electrochemical deposition (PED) interval. The proposed method validates the effects of pulse reverse current (PRC) on the atomic arrangement, crystal structures, orientations, and film characteristics in Au PED. The gap between the gold grains on the surface of the nanocrystalline gold interdigitated microelectrodes (NG-IDME) fabricated by the PED+PRC process matches the size of the antiviral antibody. Immunosensors are prepared by binding a large number of antiviral antibodies on the surface of NG-IDME. The NG-IDME immunosensor has a high specific capture ability for SARS-CoV-2 nucleocapsid protein (SARS-CoV-2/N-Pro) and completes ultrasensitive and quantification of SARS-CoV-2/N-Pro in humans and pets within 5 min (the LOQ as low as 75 fg/mL). The specificity, accuracy, stability, and actual blind sample tests show that the NG-IDME immunosensor is suitable for the detection of SARS-CoV-2 in humans and animals. This approach assists in monitoring the transmission of SARS-CoV-2-infected animals to humans.
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