Micromachines
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2018
·Vol. 9
·期 6
·274
·
IF 2.222
Zihao Li,Chenggang Zhu,Zhihe Guo,Bowen Wang,Xiang Wu,Yiyan Fei
The detection of small molecules has increasingly attracted the attention of researchers because of its important physiological function. In this manuscript, we propose a novel optical sensor which uses an optofluidic microbubble resonator (OFMBR) for the highly sensitive detection of small molecules. This paper demonstrates the binding of the small molecule biotin to surface-immobilized streptavidin with a detection limit reduced to 0.41 pM. Furthermore, binding specificity of four additional small molecules to surface-immobilized streptavidin is shown. A label-free OFMBR-based optical sensor has great potential in small molecule detection and drug screening because of its high sensitivity, low detection limit, and minimal sample consumption.Keywords:label-free sensor;optofluidic microbubble resonator;detection of small molecules
研究领域:
生物医学工程
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光学传感
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分子生物学
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DOI
PubMed
高影响因子
BioScience Trends
·
2014
·Vol. 7
·期 3
·144-151
·
IF 1.213
Xiaoying Zhou,Yazhou Cui,Jing Luan,Xiaoyan Zhou,Genglin Zhang,Xiumei Zhang,Jinxiang Han
Matrix vesicles (MVs) involved in the initiation of mineralization by deposition of hydroxyapatite (HA) in their lumen are released by the budding of mineralization-competent cells during skeletogenesis and bone development. To identify additional mineralization-related proteins, MVs were isolated from non-stimulated and stimulated Saos-2 cells in culture via an ExoquickTM approach and the corresponding proteomes were identified and quantified with label-free quantitative proteome technology. The isolated MVs were confirmed by electron microscopy, alkaline phosphatase activity (ALP), biomarkers, and mineral formation analyses. Label-free quantitative proteome analysis revealed that 19 calcium binding proteins (CaBPs), including Grp94, calnexin, calreticulin, calmodulin, and S100A4/A10, were up-regulated in MVs of Saos-2 cells upon stimulation of mineralization. This result provides new clues to study the mechanism of the initiation of MV-mediated mineralization.
研究领域:
蛋白质组学
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骨生物学
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细胞生物化学
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DOI
高影响因子
RSC Advances
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2022
·Vol. 12
·期 46
·30024-30029
·
IF 4.036
Baozhu Zhang,Ziyao Yang,Yuxia Li,Ling Ma,Fenfang Li,Xiuqing Lv,Guangming Wen
A label-free aptasensor has been fabricated in order to detect adenosine triphosphate (ATP) using turn-on fluorescence DNA-Ag NCs. The fluorescence of the DNA-Ag NCs could increase remarkably with the addition of ATP mainly because ATP specifically interacts with its aptamer to change the microenvironment of the darkish DNA-Ag NCs located at one terminus or two termini due to the conformational alteration of the aptamer structure. The proposed sensor can detect ATP in a linear range of 6–27 mM with a good detection limit of 5.0 μM. Additionally, the proposed method succeeded in detecting ATP in fetal bovine serum.
