Pub Date : 2025-01-10DOI: 10.1021/acs.analchem.4c04794
Jordan S. Stanberry, Hunter B. Andrews, Cyril V. Thompson, Brian W. Ticknor, Benjamin T. Manard
A novel employment of single particle-inductively coupled plasma-mass spectrometry (SP-ICP-MS) was developed, where a microextraction (ME) probe is used to sample nanoparticles from a surface and analyze them in a single analytical step. The effects of several parameters on the performance of ME-SP-ICP-MS were investigated, including the flow rate, choice of carrier solution, particle size, and the design of the microextraction probe head itself. The optimized ME-SP-ICP-MS technique was used to compare the extraction efficiency (EE, defined as the ratio of particles measured to particles deposited on the surface) of the commercial probe head to a newly designed SP polyether ether ketone (PEEK) probe head. The SP PEEK probe head was found to have increased EE compared to the commercial probe head (8.5 ± 3% vs 3.9 ± 3%, respectively). Increasing the carrier solution flow rate was found to decrease the total analysis time at the cost of decreasing EE. Extraction efficiencies for ME-SP-ICP-MS were typically 4–10%, which is similar to transport efficiencies (1–10%) for conventional SP-ICP-MS. Lastly, ME-SP-ICP-MS was employed for the analysis of nano- and microparticles. The sizes of gold nanoparticles, 30 ± 3 and 51 ± 1.9 nm (certified sizes), and iron-based microparticles, 1000 ± 50 nm (certified size), were accurately determined to be 32.2 ± 2.5, 50.8 ± 3.4, and 1030 ± 57 nm, respectively, by ME-SP-ICP-MS. This work demonstrates the potential of ME-SP-ICP-MS for the direct analysis of particles on common collection surfaces (GSR tabs, carbon planchettes, etc.) while retaining spatial information on particle distribution across the surface.
{"title":"Microextraction-Single Particle-Inductively Coupled Plasma-Mass Spectrometry for the Direct Analysis of Nanoparticles on Surfaces","authors":"Jordan S. Stanberry, Hunter B. Andrews, Cyril V. Thompson, Brian W. Ticknor, Benjamin T. Manard","doi":"10.1021/acs.analchem.4c04794","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04794","url":null,"abstract":"A novel employment of single particle-inductively coupled plasma-mass spectrometry (SP-ICP-MS) was developed, where a microextraction (ME) probe is used to sample nanoparticles from a surface and analyze them in a single analytical step. The effects of several parameters on the performance of ME-SP-ICP-MS were investigated, including the flow rate, choice of carrier solution, particle size, and the design of the microextraction probe head itself. The optimized ME-SP-ICP-MS technique was used to compare the extraction efficiency (EE, defined as the ratio of particles measured to particles deposited on the surface) of the commercial probe head to a newly designed SP polyether ether ketone (PEEK) probe head. The SP PEEK probe head was found to have increased EE compared to the commercial probe head (8.5 ± 3% vs 3.9 ± 3%, respectively). Increasing the carrier solution flow rate was found to decrease the total analysis time at the cost of decreasing EE. Extraction efficiencies for ME-SP-ICP-MS were typically 4–10%, which is similar to transport efficiencies (1–10%) for conventional SP-ICP-MS. Lastly, ME-SP-ICP-MS was employed for the analysis of nano- and microparticles. The sizes of gold nanoparticles, 30 ± 3 and 51 ± 1.9 nm (certified sizes), and iron-based microparticles, 1000 ± 50 nm (certified size), were accurately determined to be 32.2 ± 2.5, 50.8 ± 3.4, and 1030 ± 57 nm, respectively, by ME-SP-ICP-MS. This work demonstrates the potential of ME-SP-ICP-MS for the direct analysis of particles on common collection surfaces (GSR tabs, carbon planchettes, etc.) while retaining spatial information on particle distribution across the surface.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"28 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940544","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c05998
Liu-Na Wei, Lin Luo, Bei Li, Qing-Chun Yin, Hongtao Lei, Bing-Zhi Wang, Tian Guan, Zhen-Lin Xu
The rapid, sensitive, and accurate detection of paralytic shellfish toxins (PSTs), such as saxitoxin (STX), is critical for protecting human health due to the frequent occurrence of toxic red tides. In this work, to address the low affinity of traditional mouse monoclonal antibodies (m-mAbs), rabbit monoclonal antibodies (r-mAbs) against STX were produced by a single B-cell sorting culture and a cross-selection strategy. The r-mAbs showed 100-fold improvement in sensitivity (IC50 = 0.018 ng/mL), improved stability under strong acid/alkali resistance (pH = 3–12), and high tolerance to organic solvent (MOH, 30%) in comparison with m-mAbs. The structural analysis of PSTs shows that the r-mAbs primarily interacted with common functional groups in PSTs, including guanidine, amide, and