The Argonaute (Ago) protein exhibits high specificity in nucleic acid recognition and cleavage, making it highly promising for biosensing applications. Its potential is further enhanced by its independence from protospacer adjacent motif (PAM) requirements and the cost-effectiveness of using short DNA guides. Both Ago and CRISPR/Cas systems face challenges in signal amplification, which limit their ability to detect targets at ultralow concentrations. To overcome this limitation, a thermostable quadratic amplification system (T-QAS) was constructed by integrating a thermostable nicking-enzyme-mediated amplification (NEMA) strategy with TtAgo. The system leverages the high stability of T-QAS at elevated temperatures to enhance guide–target interactions and decrease false positives caused by nonspecific amplification. Additionally, nanozyme is integrated with T-QAS to construct the AIESTA platform (all-in-one isothermal enzymatic signal transduction amplifier), which is a single-tube visual sensing platform. Within the AIESTA system, T-QAS improves specificity through high operational temperatures and offers programmable functions, enabling the sensitive detection of miRNA and foodborne toxins. The combination of T-QAS and nanozyme makes AIESTA a candidate of point-of-care testing (POCT) field, showcasing the potential for biosensing in resource-limited and complex environments.
{"title":"Programmable AIESTA: All-in-One Isothermal Enzymatic Signal Transduction Amplifier for Portable Profiling","authors":"Haoran Shen, Yanling Li, Kangling Tang, Hongzhi Liang, Zhen-Lin Xu, Yingju Liu, Weipeng Liu","doi":"10.1021/acs.analchem.5c00934","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00934","url":null,"abstract":"The Argonaute (Ago) protein exhibits high specificity in nucleic acid recognition and cleavage, making it highly promising for biosensing applications. Its potential is further enhanced by its independence from protospacer adjacent motif (PAM) requirements and the cost-effectiveness of using short DNA guides. Both Ago and CRISPR/Cas systems face challenges in signal amplification, which limit their ability to detect targets at ultralow concentrations. To overcome this limitation, a thermostable quadratic amplification system (T-QAS) was constructed by integrating a thermostable nicking-enzyme-mediated amplification (NEMA) strategy with TtAgo. The system leverages the high stability of T-QAS at elevated temperatures to enhance guide–target interactions and decrease false positives caused by nonspecific amplification. Additionally, nanozyme is integrated with T-QAS to construct the AIESTA platform (all-in-one isothermal enzymatic signal transduction amplifier), which is a single-tube visual sensing platform. Within the AIESTA system, T-QAS improves specificity through high operational temperatures and offers programmable functions, enabling the sensitive detection of miRNA and foodborne toxins. The combination of T-QAS and nanozyme makes AIESTA a candidate of point-of-care testing (POCT) field, showcasing the potential for biosensing in resource-limited and complex environments.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"36 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143737049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-31DOI: 10.1021/acs.analchem.4c04750
Ling Dai, Mengjun Deng, Kena Chen, Xueping Chen, Junjie Li
The in-frame internal tandem duplication of the FLT-3 gene (FLT3-ITD), a prevalent genetic aberration, significantly contributes to treatment failure and poor prognosis in acute myeloid leukemia (AML). A robust and cost-effective assay for minimal residual disease (MRD) detection in FLT3-ITD+ AML is crucial for guiding therapeutic decisions. However, current MRD monitoring methodologies for FLT3-ITD+ patients are limited by sensitivity and adaptability, particularly for dynamically quantifying complex and heterogeneous FLT3-ITD mutations. In this study, we developed a primer competition enhanced mutation accumulation (PCEMA) technique designed to selectively enrich FLT3-ITD in the context of abundant wild-type alleles. By integrating the PCEMA with capillary electrophoresis, we significantly improved the discrimination between mutant and wild-type genes, increasing the minimum detectable sensitivity to 0.001%, comparable to next-generation sequencing. The competitive amplification between ITD-specific and universal primers facilitated the selective enrichment of mutant alleles, enabling highly sensitive and specific real-time FLT3-ITD mutation monitoring. We thoroughly evaluated the analytical performance and adoptability of the PCEMA technique in conjunction with quantitative fluorescent PCR (qPCEMA). Our results demonstrated that qPCEMA quantitatively differentiates FLT3-ITD with a mutation frequency below 0.1%, offering an effective, rapid, and reliable method for long-term FLT3-ITD monitoring in clinical AML patients. The PCEMA technique, characterized by its robustness, sensitivity, specificity, timeliness, and adoptability, presents a promising alternative for clinical FLT3-ITD mutation detection. It is anticipated to provide significant technical support for timely diagnosis, prognosis assessment, drug evaluation, and personalized treatment of AML patients, with substantial potential for clinical application.
