Analytical and Bioanalytical Chemistry最新文献

{"title":"A hemicyanine-based dual-responsive fluorescent sensor for the detection of lithium and cyanide ions: application in living cells.","authors":"Ziya Aydin, Mukaddes Keskinates, Esra Armagan, Bahar Yilmaz Altinok, Mevlut Bayrakci","doi":"10.1007/s00216-025-05852-w","DOIUrl":"10.1007/s00216-025-05852-w","url":null,"abstract":"<p><p>A hemicyanine-based colorimetric and fluorometric sensor, 2-(2-(2,3,5,6,8,9-hexahydrobenzo[b][1,4,7,10]tetraoxacyclododecin-12-yl)vinyl)-3,3-dimethyl-1-propyl-3H-indol-1-ium iodide (MH-5), was developed and synthesized to detect Li⁺ and CN^- ions in DMSO-PBS buffer solution (10 mM, pH 7.25, v/v 1:9). MH-5 displayed a rapid and highly selective colorimetric response to both Li⁺ and CN^-, indicated by a distinct color change from pink to pale pink in the presence of Li⁺ and to colorless upon CN^- detection, without interference from other cations or anions. The interaction mechanisms of MH-5 with Li⁺ and CN^- ions were investigated using various analytical techniques, including ¹H NMR, ESI-MS, FT-IR spectroscopy, and Job's plot analysis. These studies suggest that CN^- is detected through nucleophilic addition to the indolium moiety of MH-5, while Li⁺ detection occurs via coordination with oxygen atoms in the crown ether structure. The fluorescence-based detection limits for Li⁺ and CN^- were determined to be 0.150 µM and 0.154 µM, respectively. Additionally, MH-5 was evaluated in living cells, demonstrating effective cell penetration and reliable detection of Li⁺ and CN^- ions for potential bio-imaging applications.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3127-3139"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103329/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143778714","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pyrolysis-GC/MS differentiates polyesters and detects additives for improved monitoring of textile labeling accuracy and plastic pollution.","authors":"Josh Forakis, Jennifer Lynch","doi":"10.1007/s00216-025-05851-x","DOIUrl":"10.1007/s00216-025-05851-x","url":null,"abstract":"<p><p>Polyesters comprise the greatest proportion of textile fibers and are found in various everyday goods; hence, polyester fibers are a significant source of microplastic pollution and textile waste. The specific chemical composition of commercial polyester fibers is often proprietary and mostly assumed to be poly(ethylene terephthalate) (PET). Polyester is a class of polymers that include poly(butylene terephthalate) (PBT), poly(cyclohexylenedimethylene terephthalate) (PCT), and poly(ethylene naphthalate) (PEN), as well as biodegradable polymers. Our study aims to clarify whether household polyester products are primarily PET, are labeled accurately, or contain phthalate additives by applying double-shot pyrolysis-gas chromatography/mass spectroscopy (Py-GC/MS). We analyzed four scientific-grade polyester reference standards, 52 manufacturer-grade polyester fibers or pellets, and 229 samples from 193 consumer polyester products. From the pyrograms, samples were predominantly identified as PET (87.4%, 95% CI [93.5-81.3%]), but five samples were identified as a different polyester, nine as non-polyester polymers, and 23 as a blend of PET with another polymer. From the thermal desorption chromatograms, diethyl phthalate was the most frequently detected phthalate, found in 23.3% (95% CI [17.3-29.3%]) of the consumer products, including children's toys. Double-shot py-GC/MS advantageously results in these empirical data that (1) counter the assumption that products labeled polyester are always PET, (2) emphasize the importance of creating spectral libraries with well-characterized materials for accurate polymer identification of unknown plastic particles, and (3) demonstrate that phthalates are common additives in household products.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3113-3126"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103379/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Direct transfer of multicellular tumor spheroids grown in agarose microarrays for high-throughput mass spectrometry imaging analysis.","authors":"Yuan Liu, Jillian Johnson, Hua Zhang, Penghsuan Huang, Lingjun Li","doi":"10.1007/s00216-025-05843-x","DOIUrl":"10.1007/s00216-025-05843-x","url":null,"abstract":"<p><p>Multicellular tumor spheroids (MCTSs) play an important role in biological studies and cancer research. There is an emerging research interest in molecular profiling and drug distribution of MCTSs by