Research
·
2022
·
IF 11.036
Song Liu,Guanghui Li,Lei Ding,Jin Ding,Qian Zhang,Dan Li,Xingguo Hou,Xiangxing Kong,Jing Zou,Shiming Zhang,Hongbin Han,Yakun Wan,Zhi Yang,Hua Zhu
Due to the rapid spread of coronavirus disease 2019 (COVID-19), there is an urgent requirement for the development of additional diagnostic tools for further analysis of the disease. The isolated nanobody Nb11-59 binds to the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor-binding domain (RBD) with high affinity to neutralize the virus and block the angiotensin-converting enzyme 2- (ACE2-) RBD interaction. Here, we introduce a novel nanobody-based radiotracer named 68Ga-Nb1159. The radiotracer retained high affinity for the RBD and showed reliable radiochemical characteristics both in vitro and in vivo. Preclinical positron emission tomography (PET) studies of 68Ga-Nb1159 in mice revealed its rapid clearance from circulation and robust uptake into the renal and urinary systems. Fortunately, 68Ga-Nb1159 could specifically reveal the distribution of the RBD in mice. This study also helped to evaluate the pharmacodynamic effects of the neutralizing nanobody. Moreover, 68Ga-Nb1159 may be a promising tool to explore the distribution of the RBD and improve the understanding of the virus. In particular, this study identified a novel molecular radioagent and established a reliable evaluation method for specifically investigating the RBD through noninvasive and visual PET technology.
研究领域:
医学影像学
·
病毒学
·
药理学
✓ 已验证
DOI
高影响因子
CHEMBIOCHEM
·
2024
·
IF 2.6
Yi Yang,Zhiyi Zhao,Xiaoying Li,Yian Chen,Lu Liu,Shao-Lin Zhang,Aimin Yang
Graphical Our study designs and synthesizes a series of peptides targeting the dimerization of SARS-CoV-2 M pro . Several peptides display potential inhibitory activities. Specifically, N8-A24 shows most efficient suppression of the M pro activity via inhibiting its dimerization. The severe acute respiratory syndrome virus 2 (SARS-CoV-2) seriously impacted public health. The evolutionarily conserved viral chymotrypsin-like main protease (M pro ) is an important target for anti-SARS-CoV-2 drug development. Previous studies have shown that the eight N-terminal amino acids (N8) of SARS-CoV M pro are essential for its dimerization, and are used to design inhibitors against SARS-CoV M pro dimerization. Here, we established a simple readout assay using SDS-PAGE and Coomassie blue staining to measure inhibitory activity of N8 peptide derived from SARS-CoV-2 M pro . To optimize its inhibitory effect, we then modified the side-chain length, charge, and hydrophilicity of the N8 peptide, and introduced a mutated M pro recognition sequence. As a result, we obtained a series of potent peptide inhibitors against SARS-CoV-2 M pro , with N8-A24 being the most efficient with an IC 50 value of 1.44 mM. We observed that N8-A24 reduced M pro dimerization with an IC 50 value of 0.86 mM. Molecular docking revealed that N8-A24 formed hydrogen bond interactions with critical dimeric interface residues, thus inhibiting its dimerization and activity. In conclusion, our study not only discovers a series of peptide inhibitors targeting the SARS-CoV-2 M pro dimerization, but also provides a promising strategy for the rational design of new inhibitors against COVID-19.
