Analyst最新文献_第3页

Tyramide Signal Amplification for Highly Sensitive Multiplex Immunoassay based on Encoded Hydrogel Microparticles

IF 4.2 3区化学

Analyst Pub Date : 2025-03-26 DOI: 10.1039/d5an00078e

Jun Hee Choi, Young Hee Kim, Jiwoo Kim, Yong Jun Lim, Min Jung Kim, Ki Wan Bong

{"title":"Tyramide Signal Amplification for Highly Sensitive Multiplex Immunoassay based on Encoded Hydrogel Microparticles","authors":"Jun Hee Choi, Young Hee Kim, Jiwoo Kim, Yong Jun Lim, Min Jung Kim, Ki Wan Bong","doi":"10.1039/d5an00078e","DOIUrl":"https://doi.org/10.1039/d5an00078e","url":null,"abstract":"Proteins play a crucial role as mediators of immune regulation, homeostasis, and metabolism, making their quantification essential for understanding the disease mechanisms in biomedical research and clinical diagnostics. However, conventional methods have difficulty when detecting proteins in clinical samples in terms of sensitivity, dynamic range, and multiplex capacity. In this study, we develop a highly sensitive multiplex immunoassay based on encoded hydrogel microparticles (MP) utilizing tyramide signal amplification (TSA). The combination of large multiplexing capacity of encoded hydrogel microparticles and the signal amplification of TSA enables highly sensitive multiplex immunoassay. By employing the TSA, we are able to achieve bigger detection signals with higher specificity. We effectively decrease the non-specific bindings in the hydrogel network by blocking the unreacted acrylate double bonds remaining after the capture antibody conjugation step and acquire 3-fold increased signal-to-noise ratio. Also, we optimize three parameters mainly affecting the assay sensitivity: the detection antibody concentration, the biotinyl tyramide concentration and the TSA reaction time. This approach leads to a significant improvement in the assay sensitivity, achieving a limit of detection as low as 58 fg/mL. Compared to the previous method, the assay sensitivity is enhanced 10-fold. In addition, the multiplex capability of the assay is validated by detecting cytokines IL-4, IL-5, IL-6, IL-9, and IL-17, with no observed cross-reactivity. Finally, with enhanced sensitivity, we demonstrate the clinical applicability of our platform by successfully multiplexing these cytokines at concentrations down to several hundreds of fg/mL within the human serum, which could not be detected using previous methods.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"108 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702960","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Paper and Cloth-Based Microfluidic Chip for Rapid Cysteine Detection in Deep-Sea Cold Seep

IF 4.2 3区化学

Analyst Pub Date : 2025-03-26 DOI: 10.1039/d5an00109a

Die Hu, Jiawen Xiang, Jingying Guo, Chao Wang, Ji Qi, Bowei Li, Xiaoyan Wang, Xin Zhang, Lingxin Chen, Xuming Zhuang

引用次数: 0

A DNA concatemer-encoded CRISPR/Cas12a fluorescence sensor for sensitive detection of Pb2+ based on DNAzymes

IF 4.2 3区化学

Analyst Pub Date : 2025-03-26 DOI: 10.1039/d5an00189g

Shaoying He, Wei Lin, Xin Liu, Fei Li, Hong Liang, Huo Xu, Chunhua Lu, Chao Xing

{"title":"A DNA concatemer-encoded CRISPR/Cas12a fluorescence sensor for sensitive detection of Pb2+ based on DNAzymes","authors":"Shaoying He, Wei Lin, Xin Liu, Fei Li, Hong Liang, Huo Xu, Chunhua Lu, Chao Xing","doi":"10.1039/d5an00189g","DOIUrl":"https://doi.org/10.1039/d5an00189g","url":null,"abstract":"Lead pollution presents a significant threat to ecological systems and human health, underscoring the urgent need for highly sensitive detection methods. Herein, we introduce a novel DNA concatemer-encoded CRISPR/Cas12a fluorescence sensor (MDD-Cas12a) for sensitive detection of Pb2+ based on DNAzymes. To accomplish this, we designed a substrate strand containing a long DNA concatemer encoding multiple protospacer adjacent motifs (PAMs) and protospacer sequences for activation of the CRISPR/Cas12a system. The DNA concatemer was subsequently anchored to the surface of magnetic beads (MBs) to fabricate a MBs-DNA concatemer nanoprobe. In the presence of Pb2+, the DNAzyme structure catalyzes the cleavage of the substrate strand, leading to the release of DNA concatemers. Following magnetic separation, the released DNA concatemers significantly activate the non-specific trans-cleavage activity of the Cas12a/crRNA complex. The fluorescence reporter DNA is then completely cleaved by the activated Cas12a/crRNA complex, and the Pb2+ concentration in the sample can be quantified by measuring the fluorescence signal. By harnessing the specific recognition capability of DNAzymes for Pb2+, the programmability of DNA concatemers, and the self-amplification features of the CRISPR/Cas12a system, the MDD-Cas12a platform demonstrates high sensitivity and specificity for detecting Pb2+ in milk and lake water samples.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"68 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702966","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Development of a multiple reaction monitoring (MRM)-based LC-MS/MS method for the quantification of post-translational modifications on histone H3 variants in Arabidopsis thaliana†