leveraging the superior sensitivity and molecular specificity of mass spectrometry imaging (MSI). Current methods for sample preparation of MCTSs can suffer from low throughput, as MCTSs are typically individually transferred from cell culture into an MSI embedding media and sectioned individually, or sometimes, a few spheroids are placed in a small block of embedding media in preparation for MSI. Here, we developed a method to minimize the sample preparation steps needed to create high-throughput MCTS frozen sections for MSI. Agarose-based microarrays created from Microtissues^® molds were used during MCTS culturing, after which the entire MCTS agarose microarray was taken out of the cell culture well and then directly embedded in 5% gelatin, without the need for a transfer step for each individual MCTS into the embedding media. This method enables rapid profiling of up to 81 MCTSs for larger MCTSs (500-800 µm) or up to 256 MCTSs for smaller MCTSs (200-300 µm) in a single section, remarkably improving the throughput possible for MSI MCTS workflows. Notably, sectioning MCTSs together in the agarose microarray also improves MCTS visualization during sectioning, such that staining each MCTS section to ensure the presence of the MCTSs within the embedding media is not necessary during the sectioning process. The method described here provides a more direct, convenient strategy to achieve high-throughput sections. MSI MCTS sectioning throughput is an important advancement for both pharmaceutical testing of MCTS; the direct transfer 3D cell cultures grown within cell culture-compatible polymer scaffolding are also critical for expanding MSI for the characterization of microfluidic and complex in vitro models, where agarose is readily utilized as a non-adhesive 3D cell culture scaffold.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3021-3031"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103265/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143741940","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Kinetic and analytical characterization of a new tropinone oxidase enzyme and its application to the simultaneous determination of the tropane alkaloids atropine and scopolamine.","authors":"Mario Domínguez, Susana de Marcos, Javier Galbán","doi":"10.1007/s00216-025-05856-6","DOIUrl":"10.1007/s00216-025-05856-6","url":null,"abstract":"<p><p>A spectrophotometric enzymatic method for the determination of atropine (Atp) and scopolamine (Scp), two tropane alkaloids (TAs), has been developed. The method is based on a previous basic hydrolysis to tropine (Trp) and scopine (Sci) respectively, and a subsequent enzymatic oxidation catalyzed by a tropinone reductase 1 (TRase) using NAD as oxidant; the absorbance of NADH (340 nm) is monitored during the reaction. First, the enzyme kinetics of both substrates are studied; it is concluded that both TAs follow a compulsory order ternary complex mechanism and the Michalis-Menten constant is calculated. Then, an enzymatic method was optimized for Atp, allowing the determination of this substrate in the range from 1.1·10^-5 M to 3.0·10^-4 M (LoD = 3.5·10^-6 M); it was applied to the determination of Atp in a spiked chia sample (96 ± 6% recovery). Interestingly, the equilibrium constant of the reaction decreased with temperature and increased with enzyme concentration; both effects were satisfactorily explained. A similar analytical study was carried out with Scp (linear range from 1.2·10^-5 M to 3.0·10^-4 M, LoD = 3.6·10^-6 M); the method was also applied to Scp in a spiked chia sample (94 ± 2% recovery). Finally, since the kinetics of Scp is slower than that of Atp, the simultaneous quantitative determination of both compounds was successfully developed by measuring the absorbance at two reaction times (70 s and 300 s). This method was applied to the simultaneous determination of both TAs first in a synthetic sample and later in a spiked chia sample, with recoveries around 98% for both compounds. Although the sensitivity of the method is lower than that of the immunoassays for Atp, it has advantages such as the simultaneous determination of Atp and Scp, and even the possible determination of Trp (another TA).