研究领域:
病毒学
·
药物开发
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
Foods
·
2022
·Vol. 11
·期 18
·2865
·
IF 5.561
Jinghe Sun,Shuang Song,Chunqing Ai,Beiwei Zhu,Jingfeng Yang
Sulfate polysaccharides, such as heparin sulfate, have been found to have inhibitory activity against SARS-CoV-2. An abalone polysaccharide, AGSP, was deeply sulfate modified using the chlorosulfonic acid/pyridine method, yielding S-AGSP. AGSP and S-AGSP inhibitions of SARS-CoV-2 infection of Vero E6 cells were tested in vitro. The interference of AGSP or S-AGSP on the binding interaction between the SARS-CoV-2 spike protein and angiotensin-converting enzyme was tested using a biolayer interferometry assay. Results showed that S-AGSP, above a concentration of 1.87 µg/mL, significantly inhibited SARS-CoV-2 infection of Vero E6 cells. Compared with AGSP, S-AGSP obviously weakened the affinity between the SARS-CoV-2 spike protein and ACE2. The polysaccharide’s sulfate content played a vital role in influencing the binding affinity of spike protein to ACE2. Therefore, S-AGSP has potential as a COVID-19 competitive inhibitor as well as a candidate to be repurposed as a prophylactic COVID-19 therapeutic.Keywords:sulfated;polysaccharide;SARS-CoV-2;biolayer interferometry;ACE2
研究领域:
病毒学
·
药理学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
Biosensors-Basel
·
2021
·Vol. 12
·期 1
·13
·
IF 5.519
Tao Peng,Xueshima Jiao,Zhanwei Liang,Hongwei Zhao,Yang Zhao,Jie Xie,You Jiang,Xiaoping Yu,Xiang Fang,Xinhua Dai
The coronavirus disease 2019 (COVID-19) pandemic caused by severe acute respiratory coronavirus 2 (SARS-CoV-2) is still raging all over the world. Hence, the rapid and sensitive screening of the suspected population is in high demand. The nucleocapsid protein (NP) of SARS-CoV-2 has been selected as an ideal marker for viral antigen detection. This study describes a lateral flow immunoassay (LFIA) based on colloidal gold nanoparticles for rapid NP antigen detection, in which sensitivity was improved through copper deposition-induced signal amplification. The detection sensitivity of the developed LFIA for NP antigen detection (using certified reference materials) under the optimized parameters was 0.01 μg/mL and was promoted by three orders of magnitude to 10 pg/mL after copper deposition signal amplification. The LFIA coupled with the copper enhancement technique has many merits such as low cost, high efficiency, and high sensitivity. It provides an effective approach to the rapid screening, diagnosis, and monitoring of the suspected population in the COVID-19 outbreak.Keywords:SARS-CoV-2;nucleocapsid protein;signal amplification;copper deposition
研究领域:
病毒学
·
医学诊断
·
纳米技术
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CELL
·
2021
·
IF 41.584
Liming Yan,Yunxiang Yang,Mingyu Li,Ying Zhang,Litao Zheng,Ji Ge,Yucen C. Huang,Zhenyu Liu,Tao Wang,Shan Gao,Ran Zhang,Yuanyun Y. Huang,Luke W. Guddat,Yan Gao,Zihe Rao,Zhiyong Lou
The capping of mRNA and the proofreading play essential roles in SARS-CoV-2 replication and transcription. Here, we present the cryo-EM structure of the SARS-CoV-2 replication-transcription complex (RTC) in a form identified as Cap(0)-RTC, which couples a co-transcriptional capping complex (CCC) composed of nsp12 NiRAN, nsp9, the bifunctional nsp14 possessing an N-terminal exoribonuclease (ExoN) and a C-terminal N7-methyltransferase (N7-MTase), and nsp10 as a cofactor of nsp14. Nsp9 and nsp12 NiRAN recruit nsp10/nsp14 into the Cap(0)-RTC, forming the N7-CCC to yield cap(0) ( 7Me GpppA) at 5′ end of pre-mRNA. A dimeric form of Cap(0)-RTC observed by cryo-EM suggests an in trans backtracking mechanism for nsp14 ExoN to facilitate proofreading of the RNA in concert with polymerase nsp12. These results not only provide a structural basis for understanding co-transcriptional modification of SARS-CoV-2 mRNA but also shed light on how replication fidelity in SARS-CoV-2 is maintained.
研究领域:
病毒学
·
结构生物学
·
分子生物学
✓ 已验证
DOI
PubMed
高影响因子
FEBS Open Bio
·
2022
·Vol. 12
·期 10
·1886-1895
·
IF 2.792
Yufang Zhang,Wenkang Li,Yiming Hu,Tianze Ding,Muhammad Mubashar Zafar,Xue Jia,Liya Zhang,Maozhi Ren,Fuguang Li,Wenjing Wang
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been spreading globally for over 2 years, causing serious contagious disease and incalculable damage. The introduction of vaccines has slowed the spread of SARS-CoV-2 to some extent, but there remains a need for specific and effective treatment. The high chemical diversity and safety profiles of natural products make them a potential source of effective anti-SARS-CoV-2 drugs. Cotton plant is one of the most important economic and medical crops and is the source of a large number of antiviral phytochemicals. In this work, we used SARS-CoV-2 main protein (M pro ) as the target to identify potential anti-SARS-CoV-2 natural products in cotton. An in vitro assay showed that of all cotton tissues examined, cotton flower extracts (CFs) exhibited optimal inhibitory effects against M pro . We proceeded to use the CF metabolite database to screen natural M pro inhibitors by combining virtual screening and biochemical assays. We identified that several CF natural products, including astragalin, myricitrin, and astilbin, significantly inhibited M pro with half-maximal inhibitory concentrations (IC50s) of 0.13, 10.73, and 7.92 μ m , respectively. These findings may serve as a basis for further studies into the suitability of cotton as a source of potential therapeutics for SARS-CoV-2.
