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JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES · 2024 · IF 2.8

A validated LC-MS/MS method for determination of six Anti-SARS-CoV-2 drugs in plasma and its application for a pharmacokinetic study in rats

Zong-liang Xu,Cheng-jian Li,Xian Qian,Hu Duan,Jin Zhou,Qian-qian Zhang,Xin Dong,Liang Zhao

Antiviral treatment for COVID-19 is considered an effective tool in reducing the rate of severe cases and deaths. As of June 2023, a total of six small molecule antiviral drugs have been conditionally approved for marketing by the National Medical Products Administration (NMPA) within China. In this study, a method of HPLC-MS/MS was established and validated for the determination of six small molecule antiviral drugs in plasma using Lamivudine as an internal standard. The chromatographic separation was performed using gradient elution with an ACE 3 C 18 -PFP column (3.0 mm × 150 mm, 3 μm), and the mobile phase consisted of deionized water and acetonitrile/water (90:10, v/v), both with 10 mmol/L of ammonium acetate and 0.1 % ammonium hydroxide added. Quantitative analysis of the six small molecule drugs was carried out through selective reaction monitoring based on the positive ion spray ionization mode. The method exhibited excellent precision, accuracy, recovery, and linearity, and it was used to determine the pharmacokinetic characteristics in rats. Our work not only established a bioanalytical method for six small molecule antiviral drugs but also provided scientific references for clinical pharmacokinetic studies.

研究领域: 药理学 · 分析化学
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BIOSENSORS & BIOELECTRONICS · 2024 · IF 10.7

One-step time-resolved cascade logic gate microfluidic chip for home testing of SARS-CoV-2 and flu B

Jingwei Chen,Tingting Liu,Yule Zhang,Mengnan Duan,Zhijin Yang,Mengya Chen,Yiran Wang,Lulu Zheng,Songlin Zhuang,Dawei Zhang

Home testing technology strategy is critical for early screening of disease. However, current home testing technologies often require complex processes, which limits their application. In this study, a time-resolved cascade logic gate microfluidic chip (TCLMC) was revealed to enable capillary force-based one-step operation without manual intervention or professional equipment. By analogy with logic gates in the circuit, TCLMC could automatically control the fluid flow and regulate the incubation time to optimize the immunoassay . The limit of detection of TCLMC for SARS-CoV-2 and influenza B virus (Flu B) was 134.94 and 79.17 pg mL −1 within 10 min. Additionally, this study tested saliva samples from 12 Flu B patients and 24 healthy controls to verify its clinical application. The results showed that TCLMC had high sensitivity (100%), specificity (100%), and accuracy (100%). This study provides a new one-step strategy for home testing and demonstrates its great potential in the diagnosis field.

研究领域: 医学诊断 · 生物技术 · 微流体技术
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Algal Research-Biomass Biofuels and Bioproducts · 2025 · IF 4.5

SARS-CoV-2 Mpro and its inhibitory peptides from Spirulina platensis: Preparation, identification, in vitro and in silico characterization of activity and binding

Qiao-Hui Zeng,Zhi-En Yao,Xian-Min Zhou,Ping Qi,Xin-An Zeng

Inhibition of the biological activity of SARS-CoV-2 main protease (Mpro) effectively prevents viral replication, thereby presenting a promising therapeutic target for Corona Virus Disease 2019 (COVID-19) intervention. This study aims to screen the Mpro inhibitory peptides from the natural nutritional supplement Spirulina platensis ( S. platensis ). Firstly, Mpro was successfully prepared by heterologous expression in E. coli and Mpro's activity was detected. Meanwhile, four distinct peptides derived from S. platensis were obtained through enzymatic hydrolysis of protein. Subsequently, highly active peptides were synthesized and their inhibition effects on Mpro were verified by in vitro experiment and computer simulations. Results showed that the extraction rate of the protein from S. platensis was 96.11 %, the activity of Mpro was 845.90 U mg −1 , the peptide MQGPNY could inhibit the activity of Mpro at the inhibition rate of 20.21 % ± 3.8 % at the concentration of 2 mmol L −1 . Furthermore, results from computer Simulation showed that the peptide MQGPNY forms four pairs of hydrogen bonds with Mpro, which are Gln189, Ser46, Thr26 and Glu166. In addition, there were ten free residues involved in hydrophobic contacts, including His164, His41, Thr24, Met49, Thr45, Thr25, Gly143, Asn142, Cys145 and Met165. The foundings could provide theoretical support for the use of protein and its hydrolysates from S. platensis in functional food and supplement formulations in the post-epidemic era.