{"title":"Ultrasensitive Detection of FLT3-ITD Mutations via Primer Competition Enhanced Mutant Accumulation","authors":"Ling Dai, Mengjun Deng, Kena Chen, Xueping Chen, Junjie Li","doi":"10.1021/acs.analchem.4c04750","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04750","url":null,"abstract":"The in-frame internal tandem duplication of the FLT-3 gene (FLT3-ITD), a prevalent genetic aberration, significantly contributes to treatment failure and poor prognosis in acute myeloid leukemia (AML). A robust and cost-effective assay for minimal residual disease (MRD) detection in FLT3-ITD<sup>+</sup> AML is crucial for guiding therapeutic decisions. However, current MRD monitoring methodologies for FLT3-ITD<sup>+</sup> patients are limited by sensitivity and adaptability, particularly for dynamically quantifying complex and heterogeneous FLT3-ITD mutations. In this study, we developed a primer competition enhanced mutation accumulation (PCEMA) technique designed to selectively enrich FLT3-ITD in the context of abundant wild-type alleles. By integrating the PCEMA with capillary electrophoresis, we significantly improved the discrimination between mutant and wild-type genes, increasing the minimum detectable sensitivity to 0.001%, comparable to next-generation sequencing. The competitive amplification between ITD-specific and universal primers facilitated the selective enrichment of mutant alleles, enabling highly sensitive and specific real-time FLT3-ITD mutation monitoring. We thoroughly evaluated the analytical performance and adoptability of the PCEMA technique in conjunction with quantitative fluorescent PCR (qPCEMA). Our results demonstrated that qPCEMA quantitatively differentiates FLT3-ITD with a mutation frequency below 0.1%, offering an effective, rapid, and reliable method for long-term FLT3-ITD monitoring in clinical AML patients. The PCEMA technique, characterized by its robustness, sensitivity, specificity, timeliness, and adoptability, presents a promising alternative for clinical FLT3-ITD mutation detection. It is anticipated to provide significant technical support for timely diagnosis, prognosis assessment, drug evaluation, and personalized treatment of AML patients, with substantial potential for clinical application.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"58 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143737044","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-31DOI: 10.1021/acs.analchem.5c01608
Laura N. Taylor, Lisa A. Holland, Makenzie T. Witzel
Corrections are reported for the Supporting Information in our published article reflecting two peak areas which were mistyped, revised data displaying only significant digits in means and standard deviations, and an amended computation in Table S11A. The revised Supporting Information file is given here. Also, corrections are reported for the published article reflecting two values. In the abstract, on page 5082 and in Figure 5A: For 1918 H1N1 (A/Brevig Mission/1/18) neuraminidase, the inhibition constant of the transition state analog 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (DANA) is 3.4 μM, which is derived from the corrected value of 3.44 ± 0.76. On page 5080: The H1N1 % enzyme activity remaining in the presence of sodium chloride has a standard deviation of ± 1. The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.5c01608. Electropherograms and data from experiments associated with Ki measurements and method development (PDF) Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html. This article has not yet been cited by other publications.