IF 3.6 3区化学

Analyst Pub Date : 2025-03-25 DOI: 10.1039/D4AN01563K

Yajun Hu, Chenxi He, Lei Zhang and Hong Jin

{"title":"Development of a multiple reaction monitoring (MRM)-based LC-MS/MS method for the quantification of post-translational modifications on histone H3 variants in Arabidopsis thaliana†","authors":"Yajun Hu, Chenxi He, Lei Zhang and Hong Jin","doi":"10.1039/D4AN01563K","DOIUrl":"10.1039/D4AN01563K","url":null,"abstract":" Background: although the canonical histone H3.1 and its variant H3.3 differ by only four amino acids, they exhibit distinct genome-wide binding patterns and regulate different biological pathways. Post-translational modifications (PTMs) on histone tails mediate diverse downstream regulatory processes, raising the question of whether H3.1 and H3.3 harbor variant-specific modifications. However, the minimal amino acid differences between H3.1 and H3.3 make it challenging to distinguish and quantify them using traditional methods. Results: in this study, we developed an integrated multiple reaction monitoring (MRM)-based LC-MS/MS method to accurately differentiate and quantify K27 and K36 modifications on H3.1 and H3.3 in Arabidopsis thaliana. Our findings show that H3.1 contains more K27 methylation marks, associated with gene silencing, whereas H3.3 is enriched in K36 methylation, a mark of active transcription. Additionally, we compared K36 methylation levels in wild-type and SDG8-depleted cells, revealing that the K36 methyltransferase SDG8 shows a strong preference for H3.3 in both in vitro and in vivo assays. By analyzing public datasets, we further identified a strong correlation between H3.3 and the regions where H3K36me3 levels were reduced in sdg8 knockout cells. Significance: the MRM-based LC-MS/MS method established in this study provides a reliable and robust tool for the quantification of histone H3.1 and H3.3 PTMs in Arabidopsis thaliana. We demonstrate that the methyltransferase SDG8 shows a strong substrate preference for H3.3. This discovery highlights the importance of histone variant-specific modifications and suggests new avenues for research into their regulatory roles.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 8","pages":" 1688-1697"},"PeriodicalIF":3.6,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143695769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

LC-MS based untargeted metabolomics reveals benzoic acid as a predictive biomarker for embryo implantation potential

IF 4.2 3区化学

Analyst Pub Date : 2025-03-25 DOI: 10.1039/d4an01552e

Xi Chen, Ying Liu, Sheng-Nan Duan, Ping Wang, Yu-Nan Chen, Min Fu, Rong Liang, Xinxiang Zhang, Huan Shen, Ying-Lin Zhou, Cheng Shi

{"title":"LC-MS based untargeted metabolomics reveals benzoic acid as a predictive biomarker for embryo implantation potential","authors":"Xi Chen, Ying Liu, Sheng-Nan Duan, Ping Wang, Yu-Nan Chen, Min Fu, Rong Liang, Xinxiang Zhang, Huan Shen, Ying-Lin Zhou, Cheng Shi","doi":"10.1039/d4an01552e","DOIUrl":"https://doi.org/10.1039/d4an01552e","url":null,"abstract":"Evaluating the quality of embryos and implantation potential is a critical determinant of in vitro fertilization and embryo transfer, and it is also one of the main challenges of assisted reproductive technology. A reliable noninvasive method to choose the best candidate with real implantation potential for transfer from two day 3 embryos with equal morphological quality is still lacking clinically. In this article, a sensitive LC-MS method was developed and metabolomic profiling analysis in 3 days embryo culture medium was performed. Differential metabolites were analysed in two kinds of commercial culture media, and a total of common 66 metabolites were obtained from 106 independent samples in 5 batches. The relationship between changes in key metabolite benzoic acid concentrations and the embryo implantation result was discovered. This work improved coverage through conditional optimization, enhanced the reliability of omics data through multi batch validation, and provided a potential biomarker for evaluating the implantation potential of day 3 embryos.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"29 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143702969","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High-sensitivity differential scanning calorimetry using a MEMS thermopile chip for analyzing polymer crystallization

