Springer最新文献

{"title":"Designing the homogeneous competitive bioluminescence-based assay for tick-borne encephalitis virus (TBEV) point-of-care detection.","authors":"Alexander N Kudryavtsev, Eugenia E Denisova, Vasilisa V Krasitskaya, Ivan K Baykov, Nina V Tikunova, Ludmila A Frank","doi":"10.1007/s00216-025-06090-w","DOIUrl":"10.1007/s00216-025-06090-w","url":null,"abstract":"<p><p>Tick-borne encephalitis virus (TBEV), a highly pathogenic infectious agent that causes serious damage to the nervous system is mainly transmitted by Ixodidae ticks. The laboratory methods (immunoassay and the PCR-based one) are successfully used to detect the virus in tick samples thereby avoiding unwarranted immunoprophylaxis. However, there is a need to determine the tick infection outside the laboratory conditions. In this study, we have developed a one-stage (of mix-and-read type) method for detecting virus in biological samples based on split NanoLuc complementation assay. Artificial NanoLuc luciferase split fragments NLuc(N-residue), 17.6 kDa, and NLuc(C-residue), 11 a.a., were genetically fused with the protein prED3 (fragment of the TBEV capsid protein E) or mouse anti-TBEV single-chain antibody 14D5a in all possible variants. The corresponding hybrid proteins were synthesized in E. coli recombinant cells, purified and studied. Assembling of the luciferase fragments into a bioluminescent complex proceeded due to antigen-antibody affinity interaction. The most efficient luciferase complementation was observed for the pair 14D5a-NLucCter + prED3-NLucNter: the integral bioluminescence of the complex was 2.4% of that of the intact luciferase. Using this complex, a single-phase competitive enzyme immunoassay of TBEV-associated targets was developed. A large number of native ticks were analyzed and a statistically significant difference was shown between \"healthy\" and virus-carrying ticks. Lyophilized all-in-one reagents, reconstituted upon addition of the sample matrices, were developed and tested in model assay. The results offer a basis for the development of a point-of-need portable device for rapid tick detection.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5931-5939"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937829","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":"Asymmetric volume-mediated buffer control overcomes sensitivity limits in one-pot RAA-CRISPR/Cas12a visual detection.","authors":"Yue Zhang, Zunquan Zhao, Mingzhu Liu, Jincai Yang, Chun Yang, Nan Su, Jingran Sun, Yanjun Fang, Yonghui Wang, Xiaoli Li, Wang Chen, Jin Wu, Jialei Bai","doi":"10.1007/s00216-025-06095-5","DOIUrl":"10.1007/s00216-025-06095-5","url":null,"abstract":"<p><p>Rapid, low-cost, and visual nucleic acid detection methods are highly attractive for curbing colistin resistance spread through the food chain. CRISPR/Cas12a combined with recombinase-aided amplification (RAA) offers a one-pot, aerosol-free approach for visual detection. However, traditional one-pot systems often run Cas12a trans-cleavage in a buffer suitable for RAA, thus limiting Cas12a cleavage efficiency. This study proposes an asymmetric volume-optimized RAA-CRISPR/Cas12a assay for ultrasensitive visual detection of mobile colistin resistance gene mcr-1. Unlike conventional one-pot systems constrained by buffer incompatibility, our design spatially segregates a minimal-volume RAA-MIX (lid) from a CRISPR-dominant buffer microenvironment (tube bottom). This architecture leverages RAA's exponential amplification power to ensure sufficient product yield from minimal reaction volumes, while enabling subsequent enhancement of Cas12a trans-cleavage through automatic buffer assimilation upon mixing. The results were able to be visually observed under UV light, achieving 63.1% cost reduction compared to standard one-pot methods. The sensitivity of the proposed method for the mcr-1 gene was 2.5 copies/reaction, with anti-interference against other plasmids or bacteria. This method was applied to the detection of mcr-1 in animal-derived foods, showing satisfactory practical performance. By fundamentally reengineering buffer microenvironments through volume asymmetry, this work provides a general strategy for one-pot molecular diagnostics, achieving dual optimization of amplification and cleavage without trade-offs.