研究领域:
分析化学
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生物传感器
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纳米技术
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DOI
PubMed
高影响因子
STAR Protocols
·
2025
·
IF 1.3
Yu Ran,Qiqing Yang,Lianqi Zhou,Heyu Li,Bing Yang,Long Zhang
Summary L-lactate has been recognized as an essential molecule for signaling and metabolic balance. Lactylation, a post-translational modification (PTM) derived from L-lactate, is commonly observed on various proteins and plays essential roles in cellular processes. Here, we present a protocol to globally profile the lactylation proteome and perform label-free quantification. We provide steps for cell preparation, protein extraction, digestion, peptide desalting, and lactylpeptide enrichment. Additionally, we outline the parameters for liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. For complete details on the use and execution of this protocol, please refer to Li et al. 1
研究领域:
生物化学
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蛋白质组学
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分子生物学
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DOI
PubMed
高影响因子
ANALYTICAL CHEMISTRY
·
2024
·
IF 6.7
Shen Jiang,Qiuyun Li,Guangrun Wu,Xuming Mu,Xiaotong Wang,Yunpeng Wang,Yanli Wu,Jing Wu,Yang Li
In the landscape of biomolecular detection, surface-enhanced Raman spectroscopy (SERS) confronts notable obstacles, particularly in the label-free detection of biomolecules, with glucose and other sugars presenting a quintessential challenge. This study heralds the development of a pioneering SERS substrate, ingeniously engineered through the self-assembly of nanoparticles of diverse sizes (Ag1@Ag2NPs). This configuration strategically induces ‘hot spots’ within the interstices of nanoparticles, markedly amplifying the detection signal. Rigorous experimental investigations affirm the platform’s rapidity, precision, and reproducibility, and the detection limit of this detection method is calculated to be 6.62 pM. Crucially, this methodology facilitates nondestructive glucose detection in simulated samples, including phosphate-buffered saline and urine. Integrating machine learning algorithms with simulated serum samples, the approach adeptly discriminates between hypoglycemic, normoglycemic, and hyperglycemic states. Moreover, the platform’s versatility extends to the detection and differentiation of monosaccharides, disaccharides, and methylated glycosides, underscoring its universality and specificity. Comparative Raman spectroscopic analysis of various carbohydrate structures elucidates the unique SERS characteristics pertinent to these molecules. This research signifies a major advance in nonchemical, label-free glucose determination with enhanced sensitivity via SERS, laying a new foundation for its application in precision medicine and advancing structural analysis in the sugar domain.
研究领域:
生物医学工程
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分析化学
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纳米技术
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2024
·
IF 10.7
Jing Ye,Qi Liang,Qianglong Tan,Mengyao Chai,Wendai Cheng,Minzhi Fan,Yunshan Zhang,Jie Zhan,Yaxin Wang,Jiahong Wen,Yongjun Zhang,Xiaoyu Zhao,Diming Zhang
Compared to conventional nucleic acid detection methods, label-free single nucleotide polymorphism (SNP) detection presents challenging due to the necessity of discerning single base mismatches, especially in the field of enzyme-free detection. In this study, we introduce a novel bulged-type DNA duplex probe designed to significantly amplify single-base differences. This probe is integrated with programmable DNA-based nanostructures to develop a sensitive, label-free biosensor for nonenzymatic SNP detection. The duplex probe with one bulge could selectively identify wild-typed DNA (WT) and mutant-type DNA (MT) based on a competitive strand displacement reaction mechanism. The hyperbranched HCR (HHCR) by incorporating of hairpin DNA into the DNA tetrahedron and surface-tethering on the portable screen printing electrode (SPCE) significantly favor the formation of negatively charged DNA nanostructure. We harnessed strong repulsion of DNA nanostructure towards the electroactive [Fe(CN)₆]³⁻/⁴⁻ in combination with electrochemical technique to create a label-free biosensor. This simple, enzyme-free and label-free biosensor could detect MT with a detection limit of 56 aM, even in multiple sequence backgrounds. The study served as the proof-of-concept for the integration of enzyme-free competitive mechanism and label-free strategy, which can be extended as a powerful tool to various fields.
研究领域:
生物化学
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分子生物学
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分析化学
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DOI
PubMed
高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2018
·
IF 5.667
Bingzhi Li,Yue Chen,Jing Wang,Qiaoyun Lu,Wanying Zhu,Lei Xu,Xin Shen,Jieping Luo,Chunhong Zhu,Xiaoxu Li,Junli Hong,Xuemin Zhou
Transcription factors (TFs) regulate information flow from gene to protein, and they are recognized as key indicators to reflect cellular processes. Facile monitoring of TFs may aid diagnostics and treatment, but we still lack of techniques for highly efficient detection of them. In this research, a hybrid material consisting of DNA-silver nanoclusters (DNA-AgNCs) and polypyrrole nanoparticles (PPyNPs) was built for TFs detection. The designed DNA-AgNCs have a hairpin-shaped nucleic acid architecture with a double-stranded stem for recognizing TFs and a single-stranded loop for interacting with PPyNPs. In the absence of TFs, DNA-AgNCs are absorbed to PPyNPs, resulting in fluorescent quenching. While in the presence of TFs, the binding between TFs and the DNA-AgNCs caused desorption of DNA-AgNCs via steric hindrance mechanism. Accordingly, the increase of fluorescence derived from desorption is used for quantifications. Derived from low non-specific protein absorption features of PPyNPs, detection limit of 70 pM for NF-κB p50 and 110 pM for p53 were obtained. Then, by absorbing two kinds of DNA-AgNCs to PPyNPs, label-free multiplexed detection of TFs was first realized. Additionally, we suggested that this platform can be developed for drug screening by evaluating inhibitory effect of a pair of optical isomers towards TFs.