IF 4.2 3区化学

Analyst Pub Date : 2025-03-25 DOI: 10.1039/d5an00246j

Zechun Li, Shaokui Tan, Ming Li, Yuhang Yang, Haozhi Zhang, Xinxin Li, Pengcheng Xu

{"title":"High-sensitivity differential scanning calorimetry using a MEMS thermopile chip for analyzing polymer crystallization","authors":"Zechun Li, Shaokui Tan, Ming Li, Yuhang Yang, Haozhi Zhang, Xinxin Li, Pengcheng Xu","doi":"10.1039/d5an00246j","DOIUrl":"https://doi.org/10.1039/d5an00246j","url":null,"abstract":"This paper introduces a high-sensitivity differential scanning calorimetry (DSC) based on a MEMS single-crystalline silicon thermopile chip and its application for analyzing the crystallization process of Polyamide 6 (PA6) under various thermal processing conditions. The chip integrates 54 pairs of single-crystalline silicon thermocouples beneath a SiNx-suspended film, achieving a temperature responsivity of 31.5 mV/°C and a power responsivity of 147 V/W. Additionally, the chip’s cooling time constant is only 2.4 ms. The non-isothermal experimental results of PA6 suggest that melt-crystallization is suppressed at cooling rates exceeding the critical rate of 50°C/s, and cold-crystallization is suppressed at heating rates above the critical rate of 300°C/s. Thanks to its high sensitivity, this chip can detect subtle exothermic signals associated with the γ-α phase transition in PA6. The critical heating rate for this phase transition is determined to be 25°C/s. Isothermal experimental results show that PA6 undergoes crystallization within 70°C to 170°C, with the shortest half-crystallization time of ~1.1 s at 120°C. The high-sensitivity DSC technique proposed in this work holds great promise for studying the thermal behaviour of various materials under high heating and cooling rates.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"28 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143695770","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

IS-SCP: enhanced single-cell proteomics using an in situ simplified strategy†

IF 3.6 3区化学

Analyst Pub Date : 2025-03-24 DOI: 10.1039/D5AN00149H

Zhuo Yang, Yi-Rong Jiang, Qin-Qin Xu, Jian-Bo Chen, Jian-Zhang Pan, Xin Di and Qun Fang

{"title":"IS-SCP: enhanced single-cell proteomics using an in situ simplified strategy†","authors":"Zhuo Yang, Yi-Rong Jiang, Qin-Qin Xu, Jian-Bo Chen, Jian-Zhang Pan, Xin Di and Qun Fang","doi":"10.1039/D5AN00149H","DOIUrl":"10.1039/D5AN00149H","url":null,"abstract":"Advances in single-cell proteomics have enabled the investigation of the distinctive proteomic makeup of individual cells, significantly impacting biomedical research. However, most existing approaches involve complex sample preparation workflows and are sensitive to potential sample loss, which limits their applicability. In this paper, we reported an advanced workflow for easy-to-use single-cell proteome analysis using an in situ simplified strategy, named “in situ simplified single-cell proteomics (IS-SCP)”. This workflow was developed following a comprehensive evaluation of reagent mix, volume, and reaction conditions, notably including the utilization of a cleavable surfactant, n-decyl-disulfide-β-D-maltoside (DSSM). In comparison to previous workflows that require multiple steps in sample preparation, the IS-SCP workflow simplifies the single-cell proteome pretreatment to a single step of adding single-cell samples into a mixed reagent, which increases the repeatability and depth of single-cell proteome analysis. The IS-SCP workflow was applied to the proteomic analysis of single mammalian tumor cells, specifically HeLa and A549 cells, resulting in the quantification of an average of 3021 and 3289 protein groups, respectively. These results showed the potential of this workflow for investigating cellular heterogeneity at a deep single-cell level.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 8","pages":" 1679-1687"},"PeriodicalIF":3.6,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143677728","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Serum IgG galactosylation as a potential biomarker for diagnosis of echinococcosis

IF 4.2 3区化学

Analyst Pub Date : 2025-03-24 DOI: 10.1039/d4an01578a

Zhuoer Lu, Xiaoxiao Feng, Bin Fu, Xiaojin Mo, Ting Zhang, Liming Wei, Zhonghua Li, Haojie Lu