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5971-5981"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145022564","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":"Speciation analysis of fungi by liquid atmospheric pressure MALDI mass spectrometry.","authors":"Lily R Adair, Ian M Jones, Rainer Cramer","doi":"10.1007/s00216-025-06094-6","DOIUrl":"10.1007/s00216-025-06094-6","url":null,"abstract":"<p><p>Fungal pathogens pose a growing threat to global health, necessitating rapid and accurate identification methods. Here, liquid atmospheric pressure matrix-assisted laser desorption/ionisation (LAP-MALDI) mass spectrometry (MS) is applied to fast lipid and protein profiling of Candida albicans and Saccharomyces cerevisiae from cultured colonies. Species-specific lipid profiles were observed in the m/z 600-1100 range, dominated by phospholipids as confirmed by tandem mass spectrometry (MS/MS). Following simple solid phase extraction clean-up, LAP-MALDI mass spectra revealed multiply charged protein ions suitable for MS/MS analysis. For C. albicans, the fully mature, species-specific WHS11 protein (~ 7 kDa; P43074) was detected intact and confidently identified by top-down MS/MS proteoform sequencing, including the cleavage of the N-terminal methionine initiator and the associated N-terminal acetylation. For S. cerevisiae, a set of proteoforms were sequenced by MS/MS analysis, which led to the identification of two species-specific proteins within the 'UniProtKB reference proteomes + Swiss-Prot' target database. One of these was also detected intact, and sequenced and identified as the fully mature HSP12 protein (~ 11.5 kDa; P22943). This work demonstrates the potential of LAP-MALDI MS and MS/MS biotyping as a powerful, label-free platform for rapid fungal classification and proteoform characterisation, offering substantial improvements over conventional MALDI biotyping.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5957-5969"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144999330","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":"Selection and characterization of novel DNA aptamers targeting colorectal cancer cells with malignant potential.","authors":"Yinuo Ma, Qun Wang, Shihan Sun, Yanxi Li, Rui Wang, Haihui Yang, Dian Wang, Yangyang Hao, Yujing Tong, Wanming Li","doi":"10.1007/s00216-025-06091-9","DOIUrl":"10.1007/s00216-025-06091-9","url":null,"abstract":"<p><p>Colorectal cancer (CRC) is associated with high morbidity and mortality rates worldwide. Therefore, early diagnosis and treatment are of great significance for improving the survival rate of CRC patients. Herein, we performed subtractive cell-SELEX using two cell lines (LS174T and CL187) with the same genetic background but different malignant potentials for the target and negative cells. Two aptamers LS1 and LS52 showed high affinity with target LS174T cells and high specificity for CRC cells with malignant potential, and exhibited good biological stability. In addition, we found that aptamer LS1 had the effect of promoting in vitro proliferation and monoclonal formation of CRC cells, and upregulated the expression of proliferation related proteins. Based on the high affinity of aptamer LS52, using LS52 as a molecular probe could effectively distinguish clinical CRC tissues and adjacent normal tissues, CRC metastatic tissues and non-metastatic tissues. Analysis of protein expression in magnetic bead sorted cells showed that the expression of malignant potential related proteins in LS52^high cells was higher than that in LS52^low cells. Finally, a cell membrane protein SRSF6 was identified as the target of aptamer LS52, which may serve as a new molecular target for CRC. Therefore, our work provides a systematic novel approach of studying potential biomarkers and a promising tool for cancer diagnosis and therapy.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5941-5955"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937983","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":"A ratiometric fluorescent aptamer assay based on RCA amplification mult-G-quadruplex.","authors":"Xinyu Xie, Weiling Li, Zhiguang Suo, Rui Guo, Min Wei, Huali Jin","doi":"10.1007/s00216-025-06083-9","DOIUrl":"10.1007/s00216-025-06083-9","url":null,"abstract":"<p><p>Ratiometric assay with dual signals can effectively avoid false positives of single signals. Therefore, a dual-signal assay was designed to detect Aflatoxin B1 (AFB1) using the affinity of the aptamers to the target as well as the binding force between the DNA strands. The first fluorescent signal was obtained by binding AFB1 to aptamer to shed the complementary sequence with the FAM (6-carboxyfluorescein). The G-quadruplex is a common DNA conformation capable of emitting light with special fluorescent dyes. At room temperature, the G-quadruplex is opened by the shed complementary strand to form double-stranded DNA, disrupting its original structure. The NMM (N-methylmesoporphyrin IX) dye is unable to embed the G-quadruplex and emit light. Taking advantage of the DNA conformational transition, NMM dye was introduced as the second fluorescence signal. Based on the above principle, a dual fluorescence signal ratiometric assay was constructed for the detection of AFB1 by the ratio of FAM and NMM fluorescence intensity. This study provides a prospective strategy for developing a dual-signal construct ratio assay.