Ruma Paul, Yermary Morales-Lozada, Brian J. Sánchez Colón, Andrea R. Hernandez, Sourav Roy and Carlos R. Cabrera*,
{"title":"基于血液的生物标志物CCSP-2的结直肠癌无标记障碍免疫传感器","authors":"Ruma Paul, Yermary Morales-Lozada, Brian J. Sánchez Colón, Andrea R. Hernandez, Sourav Roy and Carlos R. Cabrera*, ","doi":"10.1021/acsmeasuresciau.4c0007310.1021/acsmeasuresciau.4c00073","DOIUrl":null,"url":null,"abstract":"<p >Colorectal cancer (CRC) is one of the most treatable cancers, yet it ranks second in mortality worldwide. Early detection significantly impacts treatment outcomes, but early stage CRC often presents no symptoms or nonspecific symptoms. The current screening methods are invasive and lacks specificity, hindering widespread CRC screening efforts. This underscores the urgent need for improved CRC screening tools. In this study, a label-free impedimetric immunosensor for detecting colon cancer-secreted protein-2 (CCSP-2), which exhibits a mean 78-fold increase in primary colon cancers compared to normal mucosa, was developed. Our cost-effective and noninvasive electrochemical immunosensor for CCSP-2 biomarker detection aims to facilitate early diagnosis and monitoring of CRC. The designed immunosensor features a functionalized gold electrode (Au) modified with cysteine-modified recombinant protein G (RPGCys) to immobilize the CCSP-2 antibody (Ab), and bovine serum albumin (BSA) used to prevent sensor surface fouling. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were employed to analyze the electrochemical response to the binding of CCSP-2 antigen (Ag) and Ab. The changes in relative charge transfer resistance (Δ<i>R</i><sub>ct</sub>/<i>R</i><sub>cti</sub>) with varying concentrations of Ag were plotted and a calibration curve was established between Δ<i>R</i><sub>ct</sub>/<i>R</i><sub>cti</sub> and logarithm of Ag concentration to assess sensor’s sensitivity. The sensor demonstrated a linear response (<i>R</i><sup>2</sup> = 0.95) within the range of 10–100 ng/μL, plateauing after 100 ng/μL, with a detection limit of 0.71 ng/μL. Statistical analysis of specificity and selectivity studies showed significant differences in Ag detection compared to blank and nonspecific protein BSA, both with and without cell extracts. This immunosensor effectively detects the CRC biomarker CCSP-2 with high sensitivity and specificity. Integrating this sensor with other sensors for serum CRC biomarkers present a promising approach for developing diagnostic and prognostic tools for CRC.</p>","PeriodicalId":29800,"journal":{"name":"ACS Measurement Science Au","volume":"5 1","pages":"87–95 87–95"},"PeriodicalIF":4.6000,"publicationDate":"2025-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://pubs.acs.org/doi/epdf/10.1021/acsmeasuresciau.4c00073","citationCount":"0","resultStr":"{\"title\":\"Colorectal Cancer Label-Free Impedimetric Immunosensor for Blood-Based Biomarker CCSP-2\",\"authors\":\"Ruma Paul, Yermary Morales-Lozada, Brian J. Sánchez Colón, Andrea R. Hernandez, Sourav Roy and Carlos R. Cabrera*, \",\"doi\":\"10.1021/acsmeasuresciau.4c0007310.1021/acsmeasuresciau.4c00073\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p >Colorectal cancer (CRC) is one of the most treatable cancers, yet it ranks second in mortality worldwide. Early detection significantly impacts treatment outcomes, but early stage CRC often presents no symptoms or nonspecific symptoms. The current screening methods are invasive and lacks specificity, hindering widespread CRC screening efforts. This underscores the urgent need for improved CRC screening tools. In this study, a label-free impedimetric immunosensor for detecting colon cancer-secreted protein-2 (CCSP-2), which exhibits a mean 78-fold increase in primary colon cancers compared to normal mucosa, was developed. Our cost-effective and noninvasive electrochemical immunosensor for CCSP-2 biomarker detection aims to facilitate early diagnosis and monitoring of CRC. The designed immunosensor features a functionalized gold electrode (Au) modified with cysteine-modified recombinant protein G (RPGCys) to immobilize the CCSP-2 antibody (Ab), and bovine serum albumin (BSA) used to prevent sensor surface fouling. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were employed to analyze the electrochemical response to the binding of CCSP-2 antigen (Ag) and Ab. The changes in relative charge transfer resistance (Δ<i>R</i><sub>ct</sub>/<i>R</i><sub>cti</sub>) with varying concentrations of Ag were plotted and a calibration curve was established between Δ<i>R</i><sub>ct</sub>/<i>R</i><sub>cti</sub> and logarithm of Ag concentration to assess sensor’s sensitivity. The sensor demonstrated a linear response (<i>R</i><sup>2</sup> = 0.95) within the range of 10–100 ng/μL, plateauing after 100 ng/μL, with a detection limit of 0.71 ng/μL. Statistical analysis of specificity and selectivity studies showed significant differences in Ag detection compared to blank and nonspecific protein BSA, both with and without cell extracts. This immunosensor effectively detects the CRC biomarker CCSP-2 with high sensitivity and specificity. 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Colorectal Cancer Label-Free Impedimetric Immunosensor for Blood-Based Biomarker CCSP-2
Colorectal cancer (CRC) is one of the most treatable cancers, yet it ranks second in mortality worldwide. Early detection significantly impacts treatment outcomes, but early stage CRC often presents no symptoms or nonspecific symptoms. The current screening methods are invasive and lacks specificity, hindering widespread CRC screening efforts. This underscores the urgent need for improved CRC screening tools. In this study, a label-free impedimetric immunosensor for detecting colon cancer-secreted protein-2 (CCSP-2), which exhibits a mean 78-fold increase in primary colon cancers compared to normal mucosa, was developed. Our cost-effective and noninvasive electrochemical immunosensor for CCSP-2 biomarker detection aims to facilitate early diagnosis and monitoring of CRC. The designed immunosensor features a functionalized gold electrode (Au) modified with cysteine-modified recombinant protein G (RPGCys) to immobilize the CCSP-2 antibody (Ab), and bovine serum albumin (BSA) used to prevent sensor surface fouling. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry (CV) were employed to analyze the electrochemical response to the binding of CCSP-2 antigen (Ag) and Ab. The changes in relative charge transfer resistance (ΔRct/Rcti) with varying concentrations of Ag were plotted and a calibration curve was established between ΔRct/Rcti and logarithm of Ag concentration to assess sensor’s sensitivity. The sensor demonstrated a linear response (R2 = 0.95) within the range of 10–100 ng/μL, plateauing after 100 ng/μL, with a detection limit of 0.71 ng/μL. Statistical analysis of specificity and selectivity studies showed significant differences in Ag detection compared to blank and nonspecific protein BSA, both with and without cell extracts. This immunosensor effectively detects the CRC biomarker CCSP-2 with high sensitivity and specificity. Integrating this sensor with other sensors for serum CRC biomarkers present a promising approach for developing diagnostic and prognostic tools for CRC.
期刊介绍：
ACS Measurement Science Au is an open access journal that publishes experimental computational or theoretical research in all areas of chemical measurement science. Short letters comprehensive articles reviews and perspectives are welcome on topics that report on any phase of analytical operations including sampling measurement and data analysis. This includes:Chemical Reactions and SelectivityChemometrics and Data ProcessingElectrochemistryElemental and Molecular CharacterizationImagingInstrumentationMass SpectrometryMicroscale and Nanoscale systemsOmics (Genomics Proteomics Metabonomics Metabolomics and Bioinformatics)Sensors and Sensing (Biosensors Chemical Sensors Gas Sensors Intracellular Sensors Single-Molecule Sensors Cell Chips Arrays Microfluidic Devices)SeparationsSpectroscopySurface analysisPapers dealing with established methods need to offer a significantly improved original application of the method.
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