alcoholic hydroxyl groups (−OH). R-mAbs3C3 recognized STX, GTX2/3, C1/C2, NEO, and dcSTX with broad specificity through hydrogen bonding facilitated by Asp93, Gln27, Ser27A, and Asp95 in the complementarity-determining region (CDR) pocket. In contrast, r-mAbs3C7 displayed high specificity for STX, mediated by Ser98, Leu96, Ser30, Tyr97, and Ser53, which formed hydrogen bonds and hydrophobic interactions. The lack of interaction with Asp93 and Asp95 in r-mAbs3C7 likely contributed to its enhanced specificity. The anti-STX r-mAbs3C3 was then used to develop an ultrasensitive lateral flow immunoassay (LFIA) for the detection of STX with a cutoff value of 100 pg/mL. The LFIA demonstrated recovery rates ranging from 86.6 to 106.3% in food samples (mussels, nassariidae, bullacta exarata, and fresh water). Given its high target selectivity and sensitivity, this study provides a sight for rapid, sensitive, and accurate monitoring of food toxins.
{"title":"Ultrasensitive “Hunter of Target”: Rabbit Monoclonal Antibody-Based Competitive Lateral Flow Immunoassay for Saxitoxin","authors":"Liu-Na Wei, Lin Luo, Bei Li, Qing-Chun Yin, Hongtao Lei, Bing-Zhi Wang, Tian Guan, Zhen-Lin Xu","doi":"10.1021/acs.analchem.4c05998","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05998","url":null,"abstract":"The rapid, sensitive, and accurate detection of paralytic shellfish toxins (PSTs), such as saxitoxin (STX), is critical for protecting human health due to the frequent occurrence of toxic red tides. In this work, to address the low affinity of traditional mouse monoclonal antibodies (m-mAbs), rabbit monoclonal antibodies (r-mAbs) against STX were produced by a single B-cell sorting culture and a cross-selection strategy. The r-mAbs showed 100-fold improvement in sensitivity (IC<sub>50</sub> = 0.018 ng/mL), improved stability under strong acid/alkali resistance (pH = 3–12), and high tolerance to organic solvent (MOH, 30%) in comparison with m-mAbs. The structural analysis of PSTs shows that the r-mAbs primarily interacted with common functional groups in PSTs, including guanidine, amide, and alcoholic hydroxyl groups (−OH). R-mAbs3C3 recognized STX, GTX2/3, C1/C2, NEO, and dcSTX with broad specificity through hydrogen bonding facilitated by Asp93, Gln27, Ser27A, and Asp95 in the complementarity-determining region (CDR) pocket. In contrast, r-mAbs3C7 displayed high specificity for STX, mediated by Ser98, Leu96, Ser30, Tyr97, and Ser53, which formed hydrogen bonds and hydrophobic interactions. The lack of interaction with Asp93 and Asp95 in r-mAbs3C7 likely contributed to its enhanced specificity. The anti-STX r-mAbs3C3 was then used to develop an ultrasensitive lateral flow immunoassay (LFIA) for the detection of STX with a cutoff value of 100 pg/mL. The LFIA demonstrated recovery rates ranging from 86.6 to 106.3% in food samples (mussels, nassariidae, bullacta exarata, and fresh water). Given its high target selectivity and sensitivity, this study provides a sight for rapid, sensitive, and accurate monitoring of food toxins.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"25 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940534","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c05892
Junqiang Fu, Dongquan Leng, Jingui Chen, Xiang Ren, Yamei Li, Tingting Wu, Qin Wei, Bin Cai
A novel dual-mode microfluidic sensing platform integrating photoelectrochemical (PEC) and fluorescence (FL) sensors was developed for the sensitive monitoring of heart fatty acid binding protein (h-FABP). First, BiVO4/AgInS2 (BVAIS) composites with excellent photoelectric activity were synthesized as sensing matrices. The BVAIS heterojunction with a well-matched internal energy level structure provided a stable photocurrent. Second, an innovative signal amplification strategy based on octylamine-modified poly(acrylic acid) (OPA)-capped CsPbBr3 (OPCB) nanocrystals (NCs) with excellent catalytic activity and fluorescence property was proposed. On the OPCB nanozyme possessing ascorbate oxidase-like catalytic activity could catalyze the oxidation of ascorbic acid, which achieved quenching of the photocurrent signals by competitively consuming the electron donor. On the other hand, the OPCB NCs that overcame the water stability defect processed good luminescence performance and were able to produce obvious FL signals. Mutually verified dual-response signals effectively enhance the precision of test outcomes and avoid false-positive or false-negative results. Finally, the constructed microfluidic sensing platform realized sensitive detection of h-FABP in the linear range of 0.0001–150 ng/mL (PEC mode) and 0.001–150 ng/mL (FL mode), with detection limits of 36 fg/mL and 0.32 pg/mL, respectively. The present work provided a new perspective for designing an efficient dual-mode sensing strategy to achieve sensitive detection of disease markers.
