Pub Date : 2025-03-03DOI: 10.1016/j.biosx.2025.100592
Erika Alvarez Cañarte , Guilber Vergara Velez , Frank Guillermo Intriago Flor , Efrain Pérez Vega , Miguel Andrès Falconi Vèlez , Delia Noriega Verdugo , Génesis Pamela García García , Livis Sharith Díaz Alarcón , Andrés Miguel Anchundia Loor , Carlos Jadán-Piedra , Felipe Jadán Piedra
The energy and protein requirements of the population must be met, and the use of new analytical methods for rapid, low-cost detection of essential elements like lysine in reformulated foods is crucial. In this context, conditions were evaluated to develop a biosensor with lysine alpha oxidase (LOx), which showed high affinity for lysine with a KM of 0.32 mM. Different concentrations of cereals and legumes (70-30; 55-45; 85-15; quinoa-Lablab Purpureus; pole beans-Lablab Purpureus; and rye-Lablab Purpureus) were incorporated into meat sausages to enrich lysine, achieving a significant increase in lysine concentration (up to 75%) when 15% quinoa was substituted. The potentiometric signal, related to oxygen consumption during lysine oxidation, was detected at 15 s using a voltage of −600 mV. The biosensor, coupled with the immobilized enzyme, allowed the use of low volumes. A positive relationship was found between oxygen consumption (mg O2/L∗s-1) and lysine concentration in the range of 0.01–0.2 mM, with an R2 of 0.9964. The immobilized enzyme-based sensor demonstrated good sensitivity (0.01 mM) and the membrane could be reused up to 18 times, maintaining 92% of its initial activity after 70 days. The biosensor method showed minimal residue formation and had a strong correlation with high-performance liquid chromatography (HPLC) results, validating its accuracy.
{"title":"Cereals as sources of lysine in the reformulation of meat products. Evaluation using a biosensor","authors":"Erika Alvarez Cañarte , Guilber Vergara Velez , Frank Guillermo Intriago Flor , Efrain Pérez Vega , Miguel Andrès Falconi Vèlez , Delia Noriega Verdugo , Génesis Pamela García García , Livis Sharith Díaz Alarcón , Andrés Miguel Anchundia Loor , Carlos Jadán-Piedra , Felipe Jadán Piedra","doi":"10.1016/j.biosx.2025.100592","DOIUrl":"10.1016/j.biosx.2025.100592","url":null,"abstract":"<div><div>The energy and protein requirements of the population must be met, and the use of new analytical methods for rapid, low-cost detection of essential elements like lysine in reformulated foods is crucial. In this context, conditions were evaluated to develop a biosensor with lysine alpha oxidase (LOx), which showed high affinity for lysine with a K<sub>M</sub> of 0.32 mM. Different concentrations of cereals and legumes (70-30; 55-45; 85-15; quinoa-Lablab Purpureus; pole beans-Lablab Purpureus; and rye-Lablab Purpureus) were incorporated into meat sausages to enrich lysine, achieving a significant increase in lysine concentration (up to 75%) when 15% quinoa was substituted. The potentiometric signal, related to oxygen consumption during lysine oxidation, was detected at 15 s using a voltage of −600 mV. The biosensor, coupled with the immobilized enzyme, allowed the use of low volumes. A positive relationship was found between oxygen consumption (mg O<sub>2</sub>/L∗s-1) and lysine concentration in the range of 0.01–0.2 mM, with an R<sup>2</sup> of 0.9964. The immobilized enzyme-based sensor demonstrated good sensitivity (0.01 mM) and the membrane could be reused up to 18 times, maintaining 92% of its initial activity after 70 days. The biosensor method showed minimal residue formation and had a strong correlation with high-performance liquid chromatography (HPLC) results, validating its accuracy.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100592"},"PeriodicalIF":10.61,"publicationDate":"2025-03-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143548025","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.biosx.2025.100595
Alessio Sacco , Camilla Sacco Botto , Chiara D'Errico , Marina Ciuffo , Slavica Matić , Giulia Molinatto , Andrea M. Giovannozzi , Andrea M. Rossi , Emanuela Noris
Timely diagnosis of plant diseases and correct identification of etiological agents are fundamental to guarantee quality and quantity of agricultural products and food. Phytopathogenic bacteria induce devastating effects on crops. Their diagnosis and identification, mainly based on serological and molecular tools, are time consuming and expensive processes and require trained personnel. Among the innovative methods providing rapid, accurate, and reliable diagnosis at reduced costs, Raman spectroscopy (RS) is gathering considerable attention. RS provides a direct and non-destructive platform to gather information on the chemical and biochemical components of a sample, such as microorganism cultures, revealing their biological role. Due to the weak signals of bacterial cells in RS, a dielectrophoresis (DEP) approach was adopted to amplify the bacterial signals. Using Raman-DEP analysis, a dataset of spectra from different harmful phytopathogenic bacteria belonging to the genera Pseudomonas spp., Xanthomonas spp., and Erwinia spp. was obtained. Machine learning approaches were employed to discriminate isolates at the genus, species, and unprecedentedly at the pathovar level, reaching accuracies, precisions, recalls, and F1 scores of 94–100%. This approach offers important advancements in the non-destructive and rapid classification of microorganisms and is suitable to be readily extended to environmental and food diagnostics.