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5885-5893"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938213","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":"Carbon quantum dot-aptamer/MoS₂ nanosheet fluorescent sensor for ultrasensitive, noninvasive cortisol detection.","authors":"Rong Chen, Mingyu Wang, Hailiang Nie, Jinku Zhang, Hongyuan Yan","doi":"10.1007/s00216-025-06086-6","DOIUrl":"10.1007/s00216-025-06086-6","url":null,"abstract":"<p><p>This work presents the development of a highly sensitive, selective, and efficient aptamer-based fluorescent sensor for detecting cortisol in human urine. Carbon quantum dots-nucleic acid aptamer (CQDs-Apt) synthesized with excellent photoluminescent properties and stability, were selected as the fluorescent probe. In the presence of MoS₂-NSs, CQDs-Apt adsorbed onto the surface of MoS₂-NSs via electrostatic and π-π interactions, leading to strong and rapid fluorescence quenching due to static quenching mechanism between them. The CQDs-Apt/MoS₂-NSs complex can be employed as a \"turn-on\" fluorescent sensor for cortisol. Upon the addition of cortisol to the CQDs-Apt/MoS₂-NSs sensor, the aptamer specifically binds to cortisol, thereby weakening the interaction between CQDs-Apt and MoS₂-NSs. This results in the desorption of CDs-Apt from the surface of MoS₂-NSs and the recovery of the fluorescence signal. Under optimized conditions, the CQDs-Apt/MoS₂-NSs sensor exhibits a linear response to cortisol concentration (1-500 ng/mL) with a detection limit of 0.3 ng/mL. Furthermore, the sensor demonstrated excellent stability, high accuracy (92.0% to 97.7%), and superior precision (RSD ≤ 3.5%). This sensor has achieved a sensitive, rapid and efficient response to cortisol, and been successfully utilized for the detection of cortisol in human urine, demonstrating its potential for clinical cortisol detection.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5919-5930"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145022608","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":"Ferrocene-functionalized indium-oxo cluster nanozyme for the detection of hydrogen peroxide and ascorbic acid.","authors":"Shijie Wang, Yan Cheng, Zhelin Liu, Xiangting Dong, Bo Zhao","doi":"10.1007/s00216-025-06078-6","DOIUrl":"10.1007/s00216-025-06078-6","url":null,"abstract":"<p><p>Nanozymes are a class of nanomaterials with intrinsic enzyme-like properties, which overcome the limitations of natural enzymes, such as poor stability, high cost, and difficulty in storage, thus achieving rapid development. Herein, the enzyme-like activity of ferrocene-functionalized indium-oxo cluster (TPP⁺)[In₇FcDCA₆(µ₄-O^2-)₃(µ₃-OCH₃)(Cl^-)₃], named [In₇Fc₆], was analyzed. The obtained [In₇Fc₆] cluster has good peroxidase-like activity, which can catalyze the reaction of 3,3',5,5'-tetramethylbenzidine (TMB), o-phenylenediamine (OPD), and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) with H₂O₂. Therefore, based on the peroxidase-like activity of [In₇Fc₆], a colorimetric sensor for the detection of H₂O₂ was developed, which could detect H₂O₂ in the concentration range of 1-170 μM, and the detection limit was 0.24 μM. In addition, we also established a colorimetric sensor for detecting ascorbic acid (AA) in the range of 3-130 μM with a detection limit of 0.81 μM. The practicability of the established colorimetric sensor was also verified in cells and serum. This research opens up new horizons for exploring new application possibilities for clusters in biomolecular detection.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5847-5858"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937784","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":"Identification of serum N,N-dimethylglycine as a potential biomarker for prenatal diagnosis of congenital heart disease using ¹HNMR and UPLC-MS/MS metabonomics.","authors":"Baogang Xie, Dujuan Zhan, Le Wu, Lele Wang, Yongrong Lei, Meng Liu, Xiaodan Liu, Suping Li","doi":"10.1007/s00216-025-06084-8","DOIUrl":"10.1007/s00216-025-06084-8","url":null,"abstract":"<p><p>Congenital heart disease (CHD) poses significant clinical challenges due to limitations in early prenatal diagnosis. This study aimed to identify serum metabolic biomarkers for CHD using a combined metabolomics approach. Serum samples from 55 pregnant women carrying fetuses with confirmed CHD (CHDP group) and 49 healthy controls (ZCP group) were analyzed via non-targeted ¹HNMR metabolomics, revealing distinct metabolic profiles. Six choline pathway metabolites were further quantified by ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Among these, N,N-dimethylglycine (DMG) exhibited the most significant reduction in the CHDP group (1.07 ± 0.03 vs. 1.55 ± 0.04 µg/mL, p < 0.001), with an area under the ROC curve (AUROC) of 0.883 in the discovery cohort. Validation in an independent cohort (58 CHDP vs. 62 ZCP) confirmed DMG's diagnostic potential (AUROC = 0.818). While betaine-homocysteine methyltransferase 2 (BHMT2) activity showed no intergroup differences, DMG's consistent performance highlights its utility as a non-invasive biomarker. This study underscores the clinical value of metabonomics in prenatal CHD screening and establishes DMG as a promising diagnostic marker, potentially improving early detection and perinatal management.