{"title":"Advanced Dual-Mode Microfluidic Sensing Platform Based on Amphiphilic Polymer-Capped Perovskite Nanozymes Induced Photoelectrochemical Signal Amplification and Fluorescence Emission","authors":"Junqiang Fu, Dongquan Leng, Jingui Chen, Xiang Ren, Yamei Li, Tingting Wu, Qin Wei, Bin Cai","doi":"10.1021/acs.analchem.4c05892","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05892","url":null,"abstract":"A novel dual-mode microfluidic sensing platform integrating photoelectrochemical (PEC) and fluorescence (FL) sensors was developed for the sensitive monitoring of heart fatty acid binding protein (h-FABP). First, BiVO<sub>4</sub>/AgInS<sub>2</sub> (BVAIS) composites with excellent photoelectric activity were synthesized as sensing matrices. The BVAIS heterojunction with a well-matched internal energy level structure provided a stable photocurrent. Second, an innovative signal amplification strategy based on octylamine-modified poly(acrylic acid) (OPA)-capped CsPbBr<sub>3</sub> (OPCB) nanocrystals (NCs) with excellent catalytic activity and fluorescence property was proposed. On the OPCB nanozyme possessing ascorbate oxidase-like catalytic activity could catalyze the oxidation of ascorbic acid, which achieved quenching of the photocurrent signals by competitively consuming the electron donor. On the other hand, the OPCB NCs that overcame the water stability defect processed good luminescence performance and were able to produce obvious FL signals. Mutually verified dual-response signals effectively enhance the precision of test outcomes and avoid false-positive or false-negative results. Finally, the constructed microfluidic sensing platform realized sensitive detection of h-FABP in the linear range of 0.0001–150 ng/mL (PEC mode) and 0.001–150 ng/mL (FL mode), with detection limits of 36 fg/mL and 0.32 pg/mL, respectively. The present work provided a new perspective for designing an efficient dual-mode sensing strategy to achieve sensitive detection of disease markers.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"23 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142937523","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}
Accurate identification of cancer cells under complex physiological environments holds great promise for noninvasive diagnosis and personalized medicine. Herein, we developed dual-aptamer-based DNA logic-gated series lamp probes (DApt-SLP) by coupling a DNA cell-classifier (DCC) with a self-powered signal-amplifier (SSA), enabling rapid and sensitive identification of cancer cells in a blood sample. DCC is endowed with two extended-aptamer based modules for recognizing the two cascade cell membrane receptors and serves as a DNA logic gate to pinpoint a particular and narrow subpopulation of cells from a larger population of similar cells. DCC leverages a dual-receptor co-recognition strategy for enhanced specificity of cell identification by performing the matching operation between aptamer and receptor twice on cell membranes. SSA is a signal converter attached at the end of DCC that changes the cell identification process into detectable signals, as well as a signal amplifier to output amplified signals by using a simple and efficient hybridization chain reaction. Unique from those who are multicomponent systems, DApt-SLP is an all-in-one compact DNA nanodevice, exhibiting an enhanced nuclease-degradation resistance and targeting ability. In vitro feasibility, cell imaging, and flow cytometry analysis showed that the DApt-SLP system successfully operated under buffered solution and physiological environment and precisely differentiated the target cell from large populations of similar cells. Benefiting from its integrated design and single-step cancer cell identification with high sensitivity and accuracy, the DApt-SLP system is a practical tool in personalized medicine and biomedical engineering.