CAS / MDL: 79-11-8
研究领域: 生物化学 · 分子生物学 · 药理学
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STAR Protocols · 2022 · IF 0

Protocol for high-throughput screening of SARS-CoV-2 main protease inhibitors using a robust fluorescence polarization assay

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) .

研究领域: 病毒学 · 药学 · 分子生物学
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TALANTA · 2023 · IF 6.1

PdPtRu trimetallic nanozymes and application to electrochemical immunosensor for sensitive SARS-COV-2 antigen detection

Ying Li,Jian Xue,Rongjun Yu,Siling Chen,Xiangyu Deng,Anyi Chen,Jingfu Qiu

Herein, a ternary PdPtRu nanodendrite as novel trimetallic nanozyme was reported, which possessed excellent peroxidase-like activity as well as electro-catalytic activity on account of the synergistic effect between the three metals. Based on the excellent electro-catalytic activity of trimetallic PdPtRu nanozyme toward the reduction of H 2 O 2 , the trimetallic nanozyme was applied to construct a brief electrochemical immunosensor for SARS-COV-2 antigen detection. Concretely, trimetallic PdPtRu nanodendrite was used to modify electrode surface, which not only generated high reduction current of H 2 O 2 for signal amplification, but also provided massive active sites for capture antibody (Ab 1 ) immobilization to construct immunosensor. In the presence of target SARS-COV-2 antigen, SiO 2 nanosphere labeled detection antibody (Ab 2 ) composites were introduced on the electrode surface according sandwich immuno-reaction. Due to the inhibitory effect of SiO 2 nanosphere on the current signal, the current signal was decreased with the increasing target SARS-COV-2 antigen concentration. As a result, the proposed electrochemical immunosensor presented sensitive detection of SARS-COV-2 antigen with linear range from 1.0 pg/mL to 1.0 μg/mL and limit of detection down to 51.74 fg/mL. The proposed immunosensor provide a brief but sensitive antigen detection tool for rapid diagnosis of COVID-19.

CAS / MDL: 50-81-7 10049-07-7
研究领域: 生物医学工程 · 分析化学 · 病毒学
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Frontiers in Molecular Biosciences · 2022 · IF 6.113

Molecular Mechanism of SARS-CoVs Orf6 Targeting the Rae1–Nup98 Complex to Compete With mRNA Nuclear Export

Li Tinghan,Wen Yibo,Guo Hangtian,Yang Tingting,Yang Haitao,Ji Xiaoyun

The accessory protein Orf6 is uniquely expressed in sarbecoviruses including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which is an ongoing pandemic. SARS-CoV-2 Orf6 antagonizes host interferon signaling by inhibition of mRNA nuclear export through its interactions with the ribonucleic acid export 1 (Rae1)–nucleoporin 98 (Nup98) complex. Here, we confirmed the direct tight binding of Orf6 to the Rae1-Nup98 complex, which competitively inhibits RNA binding. We determined the crystal structures of both SARS-CoV-2 and SARS-CoV-1 Orf6 C-termini in complex with the Rae1–Nup98 heterodimer. In each structure, SARS-CoV Orf6 occupies the same potential mRNA-binding groove of the Rae1–Nup98 complex, comparable to the previously reported structures of other viral proteins complexed with Rae1-Nup98, indicating that the Rae1–Nup98 complex is a common target for different viruses to impair the nuclear export pathway. Structural analysis and biochemical studies highlight the critical role of the highly conserved methionine (M58) of SARS-CoVs Orf6. Altogether our data unravel a mechanistic understanding of SARS-CoVs Orf6 targeting the mRNA-binding site of the Rae1–Nup98 complex to compete with the nuclear export of host mRNA, which further emphasizes that Orf6 is a critical virulence factor of SARS-CoVs.