{"title":"Characterization of plant pathogenic bacteria at subspecies level using a dielectrophoresis device combined with Raman spectroscopy","authors":"Alessio Sacco , Camilla Sacco Botto , Chiara D'Errico , Marina Ciuffo , Slavica Matić , Giulia Molinatto , Andrea M. Giovannozzi , Andrea M. Rossi , Emanuela Noris","doi":"10.1016/j.biosx.2025.100595","DOIUrl":"10.1016/j.biosx.2025.100595","url":null,"abstract":"<div><div>Timely diagnosis of plant diseases and correct identification of etiological agents are fundamental to guarantee quality and quantity of agricultural products and food. Phytopathogenic bacteria induce devastating effects on crops. Their diagnosis and identification, mainly based on serological and molecular tools, are time consuming and expensive processes and require trained personnel. Among the innovative methods providing rapid, accurate, and reliable diagnosis at reduced costs, Raman spectroscopy (RS) is gathering considerable attention. RS provides a direct and non-destructive platform to gather information on the chemical and biochemical components of a sample, such as microorganism cultures, revealing their biological role. Due to the weak signals of bacterial cells in RS, a dielectrophoresis (DEP) approach was adopted to amplify the bacterial signals. Using Raman-DEP analysis, a dataset of spectra from different harmful phytopathogenic bacteria belonging to the genera <em>Pseudomonas</em> spp., <em>Xanthomonas</em> spp., and <em>Erwinia</em> spp. was obtained. Machine learning approaches were employed to discriminate isolates at the genus, species, and unprecedentedly at the pathovar level, reaching accuracies, precisions, recalls, and F1 scores of 94–100%. This approach offers important advancements in the non-destructive and rapid classification of microorganisms and is suitable to be readily extended to environmental and food diagnostics.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100595"},"PeriodicalIF":10.61,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143508556","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.biosx.2025.100594
Yahui Wen , Xinghai Wang , Jinxue Zhao , Xuejing Zhai , Wei Xia , Peiyi Li , Keqiang Lai , Lidong Wu
Enzymes are highly efficient catalysts in nature, governing countless reactions in biological systems. Enzyme-based hydrogels are three-dimensional network materials with enzyme activity, created by loading enzymes into hydrogels or using hydrogels to synthesize polymeric materials. Due to their high biocompatibility, enzyme-based hydrogels exhibit broad application potential in various fields. In recent years, researchers have been continuously exploring new preparation methods to improve the physical, chemical, and biological properties of enzyme-based hydrogels. Additionally, the introduction of materials such as graphene, carbon nanotubes, and metal-organic frameworks (MOFs) has further enhanced the biological activity of these hydrogels. This review summarizes recent preparation methods and structural characteristics of enzyme-based hydrogels, with a focus on their applications in biosensors and medical fields, highlighting the vast application prospects of enzyme-based hydrogels.