{"title":"Lighting Up Dual-Aptamer-Based DNA Logic-Gated Series Lamp Probes with Specific Membrane Proteins for Sensitive and Accurate Cancer Cell Identification","authors":"Xiaoyu Zhou, Chenghao Yu, Xiaoling Wei, Haiyan Jia, Ling Zheng, Zhifa Shen, Rong Wu, Chang Xue","doi":"10.1021/acs.analchem.4c05505","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05505","url":null,"abstract":"Accurate identification of cancer cells under complex physiological environments holds great promise for noninvasive diagnosis and personalized medicine. Herein, we developed dual-aptamer-based DNA logic-gated series lamp probes (<sup>D</sup>Apt-SLP) by coupling a DNA cell-classifier (DCC) with a self-powered signal-amplifier (SSA), enabling rapid and sensitive identification of cancer cells in a blood sample. DCC is endowed with two extended-aptamer based modules for recognizing the two cascade cell membrane receptors and serves as a DNA logic gate to pinpoint a particular and narrow subpopulation of cells from a larger population of similar cells. DCC leverages a dual-receptor co-recognition strategy for enhanced specificity of cell identification by performing the matching operation between aptamer and receptor twice on cell membranes. SSA is a signal converter attached at the end of DCC that changes the cell identification process into detectable signals, as well as a signal amplifier to output amplified signals by using a simple and efficient hybridization chain reaction. Unique from those who are multicomponent systems, <sup>D</sup>Apt-SLP is an all-in-one compact DNA nanodevice, exhibiting an enhanced nuclease-degradation resistance and targeting ability. In vitro feasibility, cell imaging, and flow cytometry analysis showed that the <sup>D</sup>Apt-SLP system successfully operated under buffered solution and physiological environment and precisely differentiated the target cell from large populations of similar cells. Benefiting from its integrated design and single-step cancer cell identification with high sensitivity and accuracy, the <sup>D</sup>Apt-SLP system is a practical tool in personalized medicine and biomedical engineering.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"6 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940549","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c05585
Teresa Kumblathan, Genievieve C. Borg, Ceilidh Morrissette, Wayne Inuglak Clark, X. Chris Le, Xing-Fang Li
Implementation of Diversity, Equity, Inclusion, and Respect (DEIR) is crucial for supporting students in a culturally safe environment, reducing bias, fostering respect, broadening perspectives, enhancing collaboration, and improving education in science. DEIR with Indigenous reconciliation incorporates Indigenous-based DEIR initiatives as a response to the Truth and Reconciliation Commission (TRC) in Canada to acknowledge the intergenerational trauma and mistrust toward colonial institutions such as universities. Universities can advance reconciliation by incorporating DEIR with Indigenous reconciliation into everyday practices. Indigenous students are significantly less likely to attain degrees in science, technology, engineering, and mathematics (STEM). The lack of Indigenous representation in STEM significantly hinders the inclusion of Indigenous perspectives in scientific processes and decision-making. A collaborative effort is essential to improve Indigenous student recruitment in science programs. Scientists need to educate themselves on the colonial legacies of Indian Residential Schools (IRSs) and Indian Hospitals (IHs), as well as on the biases and barriers that Indigenous students face. While this paper focuses on several Canadian examples, it highlights challenges and traumas that are similarly faced by Indigenous students worldwide. Therefore, supervisors and research groups should actively participate in training sessions and develop strategies aimed at preventing discrimination and fostering inclusivity. This paper highlights the importance of DEIR with Indigenous reconciliation in university science programs and addresses issues of recruiting Indigenous students. The “Calls to Action” outlined in this paper will help scientists and educators understand barriers faced by Indigenous students, advocate for lifelong learning and social stewardship, and foster a more inclusive scientific environment.