研究领域:
纳米技术
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生物医学工程
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分析化学
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分子生物学
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DOI
高影响因子
ANALYTICAL CHEMISTRY
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2020
·
IF 6.785
Guizhen Liu,Tao Fu,Ying Han,Shichen Hu,Xuepei Zhang,Mengmeng Zheng,Piliang Hao,Lifeng Pan,Jingwu Kang
We describe an affinity purification–mass spectrometry (AP–MS) method for probing the interactome of a special targeting protein. The AP was implemented with monolithic micro immobilized metal ion affinity chromatography columns (m-IMAC) which were prepared by photoinitiated polymerization in the tip of a pipet (spin-tip columns). The recombinant His6-tagged protein (bait protein) was reversibly immobilized on the affinity column through the chelating group nitrilotriacetic acid (NTA)–Ni2+. The bait protein and its interacting partners can be easily eluted from the affinity matrix. The pulled-down cellular proteins were then analyzed with label-free quantitative proteomics. We used this method for probing the interactome concerning the GOLD (Golgi dynamics) domain of the autophagy-associated adaptor protein FYCO1. Totally, 96 proteins including seven literature-reported FYCO1-associating proteins were identified. Among them CCZ1 and MON1A were further biochemically validated, and the direct interaction between the FYCO1 GOLD domain with CCZ1 was confirmed by co-immunoprecipitation experiments.
研究领域:
蛋白质组学
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生物化学
·
细胞生物学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
·
2024
·
IF 5.7
Chiyu Ma,Yong Qiu,Tao Liang,Nan Jiang,Liubing Kong,Jianguo Wu,Xianyou Sun,Guangqing Ren,Xinwei Wei,Ping Wang,Hao Wan
Background Ionic calcium (Ca 2+ ) plays a crucial role in maintaining normal physiological and biochemical functions within the human body. Detecting the concentration of Ca 2+ is of utmost significance for various purposes, including disease screening, cellular metabolism research, and evaluating drug effectiveness . However, current detection approaches such as fluorescence and colorimetry face limitations due to complex labeling techniques and the inability to track changes in Ca 2+ concentration. In recent years, extensive research has been conducted in this field to explore label-free and efficient approaches. Results In this study, a novel light-addressed potentiometric sensor (LAPS) using silicon-on-sapphire technology , has been successfully developed for Ca 2+ sensing. The Ca 2+ -sensitive LAPS achieved a wide-range detection of Ca 2+ , ranging from 10 −2 M to 10 −7 M, with an impressive detection limit of 100 nM. These advancements are attributed to the ultra-thin silicon layer, silicon dioxide layer, and solid-state silicon rubber sensitive membrane around 6 μm. Furthermore, the sensor demonstrated the ability to dynamically monitor fluctuations in Ca 2+ concentration ranging from 10 −9 M to 10 −2 M within a solution. Its remarkable selectivity , specificity, and long-term stability have facilitated its successful application in the detection of Ca 2+ in human serum and urine. Significance and novelty This work presents a Ca 2+ -sensitive sensor that combines a low detection limit and a wide detection range. The development represents the emergence of a label-free and rapid Ca 2+ detection tool with immense prospects in home-based health monitoring, community disease screening, as well as cellular metabolism, and drug screening evaluations.