{"title":"Correction to “Native Capillary Nanogel Electrophoresis Assay of Inhibitors of Neuraminidases Derived from H1N1 and H5N1 Influenza A Pandemics”","authors":"Laura N. Taylor, Lisa A. Holland, Makenzie T. Witzel","doi":"10.1021/acs.analchem.5c01608","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01608","url":null,"abstract":"Corrections are reported for the Supporting Information in our published article reflecting two peak areas which were mistyped, revised data displaying only significant digits in means and standard deviations, and an amended computation in Table S11A. The revised Supporting Information file is given here. Also, corrections are reported for the published article reflecting two values. In the abstract, on page 5082 and in Figure 5A: For 1918 H1N1 (A/Brevig Mission/1/18) neuraminidase, the inhibition constant of the transition state analog 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (DANA) is 3.4 μM, which is derived from the corrected value of 3.4<sub>4</sub> ± 0.7<sub>6</sub>. On page 5080: The H1N1 % enzyme activity remaining in the presence of sodium chloride has a standard deviation of ± 1. The Supporting Information is available free of charge at https://pubs.acs.org/doi/10.1021/acs.analchem.5c01608. Electropherograms and data from experiments associated with <i>K</i><sub><i>i</i></sub> measurements and method development (PDF) Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system: http://pubs.acs.org/page/copyright/permissions.html. This article has not yet been cited by other publications.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"18 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143737050","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-30DOI: 10.1021/acs.analchem.4c06869
Qiwen Liu, Rentao Tang, Xiyu Chen, Jiamei Chen, Yang Huang, Sheng Wang, Ning Gan, Shengfeng Huang
The development of highly sensitive detection methods for bioanalysis is crucial for early disease diagnosis. Electrochemical biosensing technology offers unique advantages in this area due to its rapid response, high sensitivity, and low cost. However, achieving efficient and rapid transfer of signaling molecules to the electrode interface to facilitate effective interaction between signal molecules and the sensing surface remains a critical challenge for ultrasensitive electrochemical detection. In this study, we discovered that single-stranded DNA-modified magnetic nanoprobes (signal probe A) subjected to cryogenic treatment can rapidly form an orderly monolayer at the electrode interface under an external magnetic field, while this phenomenon was not observed with double-stranded DNA-modified magnetic nanoprobes (signal probe B). Building on this finding, we developed a signal probe with a protective complementary strand (signal probe B) that, upon interaction with target molecules, is converted into signal probe A. This transformation, combined with cryogenic treatment, enables the ultrasensitive detection of target molecules. Using miRNA-21 and a carcinoembryonic antigen (CEA) as model targets, we optimized the detection conditions, achieving a detection limit as low as 3.4 aM for miRNA-21 and 0.28 fg/mL for CEA with excellent versatility. In summary, this study introduces a highly efficient, rapid, enzyme-free, and environmentally friendly electrochemical signal amplification strategy. This approach not only provides an innovative solution for the ultrasensitive bioanalysis but also offers new insights into enhancing signal molecule–sensor interface interactions in electrochemical biosensors.
{"title":"Cryogenically Induced Highly Ordered Single-Strand DNA-Modified Magnetic Nanoprobes for Rapid and Ultrasensitive Bioanalysis","authors":"Qiwen Liu, Rentao Tang, Xiyu Chen, Jiamei Chen, Yang Huang, Sheng Wang, Ning Gan, Shengfeng Huang","doi":"10.1021/acs.analchem.4c06869","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06869","url":null,"abstract":"The development of highly sensitive detection methods for bioanalysis is crucial for early disease diagnosis. Electrochemical biosensing technology offers unique advantages in this area due to its rapid response, high sensitivity, and low cost. However, achieving efficient and rapid transfer of signaling molecules to the electrode interface to facilitate effective interaction between signal molecules and the sensing surface remains a critical challenge for ultrasensitive electrochemical detection. In this study, we discovered that single-stranded DNA-modified magnetic nanoprobes (signal probe A) subjected to cryogenic treatment can rapidly form an orderly monolayer at the electrode interface under an external magnetic field, while this phenomenon was not observed with double-stranded DNA-modified magnetic nanoprobes (signal probe B). Building on this finding, we developed a signal probe with a protective complementary strand (signal probe B) that, upon interaction with target molecules, is converted into signal probe A. This transformation, combined with cryogenic treatment, enables the ultrasensitive detection of target molecules. Using miRNA-21 and a carcinoembryonic antigen (CEA) as model targets, we optimized the detection conditions, achieving a detection limit as low as 3.4 aM for miRNA-21 and 0.28 fg/mL for CEA with excellent versatility. In summary, this study introduces a highly efficient, rapid, enzyme-free, and environmentally friendly electrochemical signal amplification strategy. This approach not only provides an innovative solution for the ultrasensitive bioanalysis but also offers new insights into enhancing signal molecule–sensor interface interactions in electrochemical biosensors.