{"title":"Preparation and application of enzyme-based hydrogels","authors":"Yahui Wen , Xinghai Wang , Jinxue Zhao , Xuejing Zhai , Wei Xia , Peiyi Li , Keqiang Lai , Lidong Wu","doi":"10.1016/j.biosx.2025.100594","DOIUrl":"10.1016/j.biosx.2025.100594","url":null,"abstract":"<div><div>Enzymes are highly efficient catalysts in nature, governing countless reactions in biological systems. Enzyme-based hydrogels are three-dimensional network materials with enzyme activity, created by loading enzymes into hydrogels or using hydrogels to synthesize polymeric materials. Due to their high biocompatibility, enzyme-based hydrogels exhibit broad application potential in various fields. In recent years, researchers have been continuously exploring new preparation methods to improve the physical, chemical, and biological properties of enzyme-based hydrogels. Additionally, the introduction of materials such as graphene, carbon nanotubes, and metal-organic frameworks (MOFs) has further enhanced the biological activity of these hydrogels. This review summarizes recent preparation methods and structural characteristics of enzyme-based hydrogels, with a focus on their applications in biosensors and medical fields, highlighting the vast application prospects of enzyme-based hydrogels.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100594"},"PeriodicalIF":10.61,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143508555","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-20DOI: 10.1016/j.biosx.2025.100596
Omar Ramadan, Rabeay Y.A. Hassan
Breast cancer patients often exhibit positive expression of selective tumor biomarkers including the carcinoembryonic antigen (CEA), and human epidermal growth factor receptor 2 (HER2). These potential protein biomarkers are highly crucial for the selective and early tumor diagnosis. In this study, disposable dual impedimetric immunosensing systems were constructed using the cobalt molybdate (CoMoO4) and polyaniline/polypyrrole (PANI-PPy) nanocomposite, for the rapid quantification of CEA and HER2 concentration levels. The designed sensing probes were fabricated by chemically conjugating two-targeting antibodies (anti-CEA and anti-HER2) on the nanostructured electrode surfaces modified with (AuNPs/CoMoO4@PANI-PPy). These successful chemical antibody immobilizations were enabled by the use of 4-aminothiophenol (4-ATP) as a cross-linking agent. The impedimetric immunosensing assay was fully optimized by screening of nanomaterials, determining the optimal ratio of the nanocomposite, adjusting the time of self-assembled monolayer (SAM) formation, and identifying the degree of cross-reactivity with potential non-targeting molecules. Accordingly, the biosensors demonstrated distinguished linear dynamic ranges of 1.0 fg/mL to 100 ng/mL for the CEA and 25 fg/mL to 100 ng/mL for the HER2, with detection limits of 8.2 fg/mL and 4.9 fg/mL for CEA and HER2, respectively. The immunesensor's reproducibility and stability were studied, showing minimal variation in charge transfer resistance (RSD = 1.33%, n = 5) and maintaining 82.5% functionality after four weeks of storage at 4 °C. These findings highlighted the potential use of these biosensor chips for clinical applications in early breast cancer detection and rapid diagnosis.
{"title":"Nanostructured immunosensing system for label-free impedimetric detection of multiple breast cancer biomarkers (CEA and HER2) using CoMoO4@PANI-PPy Nanocomposite","authors":"Omar Ramadan, Rabeay Y.A. Hassan","doi":"10.1016/j.biosx.2025.100596","DOIUrl":"10.1016/j.biosx.2025.100596","url":null,"abstract":"<div><div>Breast cancer patients often exhibit positive expression of selective tumor biomarkers including the carcinoembryonic antigen (CEA), and human epidermal growth factor receptor 2 (HER2). These potential protein biomarkers are highly crucial for the selective and early tumor diagnosis. In this study, disposable dual impedimetric immunosensing systems were constructed using the cobalt molybdate (CoMoO<sub>4</sub>) and polyaniline/polypyrrole (PANI-PPy) nanocomposite, for the rapid quantification of CEA and HER2 concentration levels. The designed sensing probes were fabricated by chemically conjugating two-targeting antibodies (anti-CEA and anti-HER2) on the nanostructured electrode surfaces modified with (AuNPs/CoMoO<sub>4</sub>@PANI-PPy). These successful chemical antibody immobilizations were enabled by the use of 4-aminothiophenol (4-ATP) as a cross-linking agent. The impedimetric immunosensing assay was fully optimized by screening of nanomaterials, determining the optimal ratio of the nanocomposite, adjusting the time of self-assembled monolayer (SAM) formation, and identifying the degree of cross-reactivity with potential non-targeting molecules. Accordingly, the biosensors demonstrated distinguished linear dynamic ranges of 1.0 fg/mL to 100 ng/mL for the CEA and 25 fg/mL to 100 ng/mL for the HER2, with detection limits of 8.2 fg/mL and 4.9 fg/mL for CEA and HER2, respectively. The immunesensor's reproducibility and stability were studied, showing minimal variation in charge transfer resistance (RSD = 1.33%, n = 5) and maintaining 82.5% functionality after four weeks of storage at 4 °C. These findings highlighted the potential use of these biosensor chips for clinical applications in early breast cancer detection and rapid diagnosis.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100596"},"PeriodicalIF":10.61,"publicationDate":"2025-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143479828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-17DOI: 10.1016/j.biosx.2025.100593
Dianna J. Rowe , Timothy A. Khalil , Michael N. Kammer , Caroline M. Godfrey , Yong Zou , Cindy L. Vnencak-Jones , David Xiao , Stephen Deppen , Eric L. Grogan