研究领域: 病毒学 · 分子生物学 · 结构生物学 · 细胞生物学
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ACS Applied Nano Materials · 2024 · IF 5.3

SERS/EC Dual-Mode Sensing System for the Assay of SARS-CoV-2 RNA by Using Single Gold Nanowires as a Platform

Qingshan Dai,Jiarui Huang,Xia Qiu,Xiaohu Chen,Yongxin Li

The continuous spread and infection of the severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) worldwide has brought a great impact on people’s lives and health and become a serious public health problem. In this contribution, a surface-enhanced Raman scattering/electrochemical (SERS/EC) dual-mode sensing system using single Au nanowires (Au NWs) as a platform is proposed for detecting the SARS-CoV-2 RNA with excellent sensitivity and selectivity. The Au NWs modified with bimetallic NPs can significantly increase the SERS/EC signals, and the presence of the SARS-CoV-2 RNA can produce significantly attenuated SERS/EC signals by shearing off the aptamer containing the signal molecules. In addition to its excellent specific recognition of the SARS-CoV-2 RNA and high sensitivity (detection limits of 1.02 pM by the SERS method and 0.21 pM by the EC method), this nanosensor has been successfully applied to samples with complex matrices and is expected to serve as a potential, sensitive, and reliable tool for virus screening and protein detection, especially for in vivo detection due to the small overall dimension of single Au NWs.

研究领域: 生物传感器 · 病毒检测 · 纳米技术
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ADVANCED FUNCTIONAL MATERIALS · 2022 ·Vol. 32 ·期 48 ·2209743 · IF 19.924

2D Zn-Porphyrin-Based Co(II)-MOF with 2-Methylimidazole Sitting Axially on the Paddle–Wheel Units: An Efficient Electrochemiluminescence Bioassay for SARS-CoV-2

Yi-Xuan Li,Jing Li,Dunru Zhu,Ju-Zheng Wang,Guo-Fang Shu,Junji Li,Sheng-Li Zhang,Xue-Ji Zhang,Serge Cosnier,Hai-Bo Zeng,Dan Shan

High electrocatalytic activity with tunable luminescence is crucial for the development of electrochemiluminescence (ECL) luminophores. In this study, a porphyrin-based heterobimetallic 2D metal organic framework (MOF), [(ZnTCPP)Co 2 (MeIm)] (1), is successfully self-assembled from the zinc(II) tetrakis(4-carboxyphenyl)porphine (ZnTCPP) linker and cobalt(II) ions in the presence of 2-methylimidazole (MeIm) by a facile one-pot reaction in methanol at room temperature. On the basis of the experimental results and the theoretical calculations, the MOF 1 contains paddle–wheel [Co 2 (-CO 2 ) 4 ] secondary building units (SBUs) axially coordinated by a MeIm ligand, which is very beneficial to the electron transfer between the Co(II) ions and oxygen. Combining the photosensitizers ZnTCPP and the electroactive [Co 2 (-CO 2 ) 4 ] SBUs, the 2D MOF 1 possesses an excellent ECL performance, and can be used as a novel ECL probe for rapid nonamplified detection of the RdRp gene of SARS-CoV-2 with an extremely low limit of detection (≈30 aM).