</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3169-3176"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103354/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957494","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Study of sampling phases for body odor sampling prior to analysis by TD-GC×GC/ToFMS.","authors":"Elsa Boudard, Lisa Fisson, Nabil Moumane, José Dugay, Jérôme Vial, Didier Thiébaut","doi":"10.1007/s00216-025-05857-5","DOIUrl":"10.1007/s00216-025-05857-5","url":null,"abstract":"<p><p>Body odor consists of a complex matrix of volatile organic compounds (VOCs), which has garnered increasing interest in fields like medicine for its potential in disease diagnosis. However, the field of body odor analysis is advancing slowly, partly due to a lack of standardized methodologies. Although gas chromatography-mass spectrometry (GC-MS) is widely used for VOC analysis, there is a broad range of sampling and extraction methods, leading to different or even sometimes contradictory results. To move toward standardized procedures, this study compares five sampling phases for direct body odor sampling in terms of analytical cleanliness and VOC trapping/release efficiency: gauze, glass beads, PowerSorb^®, Getxent^® microtubes, and passive sampling pillows (PSP). Thermodesorption was employed to simplify the protocol and minimize contamination or sample loss, which often occurs during multistep processes. Given the matrix's complexity and the need to detect trace-level compounds, comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC×GC/ToFMS) was used to achieve high sensitivity and peak capacity. PSP and PowerSorb^® demonstrated the best performance, with mean recovery yields of 95% and 71%, respectively, and 22% and 10% variability, ensuring good repeatability. These findings, initially obtained under simulated conditions with a synthetic mixture, were validated with real body odor samples, with an optimal sampling duration estimated between 30 min and 1 h. This study not only highlights these effective sampling solutions but also emphasizes the risks associated with using sorbent phases that lack adequate analytical cleanliness (i.e., clean blank) such as gauze.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3177-3190"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103381/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143957077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering DNA circuit powered by entropy integrated into robust and elegant photoelectrochemical and photothermal dual-mode biosensing.","authors":"Jing Li, Bingtao Hu, Yanxin Zhang, Qin Xu, Hongbo Li","doi":"10.1007/s00216-025-05848-6","DOIUrl":"10.1007/s00216-025-05848-6","url":null,"abstract":"<p><p>Dual-signal mode sensors that can self-validate detection results have attracted considerable interest; however, creating those with superior overall performance still presents significant challenges. Herein, we develop a unique photoelectrochemical (PEC) and photothermal (PT) dual-mode biosensor targeting microRNA-221 (miRNA-221), built on an innovative entropy-driven DNA circuit (EDC). The zinc oxide nanorods (ZnO NRs) serve as PEC beacons, while copper sulfide nanoparticles (CuS NPs) function as photocurrent inhibitors and PT beacons, both biofunctionalized with DNAs before being assembled through partial base pairing. When target miRNA-221 is present, the EDC activates and releases output DNAs that open partially hybridized strands anchored to ZnO NRs via competitive assembly. This process liberates CuS-DNA1 and restores the suppressed photocurrent. The results demonstrate linear relationships between photocurrent/temperature increment and the logarithm of target concentration across ranges of 1.0 fmol L^-1-50.0 pmol L^-1 (limit of detection (LOD): 0.35 fmol L^-1) and 5.0×10² fmol L^-1-5.0 nmol L^-1 (LOD: 1.22×10² fmol L^-1), respectively. Compared to conventional EDCs, our optimally designed EDC not only doubles the output DNA yield but also significantly enhances sensor sensitivity. Additionally, the target-triggered EDC amplification strategy effectively minimizes reversibility in each reaction step, preserves base sequence integrity, boosts efficiency, and demonstrates strong thermal stability and selectivity, thereby increasing the specificity of the dual-mode biosensor. Furthermore, ZnO NR photoelectric beacons fabricated via electrodeposition greatly improve the stability and controllability of the photoelectrode while avoiding lengthy modification processes. Overall, this thoughtfully engineered dual-mode biosensor offers numerous advantages, including a wide linear range, excellent stability, high reproducibility, and user-friendly operation. Specifically, this signal-on type dual-signal output biosensor enables self-confirmation of detection results, significantly enhancing both accuracy and reliability.