Studies show CYFRA 21-1 fragments of cytokeratin 19 (CK19) to be promising biomarkers for non-small cell lung cancer (NSCLC). Although previous literature identifies specific CYFRA 21-1 antibody binding epitopes, the exact molecular weight of the CK19 fragment being detected by current assays is not well-documented. Serum samples from 58 patients (lung cancer (N = 36), control (N = 22)) were used to measure CYFRA 21-1 across four different quantification assays: enzyme-linked immunosorbent assay (ELISA), chemiluminescent assay (ChLIA), electrochemiluminescence immunoassay (ECLIA), and compensated interferometric reader (CIR). In the cancer group, correlation between ECLIA and ELISA was high (R(Pearson) = 0.948, r(Spearman) = 0.868) while correlation between ECLIA vs ChLIA and ECLIA vs CIR was low (R= 0.005, r = −0.0593), (R = 0.0275, r = 0.167), respectively. In the control group, correlation between ECLIA and ELISA was high (R = 0.948, r = 0.868) while correlation between ECLIA vs ChLIA and ECLIA vs CIR was low (R = 0.005, r = −0.0593), (R = 0.0275, r = 0.167), respectively. Compared to ECLIA, concordance coefficients (pc) were poor (pc < 0.90) across all assays except for cancers group in ELISA (pc = 0.913). ECLIA was the only assay to report control ranges above 1 ng/mL CYFRA 21-1 (ECLIA, 1.14–21.59 ng/mL; ELISA, 0.79–24.26 ng/mL; ChLIA, 0.062–0.691 ng/mL; 0.08–7.68 ng/mL). Differing sizes of the protein being measured by each assay may have a role in the discrepancies observed. Given the different CYFRA 21-1 concentration estimates among assays, further characterization of the fragment and its release during epithelial malignancies, such as NSCLC, is imperative to developing effective biomarker assays.
{"title":"A deeper evaluation of cytokeratin fragment 21-1 as a lung cancer tumor marker and comparison of different assays","authors":"Dianna J. Rowe , Timothy A. Khalil , Michael N. Kammer , Caroline M. Godfrey , Yong Zou , Cindy L. Vnencak-Jones , David Xiao , Stephen Deppen , Eric L. Grogan","doi":"10.1016/j.biosx.2025.100593","DOIUrl":"10.1016/j.biosx.2025.100593","url":null,"abstract":"<div><div>Studies show CYFRA 21-1 fragments of cytokeratin 19 (CK19) to be promising biomarkers for non-small cell lung cancer (NSCLC). Although previous literature identifies specific CYFRA 21-1 antibody binding epitopes, the exact molecular weight of the CK19 fragment being detected by current assays is not well-documented. Serum samples from 58 patients (lung cancer (N = 36), control (N = 22)) were used to measure CYFRA 21-1 across four different quantification assays: enzyme-linked immunosorbent assay (ELISA), chemiluminescent assay (ChLIA), electrochemiluminescence immunoassay (ECLIA), and compensated interferometric reader (CIR). In the cancer group, correlation between ECLIA and ELISA was high (R<em>(Pearson)</em> = 0.948, r<em>(Spearman)</em> = 0.868) while correlation between ECLIA vs ChLIA and ECLIA vs CIR was low (R<em>=</em> 0.005, r = −0.0593), (R = 0.0275, r = 0.167), respectively. In the control group, correlation between ECLIA and ELISA was high (R = 0.948, r = 0.868) while correlation between ECLIA vs ChLIA and ECLIA vs CIR was low (R = 0.005, r = −0.0593), (R = 0.0275, r = 0.167), respectively. Compared to ECLIA, concordance coefficients <strong>(</strong><em>p</em><sub><em>c</em></sub>) were poor (<em>p</em><sub><em>c</em></sub> < 0.90) across all assays except for cancers group in ELISA (<em>p</em><sub><em>c</em></sub> = 0.913). ECLIA was the only assay to report control ranges above 1 ng/mL CYFRA 21-1 (ECLIA, 1.14–21.59 ng/mL; ELISA, 0.79–24.26 ng/mL; ChLIA, 0.062–0.691 ng/mL; 0.08–7.68 ng/mL). Differing sizes of the protein being measured by each assay may have a role in the discrepancies observed. Given the different CYFRA 21-1 concentration estimates among assays, further characterization of the fragment and its release during epithelial malignancies, such as NSCLC, is imperative to developing effective biomarker assays.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100593"},"PeriodicalIF":10.61,"publicationDate":"2025-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143479829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-16DOI: 10.1016/j.biosx.2025.100585
Taekmin Kim, Dayeon Choi, Soonjoung Kwon, Se-Eun Park, Jongki Kim, Yoon-Ho Lee, Seung-Cheol Choi, Se-Hwan Paek
We developed a novel cancer detection method that leverages changes in local exosome heterogeneity to regulate bulk-scale heterogeneity in body fluids. Using an automated, reversible isolation system and downstream immunoassays, we investigated exosome subclasses exhibiting marker composition changes associated with cancer. We introduced an index, R, to measure local heterogeneity changes, quantifying the proportion of a third marker within double-marker-positive subclasses. This method, which employs pan-exosome tetraspanins CD9, CD63, and CD81, revealed distinct heterogeneity differences between normal and cancer samples. These differences were consistently demonstrated in exosome samples derived from various cell lines and human sera. Notably, in clinical samples, R values distributed samples from healthy donors and cancer patients into unique patterns based on the third marker within specific subclasses. However, across subclasses, R value changes were significantly smaller in healthy samples than in cancer samples. Sorting patterns were simulated using exosomes derived from immune cells, indicating an immune response contribution to the observed signals. These findings highlight the method's potential for identifying biomarkers for specific cancer diagnoses and multi-cancer screening.