{"title":"Serum IgG galactosylation as a potential biomarker for diagnosis of echinococcosis","authors":"Zhuoer Lu, Xiaoxiao Feng, Bin Fu, Xiaojin Mo, Ting Zhang, Liming Wei, Zhonghua Li, Haojie Lu","doi":"10.1039/d4an01578a","DOIUrl":"https://doi.org/10.1039/d4an01578a","url":null,"abstract":"Echinococcosis is a serious and potentially fatal parasitic zoonosis, which can be divided into two subtypes in humans including cystic echinococcosis (CE) and alveolar echinococcosis (AE). It poses a great threat to patients’ lives, making timely diagnosis and subtype discrimination crucial. AE is easily confused with hepatocellular carcinoma (HCC) due to their highly similar features, so differential diagnosis is also imperative. In this work, the galactosylation level of serum IgG was analyzed by Matrix-Assisted Laser Desorption/Ionization Time of Flight Mass Spectrometry (MALDI-TOF-MS) in a cohort comprised of patients, including 100 diagnosed with CE, 27 with AE and 29 with HCC. The relative quantification of IgG digalactosyl (G2), monogalactosyl (G1), and agalactosylated (G0) N-glycans with the formula G0/(G1+G2*2) (IgG Gal-ratio) was obtained and found to effectively distinguish between echinococcosis patients and healthy controls, CE and AE, respectively. Meanwhile, the IgG Gal-ratio was evidently related to different types of CE (from CE1 to CE5) and the follow-up CE disease progress. Furthermore, the IgG Gal-ratio shown the potential differential diagnosis of AE and HCC. Thus, the results demonstrate that IgG Gal-ratio has the potential to be a biomarker for diagnosis and discrimination of echinococcosis, which also needs to be verified in further study.","PeriodicalId":63,"journal":{"name":"Analyst","volume":"94 1","pages":""},"PeriodicalIF":4.2,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143677880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

High performance charge detection mass spectrometry without ultra-high vacuum†

IF 3.6 3区化学

Analyst Pub Date : 2025-03-24 DOI: 10.1039/D5AN00019J

Emeline Hanozin, Conner C. Harper, Jacob S. Jordan, Zachary M. Miller and Evan R. Williams

{"title":"High performance charge detection mass spectrometry without ultra-high vacuum†","authors":"Emeline Hanozin, Conner C. Harper, Jacob S. Jordan, Zachary M. Miller and Evan R. Williams","doi":"10.1039/D5AN00019J","DOIUrl":"10.1039/D5AN00019J","url":null,"abstract":"Charge detection mass spectrometry (CDMS) measurements of individual ions using either Orbitrap or electrostatic ion trap-based instruments have heretofore been performed under ultra-high vacuum conditions (10−9 Torr or lower). The rationale for this expensive and often cumbersome requirement is that these measurements need to be performed in an environment where collisions with background gas do not adversely affect the measurements. Here, the use of an electrostatic trap that accepts a broad range of ion energies and a dynamic ion signal analysis method enables accurate CDMS mass measurements at pressures as high as 1 × 10−6 Torr, multiple orders of magnitude higher than previously demonstrated. Consistent, accurate masses were obtained for pentameric antibody complexes (∼800 kDa), adeno-associated viruses (∼4.8 MDa), and both ∼50 and ∼100 nm diameter polystyrene nanoparticles (∼35 MDa and ∼330 MDa, respectively) at pressures ranging from 1 × 10−8 Torr to 1 × 10−6 Torr. The relationships between ion mass, trap pressure, ion lifetimes, individual ion energies and survival rates were investigated over a 1 s trapping period. Larger ions are more robust to higher pressures. While the trapping lifetimes of smaller ions decrease with increasing pressure, enough survive long enough for accurate mass measurements to be made. Some ions are lost because collisional dampening decreases their energies below the minimum stability threshold of the trap, but others with sufficient energy are still lost due to collision-induced scattering that moves the ions too far from the central trapping axis.","PeriodicalId":63,"journal":{"name":"Analyst","volume":" 8","pages":" 1605-1616"},"PeriodicalIF":3.6,"publicationDate":"2025-03-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143677879","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Rapid and Sensitive Determination of Vancomycin by AIE-Active Fluorescent Probe for Clinical Monitoring

IF 4.2 3区化学

Analyst Pub Date : 2025-03-22 DOI: 10.1039/d4an01588f

Yige Lin, yujie Wang, Fang Fan, Guoyue Shi

引用次数: 0