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5895-5906"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937961","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":"Comparative evaluation of phenyl isothiocyanate derivatization and \"dilute-and-shoot\" methods for HPLC-MS/MS-based targeted metabolomics analysis of amine-containing metabolites in plasma samples.","authors":"Kangkang Xu, Markus Aigensberger, Franz Berthiller, Heidi E Schwartz-Zimmermann","doi":"10.1007/s00216-025-06079-5","DOIUrl":"10.1007/s00216-025-06079-5","url":null,"abstract":"<p><p>Metabolomics, the study of small molecule metabolites in biological systems, is essential for disease diagnosis and biomarker discovery. A key consideration in developing targeted metabolomics methods using HPLC-MS/MS for human or animal plasma is whether to employ derivatization of amino acids, amino acid-related compounds, and biogenic amines. Derivatization with phenyl isothiocyanate (PITC) enhances ionization and LC-separation, but complicates sample preparation and introduces potential errors. This study focuses on (1) the validation of a PITC derivatization method employing reversed-phase (RP) HPLC-MS/MS analysis and (2) a comparison of two analytical approaches for targeted metabolomics analysis of animal and human plasma: the PITC derivatization-based RP-LC-MS/MS method and a \"dilute-and-shoot\" approach using hydrophilic interaction chromatography (HILIC)-MS/MS and RP-LC-MS/MS analysis. The derivatization method was validated for porcine plasma, assessing limits of detection, lower limits of quantification (LLOQs), linearity, repeatability, recovery, and trueness. Derivatization reduced LLOQs for derivatized compounds in pure solvent solutions but, due to higher dilution factors, resulted in similar LLOQs for derivatized compounds and higher LLOQs for non-derivatized compounds in plasma compared to the \"dilute-and-shoot\" method. Derivatization improved chromatographic separation of isomers and reduced carryover but introduced challenges such as matrix effects, coelution with impurities, and calibration issues. The \"dilute-and-shoot\" method performed better for non-derivatized compounds and was less error-prone. Both methods were applied to plasma from various species, demonstrating comparable concentrations for most metabolites. The results also emphasize the importance of using different approaches for cross-validation. Above all, this study highlights the strengths and limitations of both the derivatization method and the \"dilute-and-shoot\" approach, providing guidance for their application in targeted metabolomics.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5859-5872"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144938233","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 a novel method for generating Q-bodies using tyrosinase-mediated HA-tag labeling.","authors":"Hui-Seon Yun, Hanool Yun, Hee-Jin Jeong","doi":"10.1007/s00216-025-06087-5","DOIUrl":"10.1007/s00216-025-06087-5","url":null,"abstract":"<p><p>Quenchbodies (Q-bodies) are fluorescent antibodies that respond to antigen binding via a fluorescence quenching and de-quenching mechanism. To enhance the versatility of the generation method and expand the color range, we developed a novel Q-body generation approach using tyrosinase-mediated site-specific conjugation to a tyrosine-rich hemagglutinin (HA)-tag. A single-chain variable fragment (scFv) against programmed cell death-ligand 1 (PDL1) was engineered with an N-terminal HA-tag and expressed in Escherichia coli with high yield and purity. Site-specific conjugation of four hydrazide-functionalized dyes with different emission wavelengths was achieved using recombinant tyrosinase. The resulting Q-bodies were evaluated via fluorescence-linked immunosorbent assay, all of which demonstrated antigen concentration-dependent fluorescence enhancement. Notably, the tetramethylrhodamine (TAMRA)-labeled Q-body showed the highest signal-to-background ratio, limit of detection of 0.51 ± 0.01 μg/mL. This HA-tag-mediated Q-body generation strategy offers a robust and versatile tool for the production of Q-bodies with broad dye compatibility, enabling sensitive and multiplexed fluorescent immunoassays.</p>","PeriodicalId":462,"journal":{"name":"Analytical and Bioanalytical Chemistry","volume":" ","pages":"5785-5791"},"PeriodicalIF":3.8,"publicationDate":"2025-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144937831","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}