我们开发了一种新型癌症检测方法，它利用局部外泌体异质性的变化来调节体液中大量的异质性。利用自动化、可逆分离系统和下游免疫测定，我们研究了表现出与癌症相关的标记物组成变化的外泌体亚类。我们引入了一个指数 R 来衡量局部异质性变化，量化双标记物阳性亚类中第三个标记物的比例。这种方法采用了泛外显子四聚体 CD9、CD63 和 CD81，揭示了正常样本和癌症样本之间明显的异质性差异。这些差异在来自不同细胞系和人类血清的外泌体样本中得到了一致证实。值得注意的是，在临床样本中，根据特定亚类中的第三个标记物，R 值将来自健康供体和癌症患者的样本分布成独特的模式。不过，在不同的亚类中，健康样本的 R 值变化明显小于癌症样本。使用免疫细胞的外泌体模拟了分选模式，表明免疫反应对观察到的信号有贡献。这些发现凸显了该方法在确定用于特定癌症诊断和多种癌症筛查的生物标记物方面的潜力。
{"title":"A liquid biopsy algorithm exploiting local exosome heterogeneity","authors":"Taekmin Kim, Dayeon Choi, Soonjoung Kwon, Se-Eun Park, Jongki Kim, Yoon-Ho Lee, Seung-Cheol Choi, Se-Hwan Paek","doi":"10.1016/j.biosx.2025.100585","DOIUrl":"10.1016/j.biosx.2025.100585","url":null,"abstract":"<div><div>We developed a novel cancer detection method that leverages changes in local exosome heterogeneity to regulate bulk-scale heterogeneity in body fluids. Using an automated, reversible isolation system and downstream immunoassays, we investigated exosome subclasses exhibiting marker composition changes associated with cancer. We introduced an index, R, to measure local heterogeneity changes, quantifying the proportion of a third marker within double-marker-positive subclasses. This method, which employs pan-exosome tetraspanins CD9, CD63, and CD81, revealed distinct heterogeneity differences between normal and cancer samples. These differences were consistently demonstrated in exosome samples derived from various cell lines and human sera. Notably, in clinical samples, R values distributed samples from healthy donors and cancer patients into unique patterns based on the third marker within specific subclasses. However, across subclasses, R value changes were significantly smaller in healthy samples than in cancer samples. Sorting patterns were simulated using exosomes derived from immune cells, indicating an immune response contribution to the observed signals. These findings highlight the method's potential for identifying biomarkers for specific cancer diagnoses and multi-cancer screening.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100585"},"PeriodicalIF":10.61,"publicationDate":"2025-02-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143445300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-10DOI: 10.1016/j.biosx.2025.100589
Mahtab Kokabi , Gulam M. Rather , Mehdi Javanmard
In this study, we employed a custom microfluidic sensor to measure the electrical impedance of cancer cells, aiming to differentiate between suspension and adherent phenotypes. We investigated three suspension cell lines—JeKo-1, MM-1R, and Maver-1—representing hematological malignancies, and two adherent breast cancer cell lines—MDA-MB-231 and MDA- MB-468. The impedance measurements revealed significant differences corresponding to the cells’ growth patterns. Suspension cells exhibited lower median impedance values compared to adherent cells, likely due to variations in cell size, complexity, and membrane properties. Suspension cells, being smaller and less structurally complex, demonstrated reduced impedance, whereas adherent cells, which are larger and form stronger surface attachments, displayed higher impedance values. These findings highlight the potential of electrical impedance as a tool for distinguishing cancer cell types. To further validate these results, we propose focusing on the transition of a single cancer cell type between adherent and suspension states, simulating in vitro cancer proliferation conditions. This approach will provide a deeper understanding of how electrical properties evolve during this transition and may offer insights into distinguishing between benign and metastatic cancers or assessing metastatic stages. The observed reduction in impedance during the adherent-to-suspension transition supports the potential diagnostic utility of this method. Overall, this study demonstrates that electrical impedance provides a rapid, label-free approach for enhancing cancer cell diagnostics based on distinct electrical characteristics.