{"title":"Advancing Equity and Empowering Science Students from Indigenous Communities","authors":"Teresa Kumblathan, Genievieve C. Borg, Ceilidh Morrissette, Wayne Inuglak Clark, X. Chris Le, Xing-Fang Li","doi":"10.1021/acs.analchem.4c05585","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05585","url":null,"abstract":"Implementation of Diversity, Equity, Inclusion, and Respect (DEIR) is crucial for supporting students in a culturally safe environment, reducing bias, fostering respect, broadening perspectives, enhancing collaboration, and improving education in science. DEIR with Indigenous reconciliation incorporates Indigenous-based DEIR initiatives as a response to the Truth and Reconciliation Commission (TRC) in Canada to acknowledge the intergenerational trauma and mistrust toward colonial institutions such as universities. Universities can advance reconciliation by incorporating DEIR with Indigenous reconciliation into everyday practices. Indigenous students are significantly less likely to attain degrees in science, technology, engineering, and mathematics (STEM). The lack of Indigenous representation in STEM significantly hinders the inclusion of Indigenous perspectives in scientific processes and decision-making. A collaborative effort is essential to improve Indigenous student recruitment in science programs. Scientists need to educate themselves on the colonial legacies of Indian Residential Schools (IRSs) and Indian Hospitals (IHs), as well as on the biases and barriers that Indigenous students face. While this paper focuses on several Canadian examples, it highlights challenges and traumas that are similarly faced by Indigenous students worldwide. Therefore, supervisors and research groups should actively participate in training sessions and develop strategies aimed at preventing discrimination and fostering inclusivity. This paper highlights the importance of DEIR with Indigenous reconciliation in university science programs and addresses issues of recruiting Indigenous students. The “Calls to Action” outlined in this paper will help scientists and educators understand barriers faced by Indigenous students, advocate for lifelong learning and social stewardship, and foster a more inclusive scientific environment.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"15 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142937519","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c05665
Yea-Rin Lee, Ibrahim Kaya, Elin Wik, Sooraj Baijnath, Henrik Lodén, Anna Nilsson, Xiaoqun Zhang, Dag Sehlin, Stina Syvänen, Per Svenningsson, Per E. Andrén
Multiomics analysis of single tissue sections using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) provides comprehensive molecular insights. However, optimizing tissue sample preparation for MALDI-MSI to achieve high sensitivity and reproducibility for various biomolecules, such as lipids, N-glycans, and tryptic peptides, presents a significant challenge. This study introduces a robust and reproducible protocol for the comprehensive sequential analysis of the latter molecules using MALDI-MSI in fresh-frozen rodent brain tissue samples. The optimization process involved testing multiple organic solvents, which identified serial washing in ice-cold methanol, followed by chloroform as optimal for N-glycan analysis. Integrating this optimized protocol into MALDI-MSI workflows enabled comprehensive sequential analysis of lipids (in dual polarity mode), N-glycans, and tryptic peptides within the same tissue sections, enhancing both the efficiency and reliability. Validation across diverse rodent brain tissue samples confirmed the protocol’s robustness and versatility. The optimized methodology was subsequently applied to a transgenic Alzheimer’s disease (AD) mouse model (tgArcSwe) as a proof of concept. In the AD model, significant molecular alterations were observed in various sphingolipid and glycerophospholipid species, as well as in biantennary and GlcNAc-bisecting N-glycans, particularly in the cerebral cortex. These region-specific alterations are potentially associated with amyloid-beta (Aβ) plaque accumulation, which may contribute to cognitive and memory impairments. The proposed standardized methodology represents a significant advancement in neurobiological research, providing valuable insights into disease mechanisms and laying the foundation for potential preclinical applications. It could aid the development of diagnostic biomarkers and targeted therapies for AD and other neurodegenerative diseases, such as Parkinson’s disease.