研究领域:
病毒学
·
天然药物化学
·
分子药理学
·
植物生物技术
·
抗病毒治疗
·
药物发现
✓ 已验证
DOI
高影响因子
ACS Applied Materials & Interfaces
·
2023
·
IF 9.5
Zhongjie Fei,Ping Liu,Chu Cheng,Rongbin Wei,Pengfeng Xiao,Youfa Zhang
Although numerous approaches were proposed for the nucleic acid (NA)-based SARS-CoV-2 detection, the nonideal NA desorption efficiency of conventional magnetic beads (MBs) limits their widespread application. In this study, we developed solvent-responsive MBs (called responsive MBs), which, in the presence of buffers, modulated the absorption and desorption capacities of NA by flipping the surface −COO–. Relative to other commercial MBs, responsive MBs exhibited similar absorption profiles and markedly enhanced desorption profiles. When applied for NA detection of complex samples, responsive MBs exhibited better performance of RNA detection than DNA, with obvious advantages in sensitivity. Specifically, the RNA and DNA desorption rates of commercial MBs were ∼85 and 82.5%, while those of responsive MBs were nearly 94 and 93.5%, respectively. Furthermore, responsive MBs exhibited remarkable extraction ability in a wide range of tissues and better performance of RNA extraction than DNA. When applied for SARS-CoV-2 detection, the responsive MBs along with the simulated digital RT-LAMP (a previously established apparatus) further improved detection efficiency, yielding a precise quantitative detection as low as 25 copies and an ultimate sensibility detection of 5 copies/mL. It was also successfully employed in numerous NA-based technologies such as polymerase chain reaction (PCR), sequencing, and so on.
研究领域:
生物技术
·
分子生物学
·
病毒学
·
分析化学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2021
·
IF 10.618
Ying Peng,Yanhong Pan,Zhaowei Sun,Jinlong Li,Yongxiang Yi,Jie Yang,Genxi Li
The pandemic of coronavirus disease 2019 (COVID-19) caused by the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) is continuously worsening globally, herein we have proposed an electrochemical biosensor for the sensitive monitoring of SARS-CoV-2 RNA. The presence of target RNA firstly triggers the catalytic hairpin assembly circuit and then initiates terminal deoxynucleotidyl transferase-mediated DNA polymerization. Consequently, a large number of long single-stranded DNA products can be produced, and these negatively charged DNA products will bind a massive of positively charged electroactive molecular of Ru(NH 3 ) 6 3+ due to the electrostatic adsorption. Therefore, significantly amplified electrochemical signals can be generated for sensitive analysis of SARS-CoV-2 RNA in the range of 0.1–1000 pM with the detection limit as low as 26 fM. Besides the excellent distinguishing ability for SARS-CoV-2 RNA against single-base mismatched RNA, the proposed biosensor can also be successfully applied to complex matrices, as well as clinical patient samples with high stability, which shows great prospects of clinical application.
研究领域:
分子生物学
·
分析化学
·
医学诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PROTEIN EXPRESSION AND PURIFICATION
·
2022
·
IF 2.025
Yi Rong,Chaofeng Zhang,Wen-Chao Gao,Cheng Zhao
The main protease (M pro ) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) plays a vital role in viral replication. To study the function of M pro and screen inhibitors targeting M pro , it is necessary to prepare high-purity and high-activity M pro . In this study, four types of SARS-CoV-2 M pro s containing different termini were prepared, and their activities were determined successfully. The results showed that the activity of wild-type (WT) M pro was the highest, and the additional residues at the N-terminus but not at the C-terminus had a major effect on the enzyme activity. To explain this, the alignment of structures of different forms of M pro was determined, and the additional residues at the N-terminus were found to interfere with the formation of the substrate binding pocket. This study confirms the importance of the natural N-terminus to the activity of M pro and suggests that WT-GPH 6 (M pro with eight additional residues at the C-terminus) can be used as a substitute for authentic M pro to screen inhibitors. In short, this study provides a reference for the expression and purification of new coronaviruses confronted in the future.