{"title":"Microfluidic impedance sensing distinguishes cancer cell states: A step towards point-of-care diagnostics","authors":"Mahtab Kokabi , Gulam M. Rather , Mehdi Javanmard","doi":"10.1016/j.biosx.2025.100589","DOIUrl":"10.1016/j.biosx.2025.100589","url":null,"abstract":"<div><div>In this study, we employed a custom microfluidic sensor to measure the electrical impedance of cancer cells, aiming to differentiate between suspension and adherent phenotypes. We investigated three suspension cell lines—JeKo-1, MM-1R, and Maver-1—representing hematological malignancies, and two adherent breast cancer cell lines—MDA-MB-231 and MDA- MB-468. The impedance measurements revealed significant differences corresponding to the cells’ growth patterns. Suspension cells exhibited lower median impedance values compared to adherent cells, likely due to variations in cell size, complexity, and membrane properties. Suspension cells, being smaller and less structurally complex, demonstrated reduced impedance, whereas adherent cells, which are larger and form stronger surface attachments, displayed higher impedance values. These findings highlight the potential of electrical impedance as a tool for distinguishing cancer cell types. To further validate these results, we propose focusing on the transition of a single cancer cell type between adherent and suspension states, simulating in vitro cancer proliferation conditions. This approach will provide a deeper understanding of how electrical properties evolve during this transition and may offer insights into distinguishing between benign and metastatic cancers or assessing metastatic stages. The observed reduction in impedance during the adherent-to-suspension transition supports the potential diagnostic utility of this method. Overall, this study demonstrates that electrical impedance provides a rapid, label-free approach for enhancing cancer cell diagnostics based on distinct electrical characteristics.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100589"},"PeriodicalIF":10.61,"publicationDate":"2025-02-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143403226","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-07DOI: 10.1016/j.biosx.2025.100591
Gabriel López-Peña , Ana Pallarés Vilar , Aida Jaafar , Silvia Simón-Fuente , Antonio Arranz , Maria Ribagorda , Miguel Manso-Silván , Emma Martín Rodríguez
This study presents a novel noninvasive and enzyme-free dual sensor based on porous silicon (PSi) for the detection of D-glucose through tear fluid analysis. The sensor incorporates NaGdF4:20% Yb3+, 2% Er3+ nanoparticles (YbEr-NPs) immobilized on a PSi substrate functionalized with a phenylboronic ester molecule. The YbEr-NPs enable luminescence-based sensing, exploiting the sensitivity of Er3+ ions to the OH vibrational groups present in D-glucose. Electrodes fabricated on a PSi substrate allow for voltammetric analysis, providing a second detection method. The voltammetric sensor achieved a limit of detection (LoD) of 20 mg/dL in the range of 20–200 mg/dL, which is suitable for detecting D-glucose concentrations in the tear fluid of diabetic patients. The luminescence-based sensor, utilizing the red-to-green emission ratio of YbEr-NPs, reached a LoD of 140 mg/dL in the range of 10–70 mg/dL, covering the hyperglycemic range. The interaction between the YbEr-NPs and the PSi substrate led to the appearance of new emission bands and increased intensity, attributed to surface defects acting as an additional excitation source. The 556 nm emission band showed a strong dependence on the D-glucose concentration, improving the LoD to 110 mg/dL. This provides a novel strategy for D-glucose detection based on the effect of the molecule on the interaction between the Rare-earth-doped nanoparticles and the PSi substrate. This triple-sensing approach offers a promising solution for noninvasive glucose monitoring, enabling detection in both the hypoglycemic and hyperglycemic ranges.