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3079-3089"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143708093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of microprefilter for chromatography column in HbA1c assays.","authors":"Zhiyan Li, Hongying Wang, Le Chang, Cunling Yan","doi":"10.1007/s00216-025-05844-w","DOIUrl":"10.1007/s00216-025-05844-w","url":null,"abstract":"<p><p>The complexity of blood and its matrix in high-performance liquid chromatography (HPLC) glycated hemoglobin (HbA1c) assays is closely related to column performance. However, the available prefilters with a single structure or surface membrane materials are not ideal for column protection, and coupled with the complexity of blood samples, leads to rapid degradation of column performance. Therefore, we have developed a new microprefilter with a three-stage filtration design and depth filter material to protect the column. All filter materials used in the preparation of microprefilters were characterized, screened, and optimized, and then manufactured on the basis of optimized filter materials, which are depth filter material microprefilters. Based on the material and structural design, microprefilters were capable of filtering particulate matter from test samples on a step-by-step basis to avoid the plugging effect that occurs when all sizes of substances are gathered together. Moreover, all newly developed microprefilters can be tested more times, up to 600 times. Microprefilters with small-pore-size final filtration membranes of polyethersulfone, hydrophilic polytetrafluoroethylene, and mixed cellulose showed excellent column protection in terms of column efficiency, HbA1c retention time, number of column tests, and column backpressure, and prolonged column lifetime by as much as 20-30% compared with microprefilters with large-pore-size final membranes. Our study provides valuable depth filter material microprefilters with multistage filtration for chromatography columns, and showed excellent column protection and prolonged column lifetime. Meanwhile, microprefilters can be tested more times. The newly developed microprefilters with a small-pore-size final membrane are the optimal choice for column protection of the HbA1c assay.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3033-3045"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143735579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantitative ¹H NMR optimization for high-throughput metabolite analysis in industrial bioprocess monitoring.","authors":"Yingting Shi, Yuxiang Wan, Yiru Wang, Kerui Fang, Jiayu Yang, Yuting Lu, Xinyuan Xie, Jianyang Pan, Dong Gao, Haibin Wang, Haibin Qu","doi":"10.1007/s00216-025-05845-9","DOIUrl":"10.1007/s00216-025-05845-9","url":null,"abstract":"<p><p>Quantitative ¹H NMR (¹H qNMR) is an ideal tool for bioprocess monitoring because it can comprehensively detect and quantify diverse metabolites that significantly influence bioprocess performance. However, the long experiment time associated with the ¹H qNMR, due to the long longitudinal relaxation time (T1) of some metabolites, does not meet the requirements for high-throughput analysis. We developed a high-throughput ¹H qNMR method for bioprocess analysis using a short relaxation delay (D1) to reduce analytical time and a correction factor (k) to compensate for incomplete relaxation. A total of 27 metabolites were quantified using spectral deconvolution via a peak fitting algorithm and MCR-ALS. Methodological validation results indicated that the precision and accuracy of the developed qNMR method were consistently high across different D1 values, with LOQs ranging from 0.008 to 0.13 mM and LODs ranging from 0.024 to 0.38 mM. Notably, a longer D1 value generally resulted in lower LODs and LOQs for most metabolites. A D1 value of 4 s was optimal for balancing analysis time and performance. The method is broadly applicable for bioprocess monitoring and control, offering valuable guidance for optimizing CHO cell culture processes and improving yield.