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"58 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143737051","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Accurately diagnosing infectious diseases in a resource-limited setting is a major challenge. Plasmonic materials, via localized surface plasmon resonance (LSPR), have greatly enhanced fluorescence signal and detection sensitivity. However, traditional plasmonic-enhanced fluorescence methods largely rely on near-infrared or visible-light fluorophores with small Stokes shift, limiting naked-eye visibility without filters. In this study, we developed a novel plasmonic silver film (pSilverF) to enhance visible-light fluorescence with large Stokes shift, allowing for improved biomarker detection sensitivity under naked-eye observation. Integrated with bright fluorescent nanoparticles, we designed a multiplexed assay for detecting Hepatitis C Virus (HCV), Hepatitis B Virus (HBV), and Human Immunodeficiency Virus (HIV) antibodies, achieving detection sensitivities down to 0.0032, 0.023, and 0.168 NCU/mL, respectively. In a cohort of 68 clinical samples, our method achieved 100% sensitivity and specificity for HIV and HCV detection and 96% sensitivity and 100% specificity for HBV detection. Notably, the results can be visualized by the naked eye and directly captured by a standard mobile phone camera without any modification for signal analysis using RGB image splitting. This platform demonstrated potential for field detection of multiple infectious diseases with simple settings, providing a useful tool for disease control in communities and areas with limited medical resources.
{"title":"Field Detection of Multiple Infectious Diseases with Naked Eye Using Plasmonic-Enhanced Fluorescent Nanoparticles","authors":"Xuan Tan, Jingkai Yang, Ying Liu, Zexi Tang, HongJun Xiao, Jiahui Lv, Yun He, Ruibin Hu, Ziqi Jin, Shiyu Chen, Ziyi Xu, Li Cheng, Jiaxin Li, Rongrong Zou, Xiaohe Li, Panlin Shao, Jing Yuan, Bo Zhang","doi":"10.1021/acs.analchem.4c07106","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c07106","url":null,"abstract":"Accurately diagnosing infectious diseases in a resource-limited setting is a major challenge. Plasmonic materials, via localized surface plasmon resonance (LSPR), have greatly enhanced fluorescence signal and detection sensitivity. However, traditional plasmonic-enhanced fluorescence methods largely rely on near-infrared or visible-light fluorophores with small Stokes shift, limiting naked-eye visibility without filters. In this study, we developed a novel plasmonic silver film (pSilverF) to enhance visible-light fluorescence with large Stokes shift, allowing for improved biomarker detection sensitivity under naked-eye observation. Integrated with bright fluorescent nanoparticles, we designed a multiplexed assay for detecting Hepatitis C Virus (HCV), Hepatitis B Virus (HBV), and Human Immunodeficiency Virus (HIV) antibodies, achieving detection sensitivities down to 0.0032, 0.023, and 0.168 NCU/mL, respectively. In a cohort of 68 clinical samples, our method achieved 100% sensitivity and specificity for HIV and HCV detection and 96% sensitivity and 100% specificity for HBV detection. Notably, the results can be visualized by the naked eye and directly captured by a standard mobile phone camera without any modification for signal analysis using RGB image splitting. This platform demonstrated potential for field detection of multiple infectious diseases with simple settings, providing a useful tool for disease control in communities and areas with limited medical resources.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"72 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143737052","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-30DOI: 10.1021/acs.analchem.5c01163
Ting Huang, Naihan Huang, Qianying Zhou, Baoping Xie, Wenjun Duan, Bin Sun, Zong Dai, Jun Chen, Jin-Xiang Chen
The highly sensitive and rapid imaging of miRNA in living cells promises to advance our understanding of diseases and promote their diagnosis and treatment. To enhance the sensitivity of miRNA imaging, a series of cascade signal amplification strategies based on enzyme-free methods have been developed. However, these cascaded amplification strategies involve complex designs and multiple amplification mechanisms, leading to potential side effects. Herein, we have developed a novel hairpins@ZIF-8 nanosystem by rationally integrating ZIF-8 with a cascaded self-feedback DNAzyme circuit (CSDC), achieving highly sensitive and rapid imaging of miRNA in living cells. ZIF-8 facilitates the efficient transfection of nucleic acid probes into cells and provides the necessary cofactor ions for CSDC. The developed CSDC possesses exponential amplification based solely on the DNAzyme mechanism. Benefiting from the exponential amplification efficiency, the CSDC exhibited higher sensitivity than traditional DNAzyme-based amplification, with a detection limit of 2.28 fM, approximately 105 times more sensitive than traditional DNAzyme-based amplification. This hairpins@ZIF-8 nanosystem demonstrated strong practical application capabilities, effectively reflecting fluctuations in intracellular miRNA levels and successfully distinguishing between normal and tumor cells based on miRNA expression differences. It could also be applied for in vivo miRNA imaging. This proposed strategy is anticipated to pave the way for innovative amplification approaches and serve as a vital instrument in miRNA-related research, diagnosis, and treatment.