{"title":"Integrating voltammetry and substrate-enhanced luminescence for noninvasive glucose sensing","authors":"Gabriel López-Peña , Ana Pallarés Vilar , Aida Jaafar , Silvia Simón-Fuente , Antonio Arranz , Maria Ribagorda , Miguel Manso-Silván , Emma Martín Rodríguez","doi":"10.1016/j.biosx.2025.100591","DOIUrl":"10.1016/j.biosx.2025.100591","url":null,"abstract":"<div><div>This study presents a novel noninvasive and enzyme-free dual sensor based on porous silicon (PSi) for the detection of D-glucose through tear fluid analysis. The sensor incorporates NaGdF<sub>4</sub>:20% Yb<sup>3+</sup>, 2% Er<sup>3+</sup> nanoparticles (YbEr-NPs) immobilized on a PSi substrate functionalized with a phenylboronic ester molecule. The YbEr-NPs enable luminescence-based sensing, exploiting the sensitivity of Er<sup>3+</sup> ions to the OH vibrational groups present in D-glucose. Electrodes fabricated on a PSi substrate allow for voltammetric analysis, providing a second detection method. The voltammetric sensor achieved a limit of detection (LoD) of 20 mg/dL in the range of 20–200 mg/dL, which is suitable for detecting D-glucose concentrations in the tear fluid of diabetic patients. The luminescence-based sensor, utilizing the red-to-green emission ratio of YbEr-NPs, reached a LoD of 140 mg/dL in the range of 10–70 mg/dL, covering the hyperglycemic range. The interaction between the YbEr-NPs and the PSi substrate led to the appearance of new emission bands and increased intensity, attributed to surface defects acting as an additional excitation source. The 556 nm emission band showed a strong dependence on the D-glucose concentration, improving the LoD to 110 mg/dL. This provides a novel strategy for D-glucose detection based on the effect of the molecule on the interaction between the Rare-earth-doped nanoparticles and the PSi substrate. This triple-sensing approach offers a promising solution for noninvasive glucose monitoring, enabling detection in both the hypoglycemic and hyperglycemic ranges.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100591"},"PeriodicalIF":10.61,"publicationDate":"2025-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143421541","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-02-06DOI: 10.1016/j.biosx.2025.100590
Pierre Guermonprez , Le Tu Anh , Louis Renaud , Pierre Nioche , Eric Krejci , Aurélie Alleaume-Butaux , Nicolas Battaglini , Vu Thi Thu , Sébastien Sanaur , Benoît Piro
Detection of Ribonucleic acids (RNA) is a critical step in the identification of viral or bacterial infections in humans and animals. Reverse transcriptase-polymerase chain reaction (RT-PCR) remains the gold standard, but clustered regularly interspaced short palindromic repeats linked to a Cas endoribonuclease particle (CRISPR/Cas) have recently revolutionized the recognition step of two RNAs, i.e. the CRISPR-RNA (crRNA) and the target, providing a much better selectivity compared to the naked hybridization on which RT-PCR is based. Here, we combine the high efficiency of the CRISPR/Cas13a system with the transduction and amplification capabilities of an electrolyte-gated graphene field-effect transistor (EGGFET) for the detection of specific RNA sequences. In these devices, fabricated on flexible plastic substrates, the active material (reduced graphene oxide, rGO) is deposited by printing and then functionalized with Au nanoparticles decorated with polyU RNA reporter strands. In this system, the CRISPR/Cas13a complex acts as a catalyst: in the presence of a specific RNA target sequence, the enzymatic function is activated and the polyU RNA reporter strands are cleaved from the Au nanoparticles, inducing a loss of negative charges on the rGO layer. This phenomenon leads to measurable changes in the transfer curve of the transistors. These sensors were tested for the detection of a SARS-CoV-2 RNA sequence and showed a linear response in the range of 10−7 - 102 ng μL−1 with an estimated limit of detection of 10 fM. This work is an important milestone in the development of the next generation of point-of-care RNA sensors.