研究领域:
病毒学
·
生物化学
·
分子生物学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2024
·
IF 5.7
Yuping Zeng,Xiangyu Gan,Zhourui Xu,Xiaoxiang Hu,Chenxuan Hu,Hanbin Ma,Hangjia Tu,Bao Chai,Chengbin Yang,Siyi Hu,Yujuan Chai
Background Sensitive and rapid antigen detection is critical for the diagnosis and treatment of infectious diseases, but conventional ELISAs including chemiluminescence-based assays are limited in sensitivity and require many operation steps. Fluorescence immunoassays are fast and convenient but often show limited sensitivity and dynamic range. Results To address the need, an aggregation-induced emission fluorgens (AIEgens) enhanced i mmunofluorescent assay with beads-based quantification on the digital microfluidic (DMF) platform was developed. Portable DMF devices and chips with small electrodes were fabricated, capable of manipulating droplets within 100 nL and boosting the reaction efficiency. AIEgen nanoparticles (NPs) with high fluorescence and photostability were synthesized to enhance the test sensitivity and detection range. The integration of AIEgen probes, transparent DMF chip design, and the large magnetic beads (10 μm) as capture agents enabled rapid and direct image-taking and signal calculation of the test result. The performance of this platform was demonstrated by point-of-care quantification of SARS-CoV-2 nucleocapsid (N) protein. Within 25 min, a limit of detection of 5.08 pg mL −1 and a limit of quantification of 8.91 pg mL −1 can be achieved using <1 μL sample. The system showed high reproducibility across the wide dynamic range (10–10 5 pg mL −1 ), with the coefficient of variance ranging from 2.6% to 9.8%. Significance This rapid, sensitive AIEgens-enhanced immunofluorescent assay on the DMF platform showed simplified reaction steps and improved performance, providing insight into the small-volume point-of-care testing of different biomarkers in research and clinical applications.
研究领域:
生物医学工程
·
分析化学
·
分子诊断
✓ 已验证
产品关联
DOI
PubMed
高影响因子
CELL RESEARCH
·
2021
·Vol. 31
·期 10
·1047-1060
·
IF 25.617
Hu Wei,Zhang Yong,Fei Panyu,Zhang Tongtong,Yao Danmei,Gao Yufei,Liu Jia,Chen Hui,Lu Qiao,Mudianto Tenny,Zhang Xinrui,Xiao Chuxuan,Ye Yang,Sun Qiming,Zhang Jing,Xie Qi,Wang Pei-Hui,Wang Jun,Li Zhenhai,Lou Jizhong,Chen Wei
The outbreak of SARS-CoV-2 (SARS2) has caused a global COVID-19 pandemic. The spike protein of SARS2 (SARS2-S) recognizes host receptors, including ACE2, to initiate viral entry in a complex biomechanical environment. Here, we reveal that tensile force, generated by bending of the host cell membrane, strengthens spike recognition of ACE2 and accelerates the detachment of spike’s S1 subunit from the S2 subunit to rapidly prime the viral fusion machinery. Mechanistically, such mechano-activation is fulfilled by force-induced opening and rotation of spike’s receptor-binding domain to prolong the bond lifetime of spike/ACE2 binding, up to 4 times longer than that of SARS-S binding with ACE2 under 10 pN force application, and subsequently by force-accelerated S1/S2 detachment which is up to ~10 3 times faster than that in the no-force condition. Interestingly, the SARS2-S D614G mutant, a more infectious variant, shows 3-time stronger force-dependent ACE2 binding and 35-time faster force-induced S1/S2 detachment. We also reveal that an anti-S1/S2 non-RBD-blocking antibody that was derived from convalescent COVID-19 patients with potent neutralizing capability can reduce S1/S2 detachment by 3 × 10 6 times under force. Our study sheds light on the mechano-chemistry of spike activation and on developing a non-RBD-blocking but S1/S2-locking therapeutic strategy to prevent SARS2 invasion.
研究领域:
病毒学
·
分子生物学
·
生物物理学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
ANALYST
·
2023
·Vol. 148
·期 15
·3603-3609
·
IF 4.2
Ju-Zheng Wang,Yi-Xuan Li,Qiaoting Yang,Junji Li,Jérome Chauvin,Xue-Ji Zhang,Serge Cosnier,Robert S. Marks,Dan Shan
The emergence of novel pathogens, as well as their frequent variants, raises the significance of developing superior and versatile sensing materials and techniques. Herein, a post-modified zeolitic imidazolate framework (pm-ZIF) was synthesized by using ZIF-67 as a parent MOF, and zinc(II) meso-tetra (4-carboxyphenyl) porphine (ZnTCPP) as a successive exchange ligand. Due to the preservation of the tetrahedral Co–N4 units from the ZIF precursor and the introduced porphyrin luminophores, this hybrid material pm-ZIF/P(Zn) enables the linear electrochemiluminescence (ECL) signal conversion of the target DNA concentration. An efficient biosensor that can be used to quantitatively detect SARS-CoV-2 was therefore constructed. The linear range of the sensor was 10−12–10−8 M, with a limit of detection (LOD) reaching 158 pM. Compared with the traditional amplification-based methods, the duration time of our method is significantly shortened and the quantitation of the SARS-Cov-2 RdRp gene can be completed within twenty minutes at room temperature.
研究领域:
材料科学
·
生物医学工程
·
病毒学
✓ 已验证
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）