{"title":"Target-Triggered Cascaded Self-Feedback DNAzyme Circuit Loaded in ZIF-8 for Highly Sensitive miRNA Imaging in Living Cells","authors":"Ting Huang, Naihan Huang, Qianying Zhou, Baoping Xie, Wenjun Duan, Bin Sun, Zong Dai, Jun Chen, Jin-Xiang Chen","doi":"10.1021/acs.analchem.5c01163","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c01163","url":null,"abstract":"The highly sensitive and rapid imaging of miRNA in living cells promises to advance our understanding of diseases and promote their diagnosis and treatment. To enhance the sensitivity of miRNA imaging, a series of cascade signal amplification strategies based on enzyme-free methods have been developed. However, these cascaded amplification strategies involve complex designs and multiple amplification mechanisms, leading to potential side effects. Herein, we have developed a novel hairpins@ZIF-8 nanosystem by rationally integrating ZIF-8 with a cascaded self-feedback DNAzyme circuit (CSDC), achieving highly sensitive and rapid imaging of miRNA in living cells. ZIF-8 facilitates the efficient transfection of nucleic acid probes into cells and provides the necessary cofactor ions for CSDC. The developed CSDC possesses exponential amplification based solely on the DNAzyme mechanism. Benefiting from the exponential amplification efficiency, the CSDC exhibited higher sensitivity than traditional DNAzyme-based amplification, with a detection limit of 2.28 fM, approximately 10<sup>5</sup> times more sensitive than traditional DNAzyme-based amplification. This hairpins@ZIF-8 nanosystem demonstrated strong practical application capabilities, effectively reflecting fluctuations in intracellular miRNA levels and successfully distinguishing between normal and tumor cells based on miRNA expression differences. It could also be applied for in vivo miRNA imaging. This proposed strategy is anticipated to pave the way for innovative amplification approaches and serve as a vital instrument in miRNA-related research, diagnosis, and treatment.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"72 3 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143737053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-29DOI: 10.1021/acs.analchem.5c00807
Jie Gao, Yucun Zhang, Rui Zhu, Mu Li, Fei Xie, Changyin Li, Bingqian Li, Yungang Zhang
Carbon disulfide (CS2) and sulfur dioxide (SO2) are typical indicative gases for the early warning and diagnosis of faults in gas-insulated switchgear. In this study, an optical sensing system is reported for online detection of CS2 and SO2 based on a bidirectional self-correction technology (BSCT). First, the differential absorption signals of CS2 and SO2 in the wavelength range of 195–230 nm are obtained using the UV differential optical absorption spectroscopy (UV-DOAS) technique. On this basis, a BSCT is proposed to decouple the spectral lines that exhibit significant overlap. This method primarily employs bidirectional difference spectroscopy to mutually correct the spectra of CS2 and SO2, combined with spectral reconstruction to extract the single-component absorption signals of CS2 and SO2 from the mixed gas spectra. Furthermore, the effectiveness of this novel decoupling technique is validated by comparing the decoupling results with the absorption spectra of single-component standard gases at corresponding concentrations. Finally, the quantitative relationships between the concentrations of target gases (CS2, SO2) and the optical parameters are modeled using the least-squares method. The experimental results show that the mean absolute percentage errors of CS2 (19.00–3735.35 ppb) and SO2 (0.19–38.77 ppm) are 0.543 and 0.521%, respectively. At an effective optical range of 50 cm, the system achieves the lowest detection limits of 0.5 ppb for CS2 and 12 ppb for SO2, representing the best results reported to date for the online detection of CS2 and SO2 in the ppb-ppm range.