{"title":"An electrolyte-gated transistor for the monitoring of a CRISPR/Cas activity","authors":"Pierre Guermonprez , Le Tu Anh , Louis Renaud , Pierre Nioche , Eric Krejci , Aurélie Alleaume-Butaux , Nicolas Battaglini , Vu Thi Thu , Sébastien Sanaur , Benoît Piro","doi":"10.1016/j.biosx.2025.100590","DOIUrl":"10.1016/j.biosx.2025.100590","url":null,"abstract":"<div><div>Detection of Ribonucleic acids (RNA) is a critical step in the identification of viral or bacterial infections in humans and animals. Reverse transcriptase-polymerase chain reaction (RT-PCR) remains the gold standard, but clustered regularly interspaced short palindromic repeats linked to a Cas endoribonuclease particle (CRISPR/Cas) have recently revolutionized the recognition step of two RNAs, i.e. the CRISPR-RNA (crRNA) and the target, providing a much better selectivity compared to the naked hybridization on which RT-PCR is based. Here, we combine the high efficiency of the CRISPR/Cas13a system with the transduction and amplification capabilities of an electrolyte-gated graphene field-effect transistor (EGGFET) for the detection of specific RNA sequences. In these devices, fabricated on flexible plastic substrates, the active material (reduced graphene oxide, rGO) is deposited by printing and then functionalized with Au nanoparticles decorated with polyU RNA reporter strands. In this system, the CRISPR/Cas13a complex acts as a catalyst: in the presence of a specific RNA target sequence, the enzymatic function is activated and the polyU RNA reporter strands are cleaved from the Au nanoparticles, inducing a loss of negative charges on the rGO layer. This phenomenon leads to measurable changes in the transfer curve of the transistors. These sensors were tested for the detection of a SARS-CoV-2 RNA sequence and showed a linear response in the range of 10<sup>−7</sup> - 10<sup>2</sup> ng μL<sup>−1</sup> with an estimated limit of detection of 10 fM. This work is an important milestone in the development of the next generation of point-of-care RNA sensors.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100590"},"PeriodicalIF":10.61,"publicationDate":"2025-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143377878","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-01-31DOI: 10.1016/j.biosx.2025.100588
Nedim Hacıosmanoğlu , Murat Alp Güngen , Eylul Gulsen Yilmaz , Emre Ece , Alphan Uzun , Arda Taşcan , Burak M. Görmüş , Ismail Eş , Fatih Inci
Three-dimensional (3D) cell cultures, especially spheroids, provide a physiologically accurate model for cancer research in comparison to conventional two-dimensional (2D) cultures. Nevertheless, the difficulties in producing and analyzing spheroids have impeded their extensive use in high-throughput screening—a critical process for drug discovery. This study presents a simplified process for the effective creation and examination of spheroids using a 3D-printed mold casted polydimethylsiloxane (PDMS) microwells. The utilization of our specially constructed mold facilitated the creation of consistent spheroids, which were subsequently exposed to doxorubicin for the purpose of anticancer medication treatment. We herein improved spheroid analysis by including a convolutional neural network (CNN) model, specifically U-Net, into a graphical user interface (GUI). This integration allows for automated detection and measurement of spheroid size from microscope pictures. The performance of this system surpassed that of conventional image analysis methods in terms of both accuracy and efficiency. The implemented workflow offers a scalable and cost-efficient platform for conducting high-throughput drug screening, which has the potential to enhance the success rates of cancer therapies in clinical trials.
{"title":"SpAi: A machine-learning supported experimental workflow for high-throughput spheroid production and analysis","authors":"Nedim Hacıosmanoğlu , Murat Alp Güngen , Eylul Gulsen Yilmaz , Emre Ece , Alphan Uzun , Arda Taşcan , Burak M. Görmüş , Ismail Eş , Fatih Inci","doi":"10.1016/j.biosx.2025.100588","DOIUrl":"10.1016/j.biosx.2025.100588","url":null,"abstract":"<div><div>Three-dimensional (3D) cell cultures, especially spheroids, provide a physiologically accurate model for cancer research in comparison to conventional two-dimensional (2D) cultures. Nevertheless, the difficulties in producing and analyzing spheroids have impeded their extensive use in high-throughput screening—a critical process for drug discovery. This study presents a simplified process for the effective creation and examination of spheroids using a 3D-printed mold casted polydimethylsiloxane (PDMS) microwells. The utilization of our specially constructed mold facilitated the creation of consistent spheroids, which were subsequently exposed to doxorubicin for the purpose of anticancer medication treatment. We herein improved spheroid analysis by including a convolutional neural network (CNN) model, specifically U-Net, into a graphical user interface (GUI). This integration allows for automated detection and measurement of spheroid size from microscope pictures. The performance of this system surpassed that of conventional image analysis methods in terms of both accuracy and efficiency. The implemented workflow offers a scalable and cost-efficient platform for conducting high-throughput drug screening, which has the potential to enhance the success rates of cancer therapies in clinical trials.</div></div>","PeriodicalId":260,"journal":{"name":"Biosensors and Bioelectronics: X","volume":"23 ","pages":"Article 100588"},"PeriodicalIF":10.61,"publicationDate":"2025-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143421542","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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