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"3047-3059"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750566","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Total organic carbon (TOC): a simple tool for assessing micro(nano)plastics and nanocellulose recovery during size-based fractionation.","authors":"Guillaume Bucher, Gabriella F Schirinzi, Chiara Verra, Hind El Hadri, Otmar Geiss, Douglas Gilliland","doi":"10.1007/s00216-025-05812-4","DOIUrl":"10.1007/s00216-025-05812-4","url":null,"abstract":"<p><p>The assessment of analyte recovery during sample preparation is a critical quality control parameter in method development. While elemental mass spectrometry techniques, such as ICP-MS, are very effective for assessing the recovery of particulate materials containing metallic elements, there is no equivalent applicable to metal-free carbon or CHNO-based particulate polymer materials. Vibrational spectro-microscopy or thermo-analytical techniques can be used to quantify polymer-based micro- and nanoparticles, but are typically expensive and time-consuming techniques that require higher levels of expertise. This study investigated the potential of a liquid-based total organic carbon (TOC) analyzer as a simple, cost-effective, and universal method for determining the recovery of polymer-based particulate micro- and nanomaterials following filtration, centrifugation, and asymmetric flow field flow fractionation (AF4) processes. A good correlation between solid contents and TOC analysis was demonstrated for standard polystyrene (PS) particle suspensions of various sizes, ranging from 50 nm to 90 μm (79.2 to 113.6% recovery), and other types of synthetic and natural polymeric particle suspensions, including polyethylene (PE), polypropylene (PP), polyvinyl chloride (PVC), polyethylene terephthalate (PET), and cellulose (86.2 to 126.2% recovery). Liquid-based TOC was then successfully applied to estimate particle recovery after various preparatory and fractionation steps, including the determination of filtration recoveries for nanocellulose suspensions (99.0 to 101.4% recovery) and PS micro- and nanoparticles spiked into environmental lake and river freshwater samples (70 to 96% recovery). The combination of TOC and single particle extinction and scattering (SPES) measurements allowed the tracking and quantification of three different populations of PS particles in a mixture (200, 500, and 1000 nm) during successive centrifugation steps (113.8 ± 13.9% cumulative recovery). Finally, this study demonstrated the suitability of TOC for determining both the absolute and relative recoveries of polymer-based particulate materials after AF4 fractionation in line with ISO standards. Liquid-based TOC proved to be a valuable tool for directly tracking, quantifying, and evaluating the recovery of polymer-based micro- and nanoparticles in model and environmental water samples before and after routine size-based fractionation steps.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"2983-2996"},"PeriodicalIF":3.8,"publicationDate":"2025-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103321/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144092300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid and highly sensitive immunoassay using an ultra-thin immuno-wall microfluidic device with a sequential fluorescence signal increment method.","authors":"Xiang Zhou, Toshihiro Kasama, Ryo Miyake","doi":"10.1007/s00216-025-05916-x","DOIUrl":"https://doi.org/10.1007/s00216-025-05916-x","url":null,"abstract":"<p><p>We present a rapid and highly sensitive immunoassay platform based on an ultra-thin immuno-wall microfluidic device with an easy-to-perform sequential fluorescence signal increment method. The ultra-thin immuno-wall was fabricated using a special type of water-soluble photopolymer mixed with streptavidin via photolithography. During photolithography, the photopolymer formed a three-dimensional cross-linked structure, and streptavidin was immobilized in the cross-linked structure based on the click chemistry reaction. The immobilized streptavidin was used to immobilize biotin-conjugated antibodies on the cross-linked structure to capture biomarkers, forming immune complexes on the surface, known as an \"immuno-wall.\" A sequential fluorescence signal increment method utilizes two different fluorescence-labeled antibodies with high affinity that were incubated several cycles in the immuno-wall to enhance the fluorescence signal. Moreover, an ultra-thin immuno-wall was developed to reduce the nonspecific binding and increase the signal-to-noise ratio. To evaluate the performance of this immunoassay platform, the spike protein from the SARS-CoV-2 virus was selected as the target biomarker. This immunoassay platform exhibited a limit of detection of 0.01 ng/mL, and the detection time was 30 min, which is comparable to rapid antigen tests. This immunoassay platform demonstrates significant potential for early-phase disease diagnosis.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144155343","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}