{"title":"Optical Sensing System Based on Bidirectional Self-Correction Technology: An Online Detection Method for Carbon Disulfide and Sulfur Dioxide in Gas-Insulated Switchgear","authors":"Jie Gao, Yucun Zhang, Rui Zhu, Mu Li, Fei Xie, Changyin Li, Bingqian Li, Yungang Zhang","doi":"10.1021/acs.analchem.5c00807","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00807","url":null,"abstract":"Carbon disulfide (CS<sub>2</sub>) and sulfur dioxide (SO<sub>2</sub>) are typical indicative gases for the early warning and diagnosis of faults in gas-insulated switchgear. In this study, an optical sensing system is reported for online detection of CS<sub>2</sub> and SO<sub>2</sub> based on a bidirectional self-correction technology (BSCT). First, the differential absorption signals of CS<sub>2</sub> and SO<sub>2</sub> in the wavelength range of 195–230 nm are obtained using the UV differential optical absorption spectroscopy (UV-DOAS) technique. On this basis, a BSCT is proposed to decouple the spectral lines that exhibit significant overlap. This method primarily employs bidirectional difference spectroscopy to mutually correct the spectra of CS<sub>2</sub> and SO<sub>2</sub>, combined with spectral reconstruction to extract the single-component absorption signals of CS<sub>2</sub> and SO<sub>2</sub> from the mixed gas spectra. Furthermore, the effectiveness of this novel decoupling technique is validated by comparing the decoupling results with the absorption spectra of single-component standard gases at corresponding concentrations. Finally, the quantitative relationships between the concentrations of target gases (CS<sub>2</sub>, SO<sub>2</sub>) and the optical parameters are modeled using the least-squares method. The experimental results show that the mean absolute percentage errors of CS<sub>2</sub> (19.00–3735.35 ppb) and SO<sub>2</sub> (0.19–38.77 ppm) are 0.543 and 0.521%, respectively. At an effective optical range of 50 cm, the system achieves the lowest detection limits of 0.5 ppb for CS<sub>2</sub> and 12 ppb for SO<sub>2</sub>, representing the best results reported to date for the online detection of CS<sub>2</sub> and SO<sub>2</sub> in the ppb-ppm range.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"72 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734224","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Heparin is a vital macromolecule that regulates blood coagulation, while protamine is an essential polypeptide clinically used to counteract heparin overdose. Detecting both heparin and its antidote protamine under physiological conditions is crucial for biological and clinical applications. This report introduces a cucurbituril[8] (CB[8])-based phosphorescent probe for their detection. The method employs a nanoassembly induced phosphorescence switch-on mechanism for heparin sensing and a disassembly induced phosphorescence switch-off approach for protamine detection. An arginine-rich guest forms a supramolecular complex with heparin, enhancing phosphorescence under secondary confinement and enabling its detection. Conversely, protamine sulfate, as a stronger competitor for heparin, disrupts the probe-heparin aggregates, leading to emission quenching and protamine sensing. This sensor demonstrated high selectivity in detecting both analytes in biological samples, such as human blood serum and urine. The detection limits for heparin and protamine were determined to be 61 and 82 ng/mL in 10% HBS, respectively.