{"title":"Comprehensive Approach for Sequential MALDI-MSI Analysis of Lipids, N-Glycans, and Peptides in Fresh-Frozen Rodent Brain Tissues","authors":"Yea-Rin Lee, Ibrahim Kaya, Elin Wik, Sooraj Baijnath, Henrik Lodén, Anna Nilsson, Xiaoqun Zhang, Dag Sehlin, Stina Syvänen, Per Svenningsson, Per E. Andrén","doi":"10.1021/acs.analchem.4c05665","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05665","url":null,"abstract":"Multiomics analysis of single tissue sections using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) provides comprehensive molecular insights. However, optimizing tissue sample preparation for MALDI-MSI to achieve high sensitivity and reproducibility for various biomolecules, such as lipids, <i>N</i>-glycans, and tryptic peptides, presents a significant challenge. This study introduces a robust and reproducible protocol for the comprehensive sequential analysis of the latter molecules using MALDI-MSI in fresh-frozen rodent brain tissue samples. The optimization process involved testing multiple organic solvents, which identified serial washing in ice-cold methanol, followed by chloroform as optimal for <i>N</i>-glycan analysis. Integrating this optimized protocol into MALDI-MSI workflows enabled comprehensive sequential analysis of lipids (in dual polarity mode), <i>N</i>-glycans, and tryptic peptides within the same tissue sections, enhancing both the efficiency and reliability. Validation across diverse rodent brain tissue samples confirmed the protocol’s robustness and versatility. The optimized methodology was subsequently applied to a transgenic Alzheimer’s disease (AD) mouse model (tgArcSwe) as a proof of concept. In the AD model, significant molecular alterations were observed in various sphingolipid and glycerophospholipid species, as well as in biantennary and GlcNAc-bisecting <i>N</i>-glycans, particularly in the cerebral cortex. These region-specific alterations are potentially associated with amyloid-beta (Aβ) plaque accumulation, which may contribute to cognitive and memory impairments. The proposed standardized methodology represents a significant advancement in neurobiological research, providing valuable insights into disease mechanisms and laying the foundation for potential preclinical applications. It could aid the development of diagnostic biomarkers and targeted therapies for AD and other neurodegenerative diseases, such as Parkinson’s disease.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"28 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142937527","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c06318
Libing Ke, Chenji Dai, Yaoyao Xu, Yuyang Zhou
Pneumonia is a prevalent acute respiratory infection and a major cause of mortality and hospitalization, and the urgent demand for a rapid, direct, and highly accurate diagnostic method capable of detecting both Streptococcus pneumoniae (S. pneumoniae) and Klebsiella pneumoniae (K. pneumoniae) arises from their prominent roles as the primary pathogens responsible for pneumonia. Herein, two luminescent iridium complexes with nonoverlapping photoluminescence spectra, iridium(III)-bis [4,6-(difluorophenyl)-pyridinato-N,C2′] picolinate (abbreviated as Ir–B) and bis (2-(3,5- dimethylphenyl) quinoline-C2,N′) (acetylacetonato) iridium(III)) (abbreviated as Ir-R), were unprecedently proposed to construct a novel wavelength-resolved magnetic multiplex biosensor for simultaneous detection of S. pneumoniae and K. pneumoniae based on catalytic hairpin assembly (CHA) signal amplification strategy combined with dye-doped silica nanoparticles. Notably, the proposed wavelength-resolved multiplex biosensor not only exhibits a broad linear range from 50 pM to 10 nM but also demonstrates excellent recovery rates for S. pneumoniae (96.1–99.3%) and K. pneumoniae (94.8–101.5%) in real clinical samples, with corresponding relative standard deviation (RSD) values ranging from 2.57 to 3.15% for S. pneumoniae and 1.45 to 3.17% for K. pneumoniae. These favorable experimental outcomes undoubtedly offer a promising approach for the simultaneous detection of multiple pathogens in the future.
{"title":"Wavelength-Resolved Magnetic Multiplex Biosensor for Simultaneous and Ultrasensitive Detection of Pneumonia Pathogens via Catalytic Hairpin Assembly Strategy with Luminescent Iridium Complexes","authors":"Libing Ke, Chenji Dai, Yaoyao Xu, Yuyang Zhou","doi":"10.1021/acs.analchem.4c06318","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c06318","url":null,"abstract":"Pneumonia is a prevalent acute respiratory infection and a major cause of mortality and hospitalization, and the urgent demand for a rapid, direct, and highly accurate diagnostic method capable of detecting both <i>Streptococcus pneumoniae</i> (<i>S. pneumoniae</i>) and <i>Klebsiella pneumoniae</i> (<i>K. pneumoniae</i>) arises from their prominent roles as the primary pathogens responsible for pneumonia. Herein, two luminescent iridium complexes with nonoverlapping photoluminescence spectra, iridium(III)-bis [4,6-(difluorophenyl)-pyridinato-N,C<sup>2</sup>′] picolinate (abbreviated as Ir–B) and bis (2-(3,5- dimethylphenyl) quinoline-C2,N′) (acetylacetonato) iridium(III)) (abbreviated as Ir-R), were unprecedently proposed to construct a novel wavelength-resolved magnetic multiplex biosensor for simultaneous detection of <i>S. pneumoniae</i> and <i>K. pneumoniae</i> based on catalytic hairpin assembly (CHA) signal amplification strategy combined with dye-doped silica nanoparticles. Notably, the proposed wavelength-resolved multiplex biosensor not only exhibits a broad linear range from 50 pM to 10 nM but also demonstrates excellent recovery rates for <i>S. pneumoniae</i> (96.1–99.3%) and <i>K. pneumoniae</i> (94.8–101.5%) in real clinical samples, with corresponding relative standard deviation (RSD) values ranging from 2.57 to 3.15% for <i>S. pneumoniae</i> and 1.45 to 3.17% for <i>K. pneumoniae</i>. These favorable experimental outcomes undoubtedly offer a promising approach for the simultaneous detection of multiple pathogens in the future.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"39 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142940563","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c05319