研究领域: 材料科学 · 分析化学 · 病毒学
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Polymers · 2022 ·Vol. 14 ·期 24 ·5466 · IF 4.967

Self-Assembled TLR7/8 Agonist-Mannose Conjugate as An Effective Vaccine Adjuvant for SARS-CoV-2 RBD Trimer

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

研究领域: 免疫学 · 病毒学 · 药学
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International Journal of Electrochemical Science · 2023 · IF 1.5

Electrochemical Biosensor for the Detection of SARS-CoV-2 Main Protease and Its Inhibitor Ebselen

Qiongyu Zhang,Gang Liu,Lingbin Ou

This work reported an electrochemical method for the detection of SARS-CoV-2 major protease (Mpro). Specifically, ferrocene (Fc)-labeled peptide substrates were immobilized on the gold nanoparticles (AuNPs)-modified electrode. Cleavage of the peptides by Mpro led to the release of Fc tags and the decrease of the electrochemical signals. The analytical performance of the biosensor for analysis of Mpro was investigated. Inhibiting the activity of Mpro prevented the cleavage of the peptide substrates. The method was successfully used to evaluate the inhibition efficiency of a well-known inhibitor.

研究领域: 生物化学 · 分析化学 · 病毒学
✓ 已验证 DOI PubMed 高影响因子
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MICROCHIMICA ACTA · 2022 ·Vol. 189 ·期 8 ·1-10 · IF 6.408

A smartphone-based platform for ratiometric visualization of SARS-CoV-2 via an oligonucleotide probe

Zhao Tingting,Yan Weizhen,Dong Fengqi,Hu Xinlong,Xu Yanli,Wang Zhenyu,Shen Yating,Wang Wanrong,Zhao Ye,Wei Wenmei

COVID-19 necessitates the development of reliable and convenient diagnostic tools. In this work, a facile 3D-printed smartphone platform was constructed that achieved reliable visual detection of SARS-CoV-2 by eliminating the effect of ambient light and fixing the camera position relative to the sample. The oligonucleotide probe is modified with orange-red-emitting TAMRA working as an internal standard and green-emitting FAM serving as a sensitive sensing agent. Under 365-nm UV excitation, the emission wavelengths of TAMRA and FAM are 580 nm and 518 nm, respectively. When the probes interact with the targets, the green fluorescence gradually restores while the orange-red fluorescence remains stable. Thus, a striking color transition from orange-red to green could be observed by the naked eye. The detection limit of SARS-CoV-2 nucleic acid is 0.23 nM, and the entire process of color change could be completed in 25 min. Furthermore, the RGB value analysis of the sample solution was conducted using a smartphone for reliable and reproducible discrimination of SARS-CoV-2. The proposed smartphone platform might establish a general method for visual detection of SARS-CoV-2 nucleic acid as well as other virus-related diseases. Graphical abstract

研究领域: 生物医学工程 · 分子生物学 · 诊断工具 · 病毒学
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CARBOHYDRATE POLYMERS · 2022 · IF 10.723

Inhibitory activity of a sulfated oligo-porphyran from Pyropia yezoensis against SARS-CoV-2

Lihua Geng,Quanbin Zhang,Qishan Suo,Jing Wang,Yingxia Wang,Cong Wang,Ning Wu

COVID-19 caused by SARS-CoV-2 has spread around the world at an unprecedented rate. A more homogeneous oligo-porphyran with mean molecular weight of 2.1 kD, named OP145, was separated from Pyropia yezoensis . NMR analysis showed OP145 was mainly composed of →3)-β- d -Gal-(1 → 4)-α- l -Gal (6S) repeating units with few replacement of 3,6-anhydride, and the molar ratio was 1:0.85:0.11. MALDI-TOF MS revealed OP145 contained mainly tetrasulfate-oligogalactan with Dp range from 4 to 10 and with no more than two 3,6-anhydro-α- l -Gal replacement. The inhibitory activity of OP145 against SARS-CoV-2 was investigated in vitro and in silico. OP145 could bind to Spike glycoprotein (S-protein) through SPR result, and pseudovirus tests confirmed that OP145 could inhibite the infection with an EC50 of 37.52 μg/mL. Molecular docking simulated the interaction between the main component of OP145 and S-protein. All the results indicated that OP145 had the potency to treat and prevent COVID-19.

CAS / MDL: 127-09-3
研究领域: 病毒学 · 药理学 · 分子生物学
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