{"title":"An Aggregation-Induced Room Temperature Phosphorescence Probe for the Efficient and Selective Detection of Heparin and Protamine","authors":"Priyam Das, Sampurna Routray, Malay Kumar Baroi, Tanushree Das, Debapratim Das","doi":"10.1021/acs.analchem.5c00382","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00382","url":null,"abstract":"Heparin is a vital macromolecule that regulates blood coagulation, while protamine is an essential polypeptide clinically used to counteract heparin overdose. Detecting both heparin and its antidote protamine under physiological conditions is crucial for biological and clinical applications. This report introduces a cucurbituril[8] (CB[8])-based phosphorescent probe for their detection. The method employs a nanoassembly induced phosphorescence switch-on mechanism for heparin sensing and a disassembly induced phosphorescence switch-off approach for protamine detection. An arginine-rich guest forms a supramolecular complex with heparin, enhancing phosphorescence under secondary confinement and enabling its detection. Conversely, protamine sulfate, as a stronger competitor for heparin, disrupts the probe-heparin aggregates, leading to emission quenching and protamine sensing. This sensor demonstrated high selectivity in detecting both analytes in biological samples, such as human blood serum and urine. The detection limits for heparin and protamine were determined to be 61 and 82 ng/mL in 10% HBS, respectively.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"20 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143736331","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-29DOI: 10.1021/acs.analchem.5c00711
Juan Song, Yue Zheng, Xiaodan Huang, Xiyuan Yu, Rui Su, Zhi Zhu, Chaoyong Yang, Zengpeng Li, Yu Jiang, Huimin Zhang
Multivalent strategies have been widely applied in the microfluidic interface to boost the capture efficiency of target cells. However, achieving a balance between binding kinetics and thermodynamics in existing multivalent affinity interfaces remains challenging. Here, we propose a synergistic Aptamer-nanobody hetero-Multivalency Programmable magnetic fluid microfluidic chip (AMP-chip) which utilizes the combined advantages of ligands to enhance both thermodynamic and kinetic properties of the capture interface. The AMP-chip integrates two distinct noninterfering recognition molecules: one with high affinity and another with rapid binding capability, both of which are assembled onto nanomagnetic beads. This integration achieves intermolecular complementarity, effectively enhancing the binding kinetics and thermodynamic stability. We chose mutually noninterfering CD71 recognition targets, a high-affinity nanobody (NB) and a rapid-binding aptamer (XQ 2d), and fully utilized the respective advantages of these ligands to facilitate rapid and tight recognition of the CD71 receptor on target cells. By integrating a herringbone microarray into an AMP-chip to further increase the cell–ligand interaction, we significantly improved the sensitivity and accuracy of circulating nucleated red blood cell (cNRBC) isolation from the peripheral blood mononuclear cells (PBMCs) of pregnant women. Additionally, the ligands were primarily fixed to the chip by magnetic force without chemical bonding, enabling nondestructive cell release and preserving high cell viability for subsequent molecular analyses. Overall, this approach offers a novel thermodynamic–kinetic synergistic heteromultivalency interface with significant potential for clinical applications.
{"title":"Enhancing Thermodynamic and Kinetic Performance of Microfluidic Interface-Based Circulating Fetal Cell Isolation for Noninvasive Prenatal Testing","authors":"Juan Song, Yue Zheng, Xiaodan Huang, Xiyuan Yu, Rui Su, Zhi Zhu, Chaoyong Yang, Zengpeng Li, Yu Jiang, Huimin Zhang","doi":"10.1021/acs.analchem.5c00711","DOIUrl":"https://doi.org/10.1021/acs.analchem.5c00711","url":null,"abstract":"Multivalent strategies have been widely applied in the microfluidic interface to boost the capture efficiency of target cells. However, achieving a balance between binding kinetics and thermodynamics in existing multivalent affinity interfaces remains challenging. Here, we propose a synergistic <b>A</b>ptamer-nanobody hetero-<b>M</b>ultivalency <b>P</b>rogrammable magnetic fluid microfluidic chip (<b>AMP-chip</b>) which utilizes the combined advantages of ligands to enhance both thermodynamic and kinetic properties of the capture interface. The AMP-chip integrates two distinct noninterfering recognition molecules: one with high affinity and another with rapid binding capability, both of which are assembled onto nanomagnetic beads. This integration achieves intermolecular complementarity, effectively enhancing the binding kinetics and thermodynamic stability. We chose mutually noninterfering CD71 recognition targets, a high-affinity nanobody (NB) and a rapid-binding aptamer (XQ 2d), and fully utilized the respective advantages of these ligands to facilitate rapid and tight recognition of the CD71 receptor on target cells. By integrating a herringbone microarray into an AMP-chip to further increase the cell–ligand interaction, we significantly improved the sensitivity and accuracy of circulating nucleated red blood cell (cNRBC) isolation from the peripheral blood mononuclear cells (PBMCs) of pregnant women. Additionally, the ligands were primarily fixed to the chip by magnetic force without chemical bonding, enabling nondestructive cell release and preserving high cell viability for subsequent molecular analyses. Overall, this approach offers a novel thermodynamic–kinetic synergistic heteromultivalency interface with significant potential for clinical applications.