Iakovos Saridakis, Kish R. Adoni, Thomas Leischner, Bogdan R. Brutiu, Saad Shaaban, Giammarco Ferrari, Konstantinos Thalassinos, Nuno Maulide
Chemical cross-linking/mass spectrometry (XL-MS) has emerged as a complementary tool for mapping interaction sites within protein networks as well as gaining moderate-resolution native structural insight with minimal interference. XL-MS technology mostly relies on chemoselective reactions (cross-linking) between protein residues and a linker. DSSO represents a versatile cross-linker for protein structure investigation and in-cell XL-MS. However, our assessment of its shelf life and batch purity revealed decomposition of DSSO in anhydrous solution via a retro-Michael reaction, which may reduce the active ingredient down to below 90%. To mitigate the occurrence of this degradative mechanism, we report the rational design and synthesis of DSSO-carbamate, which contains an inserted nitrogen atom in the DSSO backbone structure. This modification to DSSO yielded remarkably favorable stability against such decomposition, which translated to higher cross-link and monolink recovery when performing XL-MS on monomeric flexible proteins. Recently, XL-MS has been leveraged against AlphaFold2 and other protein structure prediction algorithms for improved prediction of flexible monomeric multiconformational proteins. To this end, we demonstrate that our novel cross-linker, termed DSSO-carbamate, generated more accurate protein structure predictions when combined with AlphaFold2, on account of its increased recovery of cross-links and monolinks, compared to DSSO. As such, DSSO-carbamate represents a useful addition to the XL-MS community, particularly for protein structure prediction.
{"title":"Rational Modification of a Cross-Linker for Improved Flexible Protein Structure Modeling","authors":"Iakovos Saridakis, Kish R. Adoni, Thomas Leischner, Bogdan R. Brutiu, Saad Shaaban, Giammarco Ferrari, Konstantinos Thalassinos, Nuno Maulide","doi":"10.1021/acs.analchem.4c05319","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c05319","url":null,"abstract":"Chemical cross-linking/mass spectrometry (XL-MS) has emerged as a complementary tool for mapping interaction sites within protein networks as well as gaining moderate-resolution native structural insight with minimal interference. XL-MS technology mostly relies on chemoselective reactions (cross-linking) between protein residues and a linker. DSSO represents a versatile cross-linker for protein structure investigation and in-cell XL-MS. However, our assessment of its shelf life and batch purity revealed decomposition of DSSO in anhydrous solution via a retro-Michael reaction, which may reduce the active ingredient down to below 90%. To mitigate the occurrence of this degradative mechanism, we report the rational design and synthesis of DSSO-carbamate, which contains an inserted nitrogen atom in the DSSO backbone structure. This modification to DSSO yielded remarkably favorable stability against such decomposition, which translated to higher cross-link and monolink recovery when performing XL-MS on monomeric flexible proteins. Recently, XL-MS has been leveraged against AlphaFold2 and other protein structure prediction algorithms for improved prediction of flexible monomeric multiconformational proteins. To this end, we demonstrate that our novel cross-linker, termed DSSO-carbamate, generated more accurate protein structure predictions when combined with AlphaFold2, on account of its increased recovery of cross-links and monolinks, compared to DSSO. As such, DSSO-carbamate represents a useful addition to the XL-MS community, particularly for protein structure prediction.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"55 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142937419","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 : 2025-01-09DOI: 10.1021/acs.analchem.4c04065
So Jin Hong, Eunsol Jeon, Min Jung Kim, Min Hee Lee
Reductase expression is a potential indicator of cellular pathology. Single-detection systems for reductases have been developed, however, the development of dual-detection systems remain largely unexplored. We rationally designed a dual-lock fluorescent probe that exhibited a high signal-to-noise ratio with a fluorescence Off-On response exclusively for the simultaneous activity of two reductases, NTR and hNQO1, which are overexpressed in cancer hypoxia. The system comprised a naphthalimide fluorophore with dual-lock control mediated by PET and ICT, a trimethyl-locked quinone group sensitive to hNQO1, and a nitrobenzyl carbamate group sensitive to NTR. This study employed a hypoxia model in HeLa cells to demonstrate that our developed dual-lock system detected hypoxia more effectively than single-detection systems. Moreover, it enabled noninvasive real-time monitoring of hypoxia in zebrafish embryos. Consequently, the dual-lock fluorescent probe, which strategically provides a fluorescence response only when both NTR and NQO1 are active, offers a novel diagnostic platform for both in vitro and in vivo applications, effectively detecting hypoxia and monitoring various pathological states.