研究领域:
生物医学工程
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分析化学
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传感器与执行器
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医学诊断
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DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
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2022
·
IF 5.304
Li Fu,Qingmei Chen,Suqi Deng,Li Jia
An economical and label-free fluorescent sensor was developed for the selective detection of Escherichia coli ( E. coli ) based on the competitive reduction of Cu 2+ in CuFe 2 O 4 magnetic particles (MPs) by E. coli and o -phenylenediamine (OPD). Cu 2+ in CuFe 2 O 4 MPs can oxidize OPD to generate 2,3-diaminophenazine (OPDox) with fluorescent property. The presence of E. coli can weaken the oxidative ability of CuFe 2 O 4 MPs toward OPD owing to the copper reductase activity of NADH dehydrogenase-2 (NDH-2) in E. coli . NDH-2 can reduce Cu 2+ to Cu + , thereby inhibiting the OPD oxidation and resulting in the decrease of the fluorescence of the system. Under the optimum conditions, the fluorescent sensor exhibited a wide linear response to E. coli in the concentration range of 10 3 – 10 6 cfu mL −1 , and the detection limit was calculated to be 5.8 × 10 2 cfu mL −1 . Furthermore, the feasibility of the sensor was verified by successful detection of E. coli in popsicle and swimming pool water samples. More importantly, the sensor utilized the magnetic property of CuFe 2 O 4 MPs to realize the phase separation, thus averting the interference of other impurities in the sample matrix.
研究领域:
分析化学
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生物技术
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环境科学
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微生物学
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DOI
高影响因子
CELLULAR SIGNALLING
·
2023
·
IF 4.8
Sijia Han,Xueying Jin,Tianyu Hu,Feng Chi
Non-SMC condensin II complex subunit G2 (NCAPG2) is one of the three non-SMC subunits in condensin II, which plays a vital role in regulating chromosome condensation and segregation. Although the tumor-promoting role of NCAPG2 has been reported in several solid malignancies, its function in breast invasive carcinoma (BRCA) remains unknown. Data both from GEPIA and GSE36295 indicated that NCAPG2 mRNA expression was abnormally upregulated in cancer tissues, which was further verified in 40 paired BRCA and para-carcinoma samples. Kaplan-Meier Plotter further illustrated that BRCA patients with higher NCAPG2 expression have a poorer prognosis. Functional experiments carried out in two BRCA cell lines (MCF-7 and T-47D) showed that NCAPG2-silenced BRCA cells acquired less aggressive behavior — weakened growth and metastasis both in vitro and in vivo. Label-free proteomics quantified the protein expression patterns in MCF-7 cells, and the results revealed 684 differentially expressed proteins (|log 2 FC| > 1 and P < 0.05) downstream to NCAPG2. Interestingly, poly(C)-binding protein 2 (PCBP2), an RNA binding protein previously known to increase RNA stability of its target genes, was found to directly bind to and protect NCAPG2 mRNA from degradation—PCBP2 knockdown accelerated the degradation half-life time of NCAPG2 mRNA from approximately 8 h to 5 h. Taken together, our study indicates that NCAPG2 acts as a novel contributor to BRCA growth and metastasis under the regulation of PCBP2, providing insights into BRCA treatment.
研究领域:
癌症生物学
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分子生物学
·
基因组学
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蛋白质组学
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细胞生物学
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产品关联
DOI
PubMed
高影响因子
COLLOIDS AND SURFACES B-BIOINTERFACES
·
2016
·
IF 3.902
Yu Li,Qi Wang,Yuting Zhang,Dongmei Deng,Haibo He,Liqiang Luo,Zhenxin Wang
A novel label-free electrochemical impedance aptasensor based on a gold nanoparticles/double-stranded DNA-graphene (AuNPs/dsDNA-GO) nanocomposite modified glassy carbon electrode was presented for quantitative determination of thrombin. GO was covalently functionalized with dsDNA via a facile amidation process, and then AuNPs were electrodeposited onto the surface of dsDNA-GO. The morphology, conductivity and interaction of the as-prepared nanocomposites were characterized by scanning electron microscopy, cyclic voltammetry, electrochemical impedance spectroscopy (EIS), Raman and Fourier transform infrared spectroscopy. The thrombin-binding aptamer (TBA) was conjugated to AuNPs via gold-thiol chemistry to construct electrochemical aptasensing platform, and the specific recognition between TBA and thrombin was monitored by EIS. Under optimum conditions, thrombin could be quantified in a wide range of 0.1–100 nM ( R 2 = 0.9960) with low detection limit of 0.06 nM (S/N = 3).
研究领域:
分析化学
·
纳米技术
·
生物传感器
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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）