MICROCHEMICAL JOURNAL
·
2024
·
IF 4.9
Yuanxun Gong,Jiayi Zhang,Huiying Liang,Yanling Wang,Zichun Song,Jihua Wei,Qianli Tang,Kai Zhang,Xianjiu Liao
Matrix metalloproteinases (MMPs) are critical for the degradation of extracellular matrix components, which is essential for processes such as tissue remodeling, wound healing, and angiogenesis. MMP-2, in particular, degrades type IV collagen and is associated with diseases such as cancer and cardiovascular conditions, making it a significant biomarker. This study presents a novel electrochemical biosensor for the detection of MMP-2. The biosensor combines the specificity of a programmable RNA detection system with the amplification capabilities of T7 RNA polymerase and CRISPR/Cas12a. In the absence of MMP-2, Peptide Nucleic Acid (PNA) adsorbs on the DNA template, preventing RNA amplification and resulting in a low electrochemical signal. When MMP-2 is present, it cleaves a specific peptide, releasing the DNA template and enabling RNA synthesis. This activation leads to the cleavage of single-stranded DNA probes, increasing the electrochemical signal. The biosensor demonstrates high sensitivity with a detection limit of 0.496 fM for MMP-2 and excellent specificity, effectively distinguishing MMP-2 from other proteins. It successfully detected MMP-2 in biological samples, such as liver cancer cell supernatants, indicating its potential for clinical diagnostics. This innovative biosensor offers a powerful tool for the early detection and monitoring of diseases associated with MMP-2 activity.
研究领域:
生物医学工程
·
分子诊断
·
纳米技术
✓ 已验证
产品关联
DOI
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Xiao Fang Zhang,Na Li,Yu Ling,Nian Bing Li,Hong Qun Luo
Since uracil-DNA glycosylase (UDG) is closely related to some human diseases, monitoring and detection of UDG activity have great significance in its clinic diagnosis and functional study. Here we demonstrate a sensitive cascade multi-amplification fluorescence strategy for the active UDG assay with the help of exonuclease III (Exo III). Under the cleavage reaction of UDG, the double-stranded DNA containing peculiar uracil bases separated into two dissociative single-stranded DNA, which separately hybridized with two cyclic hairpin probes to launch the Exo III-assisted dual cycles. The hairpin DNA modified with fluorophore and quenching group was employed as the signal probe. After the dual cycles, countless same short DNA fragments were generated to hybridize with signal probes, initiating a new Exo III-assisted cyclic amplification and releasing numerous single-stranded DNA which carried only fluorophores. Thus, the fluorescence intensity of the detection system was enhanced. This study obtained the detection limit as low as 2.4 × 10 −4 U/mL for detecting the UDG activity. And it performed satisfactory selectivity and well practical applicability by analysis of the HeLa cell lysate, providing a potential method for clinic diagnosis and functional study of UDG activity.
研究领域:
生物化学
·
分子生物学
·
临床诊断
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF DRUG DELIVERY SCIENCE AND TECHNOLOGY
·
2022
·
IF 5.062
Ying Zhang,Hongjun Yan,Rixiang Su,Peiyuan Li,Fangzhou Wen,Yingbin Lv,Jinyun Cai,Wei Su
Photodynamic therapy is a promising alternative to antibiotics in the treatment of bacterial infection, which has the advantages of less side effects, no drug resistance and accurate treatment. However, most photosensitizers (PSs) have some disadvantages, such as poor water solubility, serious self-quenching, poor stability and so on. Therefore, solving the limitations of traditional photosensitizers has become a hot issue in photodynamic therapy. On this basis, the lysozyme (Lys)-gold nanocluster (AuNCs)-curcumin (Cur) conjugate was used as a new type of water-soluble photosensitizer and combined with the bacterial surface through electrostatic interaction to form a bacterial surface enriched in PSs. At the same time, real-time bacterial imaging can be achieved by using the fluorescence properties of Lys-AuNCs-Cur. Under 405 nm light and dark conditions, the inactivation effect of Lys-AuNCs-Cur on Staphylococcus aureus and Escherichia coli was higher than that of free Cur. Moreover, Lys-AuNCs-Cur can also produce enough ROS to kill methicillin-resistant Staphylococcus aureus . The excellent antibacterial properties of nanoparticles may be attributed to the synergism of antibacterial protein lysozyme and AuNCs-Cur nanocomplexes and the increase of 1 O 2 production. The results of scanning electron microscope and fluorescence microscopic imaging showed that the cell membrane of E. coli and S. aureus was seriously damaged, and facilitated penetration of Lys-AuNCs-Cur nanocomplexes into the bacteria through the damaged cell membrane, and improve the photoinactivation efficiency of bacteria. More importantly, nanoparticles exhibited almost negligible dark cytotoxicity. The coupling of Cur and gold nanoclusters can significantly improve the stability of free curcumin molecules and the ability to generate ROS. Therefore, this work is of great significance for the development of new antimicrobial agents against bacterial infection.
研究领域:
纳米技术
·
生物医学工程
·
药学
·
微生物学
·
治疗学
✓ 已验证
产品关联
DOI
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2013
·
IF 5.437
Sai Bi,Yangyang Cui,Ying Dong,Ningbo Zhang
A biosensing system is established for the multi-amplified detection of DNA or specific substrates of aptamers under isothermal conditions , which combines nicked rolling circle amplification (N-RCA) and beacon assisted amplification (BAA) with sensitive colorimetric technique by using DNAzymes as reporter units. According to the configuration, the analysis of DNA is accomplished by recognizing the target to capture nucleic acid-functionalized magnetic particles, followed by the self-assembly of the other two nucleic acids into multicomponent DNA supramolecular structure on magnetic particles. After magnetic separation, the circularization with ligase and the fragmentation with polymerase activate N-RCA and BAA in the presence of polymerase, dNTPs , and the nicking endonuclease , successively producing horseradish peroxidase (HRP)-mimicking DNAzymes that act as colorimetric reporter to catalyze the oxidation of ABTS 2− by H 2 O 2 in the presence of hemin . Under the optimized conditions, we obtain a wide dynamic range for DNA analysis over 6 orders of magnitude from 1.0×10 −14 to 1.0×10 −9 M with a low limit of detection of 6.8×10 −15 M. In the absence of a target, neither self-assembly of nucleic acids nor amplification process can be initiated, indicating an excellent selectivity of the proposed strategy. Similarly, an analogous system is activated by cancer cells or lysozyme through cooperative self-assembly of nucleic acids on magnetic particles in the presence of respective substrates of aptamers to synthesize HRP-mimicking DNAzymes that give the readout signal for the recognition events, achieving LODs of 81 Ramos cells and 7.2×10 −15 M lysozyme, respectively.
研究领域:
生物传感
·
纳米技术
·
分子生物学
·
分析化学
✓ 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）