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"183 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143734223","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-03-28DOI: 10.1021/acs.analchem.4c06496
Ondřej Peterka, Yasmin Kadyrbekova, Robert Jirásko, Zuzana Lásko, Bohuslav Melichar, Michal Holčapek
Chemical derivatization involves the reaction of an analyte with a derivatization agent to modify its structure, improving the peak shape, chromatographic performance, structural analysis, ionization efficiency, and sensitivity. A novel derivatization method using 3-(chlorosulfonyl)benzoic acid is developed for the determination of monoacylglycerols, diacylglycerols, free sterols, and tocopherols using the reversed-phase ultra-high-performance liquid chromatography–tandem mass spectrometry (RP-UHPLC/MS/MS) method in the negative ion mode. The chromatographic and mass spectrometric properties of derivatized lipids are investigated by using 29 lipid standards spanning four lipid classes. The derivatization process is optimized using pooled plasma spiked by 9 internal standards, achieving an optimal yield with a reaction time of 40 min at 60 °C. The stability of the derivatives is confirmed, with short-term stability maintained for 10 h at 4 °C and long-term stability preserved for 5 days at −80 °C. The repeatability and reproducibility are verified by one/two operator(s), which underscores the simplicity and robustness of the method, and calibration curves with high linear regression coefficients illustrate the accuracy of the method. The derivatization approach, which combines RP-UHPLC/MS/MS and the use of specific fragmentation patterns, significantly reduces limits of detection, reaching 15–25 pmol/mL for free sterols in plasma. The optimized method is applied to the analysis of human plasma, leading to the identification of 92 lipid species in the targeted lipid classes. This represents a substantial improvement in sensitivity and detection capabilities compared to those of previously reported methods.
{"title":"Novel Charge-Switch Derivatization Method Using 3-(Chlorosulfonyl)benzoic Acid for Sensitive RP-UHPLC/MS/MS Analysis of Acylglycerols, Sterols, and Prenols","authors":"Ondřej Peterka, Yasmin Kadyrbekova, Robert Jirásko, Zuzana Lásko, Bohuslav Melichar, Michal Holčapek","doi":"10.1021/acs.analchem.4c06496","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06496","url":null,"abstract":"Chemical derivatization involves the reaction of an analyte with a derivatization agent to modify its structure, improving the peak shape, chromatographic performance, structural analysis, ionization efficiency, and sensitivity. A novel derivatization method using 3-(chlorosulfonyl)benzoic acid is developed for the determination of monoacylglycerols, diacylglycerols, free sterols, and tocopherols using the reversed-phase ultra-high-performance liquid chromatography–tandem mass spectrometry (RP-UHPLC/MS/MS) method in the negative ion mode. The chromatographic and mass spectrometric properties of derivatized lipids are investigated by using 29 lipid standards spanning four lipid classes. The derivatization process is optimized using pooled plasma spiked by 9 internal standards, achieving an optimal yield with a reaction time of 40 min at 60 °C. The stability of the derivatives is confirmed, with short-term stability maintained for 10 h at 4 °C and long-term stability preserved for 5 days at −80 °C. The repeatability and reproducibility are verified by one/two operator(s), which underscores the simplicity and robustness of the method, and calibration curves with high linear regression coefficients illustrate the accuracy of the method. The derivatization approach, which combines RP-UHPLC/MS/MS and the use of specific fragmentation patterns, significantly reduces limits of detection, reaching 15–25 pmol/mL for free sterols in plasma. The optimized method is applied to the analysis of human plasma, leading to the identification of 92 lipid species in the targeted lipid classes. This represents a substantial improvement in sensitivity and detection capabilities compared to those of previously reported methods.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"183 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-03-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143723758","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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