{"title":"Dual-Lock System for High Sensitivity and Selectivity in Redox Enzyme Activation and Imaging","authors":"So Jin Hong, Eunsol Jeon, Min Jung Kim, Min Hee Lee","doi":"10.1021/acs.analchem.4c04065","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04065","url":null,"abstract":"Reductase expression is a potential indicator of cellular pathology. Single-detection systems for reductases have been developed, however, the development of dual-detection systems remain largely unexplored. We rationally designed a dual-lock fluorescent probe that exhibited a high signal-to-noise ratio with a fluorescence Off-On response exclusively for the simultaneous activity of two reductases, NTR and hNQO1, which are overexpressed in cancer hypoxia. The system comprised a naphthalimide fluorophore with dual-lock control mediated by PET and ICT, a trimethyl-locked quinone group sensitive to hNQO1, and a nitrobenzyl carbamate group sensitive to NTR. This study employed a hypoxia model in HeLa cells to demonstrate that our developed dual-lock system detected hypoxia more effectively than single-detection systems. Moreover, it enabled noninvasive real-time monitoring of hypoxia in zebrafish embryos. Consequently, the dual-lock fluorescent probe, which strategically provides a fluorescence response only when both NTR and NQO1 are active, offers a novel diagnostic platform for both in vitro and in vivo applications, effectively detecting hypoxia and monitoring various pathological states.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"83 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142937522","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}
An in situ monitoring reaction can better obtain the variations during the progression of the photocatalytic reaction. However, the complexity of the apparatus and the limited applicability of substances are the common challenges faced by most in situ monitoring methods. Here, we invented an in situ infrared optical fiber sensor to monitor the reactants and products during photocatalytic reaction. The sensor, which has four tapered regions, demonstrates the best sensitivity of 0.71 au/vol %, 70 times higher than that of the fiber sensor without a tapered region. Then, this sensor was successfully used to in situ monitor the photocatalytic reaction between benzaldehyde and ethanol under the UV light and TiO2. The calibration plots of the reactants and products were established by sensing a series of designed concentration solutions. Based on the calibration plots, the real-time concentrations of four substances could be derived by converting the absorbance values, and the concentration changes of the reactants and products followed the first order kinetic mode. The equilibrium concentrations of reactants and products could be obtained from the fitting curves. With the increase in the UV light intensity, this sensor could detect a gradual increase in the rate of this photocatalytic reaction. The results show that this in situ infrared fiber sensor can monitor the progression of the photocatalytic reaction in real time, which will be helpful for unveiling the photocatalytic mechanism.
{"title":"In Situ Infared Optical Fiber Sensor Monitoring Reactants and Products Changes during Photocatalytic Reaction","authors":"Zeyan Wu, Yongkun Zhao, Tianxiang You, Zongkui Kou, Yantao Xu, Mengling Xia, Haitao Guo, Xianghua Zhang, Yinsheng Xu","doi":"10.1021/acs.analchem.4c04704","DOIUrl":"https://doi.org/10.1021/acs.analchem.4c04704","url":null,"abstract":"An in situ monitoring reaction can better obtain the variations during the progression of the photocatalytic reaction. However, the complexity of the apparatus and the limited applicability of substances are the common challenges faced by most in situ monitoring methods. Here, we invented an in situ infrared optical fiber sensor to monitor the reactants and products during photocatalytic reaction. The sensor, which has four tapered regions, demonstrates the best sensitivity of 0.71 au/vol %, 70 times higher than that of the fiber sensor without a tapered region. Then, this sensor was successfully used to in situ monitor the photocatalytic reaction between benzaldehyde and ethanol under the UV light and TiO<sub>2</sub>. The calibration plots of the reactants and products were established by sensing a series of designed concentration solutions. Based on the calibration plots, the real-time concentrations of four substances could be derived by converting the absorbance values, and the concentration changes of the reactants and products followed the first order kinetic mode. The equilibrium concentrations of reactants and products could be obtained from the fitting curves. With the increase in the UV light intensity, this sensor could detect a gradual increase in the rate of this photocatalytic reaction. The results show that this in situ infrared fiber sensor can monitor the progression of the photocatalytic reaction in real time, which will be helpful for unveiling the photocatalytic mechanism.","PeriodicalId":27,"journal":{"name":"Analytical Chemistry","volume":"35 1","pages":""},"PeriodicalIF":7.4,"publicationDate":"2025-01-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142937520","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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