MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR
Outbreaks of dengue hemorrhagic fever are caused by the dengue virus which belongs to the genus Flavivirus. This virus is carried and spread by the female Aedes-aegypti mosquito. Dengue virus has a single strand RNA (ssRNA) structure that has a size of 40-60 nm and has structural and non-structural...
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Teknik (Rekayasa, enjinering dan kegiatan berkaitan) Kusuma Dewi, Kariana MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR |
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Outbreaks of dengue hemorrhagic fever are caused by the dengue virus which belongs to the genus Flavivirus. This virus is carried and spread by the female Aedes-aegypti mosquito. Dengue virus has a single strand RNA (ssRNA) structure that has a size of 40-60 nm and has structural and non-structural proteins. In Indonesia, the incidence of dengue hemorrhagic fever is increasing every year, even Indonesia is one of the countries with the most dengue outbreaks in the world. One of the efforts to overcome this is by early detection of this virus, which generally uses the dengue virus NS1 antigen as the detected protein. In this study, an electrochemical biosensor based on MOF (Metal-organic Framework) HKUST-1 was developed which was modified with a triethanolamine modulator. HKUST-1 is a MOF material formed from metal Cu (Cupper) and organic ligand H3BTC (trimesic acid) and is one of the most studied MOF materials. The synthesis of this material uses the co-precipitation method with water as a solvent. Triethanolamine plays a role in controlling the pH of the reaction to provide a suitable environment for ion deprotonation and controlling the growth and structure of the resulting HKUST-1. The concentration of the addition of triethanolamine was varied, namely 20 wt%, 35 wt%, and 40 wt% and its effect was successfully observed through morphological changes that can be seen from the results of SEM (scanning electron microscopy). In addition, electrochemical performance was also observed to produce the best samples. Based on the results of cyclic voltammetry, the material with the best electrochemical performance was HKUST-1/TEOA 40% which was indicated by the highest peak oxidation current, so that this sample was further characterized to produce an optimal electrochemical immunosensor.
The DENV monoclonal antibody immobilization process was carried out using EDC-NHS coupling with 40% HKUST-1/TEOA matrix material. Electrochemical immunosensor performance was evaluated using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV) methods. All assays were carried out with the CorrTest electrochemical workstation instrument (Wuhan CorrTest Instruments Corp., Ltd China) on 3 mM K4[Fe(CN)6] electrolyte in 0.01 M PBS. The successful validation of the immobilization of each bioanalyte was observed by EIS assay, where each addition of antibody bioanalyte, BSA, and antigen resulted in a higher Rct (charge transfer resistance) value, indicating the success of the immobilization process in the matrix material. Electrochemical immunosensor response was observed from the DPV test, where the current response decreased proportionally with increasing concentration of NS-1 DENV-3 antigen and resulted in a detection limit of 0.932 pg/ml in a linear range of 0.001 – 10 ng/ml. This can be explained by the nature of the analyte which reduces or inhibits electron transfer in the surface-interface process of the material with the electrolyte because it closes the active site of Cu metal. Furthermore, reproducibility testing was carried out with 5 different GCEs but with the same preparation process showing fairly stable results. The selectivity was aimed to determine the possibility of the immunosensor in binding other analytes, experimentally the selectivity of the immunosensor was tested using 5 different analytes, namely bovine serum albumin (BSA), glucose, uric acid (UA), ascorbic acid (AA), urea, and NS-1 antigen. DENV-3 were respectively exposed on the surface of the immunosensor. Based on the analysis of the response results, a high selectivity value for the NS1 DENV-3 antigen was obtained, therefore, this immunosensor can be said to have good selectivity in detecting dengue virus serotype-3. Subsequent tests were stability and reusability within a 3-week storage range, indicating that the immunosensor was stable for 3 weeks. Furthermore, to determine the response of the immunosensor in clinical testing, immunosensor testing was carried out in human serum diluted 100x with 0.01 M PBS solution showing optimal results, so this immunosensor has the potential for further development as a point-of-care device (POC) electrochemical immunosensor for early detection of NS1 DENV-3 antigen. |
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Theses |
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Kusuma Dewi, Kariana |
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Kusuma Dewi, Kariana |
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Kusuma Dewi, Kariana |
title |
MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR |
title_short |
MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR |
title_full |
MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR |
title_fullStr |
MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR |
title_full_unstemmed |
MODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR |
title_sort |
modification of hkust-1 (cubtc) by addition triethanolamine for early detection application of dengue virus serorype-3 based on electrochemical immunosensor |
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id-itb.:799072024-01-16T14:16:58ZMODIFICATION OF HKUST-1 (CUBTC) BY ADDITION TRIETHANOLAMINE FOR EARLY DETECTION APPLICATION OF DENGUE VIRUS SERORYPE-3 BASED ON ELECTROCHEMICAL IMMUNOSENSOR Kusuma Dewi, Kariana Teknik (Rekayasa, enjinering dan kegiatan berkaitan) Indonesia Theses electrochemical immunosensor, HKUST-1 modification, triethanolamine, NS1 DENV-3 antigen INSTITUT TEKNOLOGI BANDUNG https://digilib.itb.ac.id/gdl/view/79907 Outbreaks of dengue hemorrhagic fever are caused by the dengue virus which belongs to the genus Flavivirus. This virus is carried and spread by the female Aedes-aegypti mosquito. Dengue virus has a single strand RNA (ssRNA) structure that has a size of 40-60 nm and has structural and non-structural proteins. In Indonesia, the incidence of dengue hemorrhagic fever is increasing every year, even Indonesia is one of the countries with the most dengue outbreaks in the world. One of the efforts to overcome this is by early detection of this virus, which generally uses the dengue virus NS1 antigen as the detected protein. In this study, an electrochemical biosensor based on MOF (Metal-organic Framework) HKUST-1 was developed which was modified with a triethanolamine modulator. HKUST-1 is a MOF material formed from metal Cu (Cupper) and organic ligand H3BTC (trimesic acid) and is one of the most studied MOF materials. The synthesis of this material uses the co-precipitation method with water as a solvent. Triethanolamine plays a role in controlling the pH of the reaction to provide a suitable environment for ion deprotonation and controlling the growth and structure of the resulting HKUST-1. The concentration of the addition of triethanolamine was varied, namely 20 wt%, 35 wt%, and 40 wt% and its effect was successfully observed through morphological changes that can be seen from the results of SEM (scanning electron microscopy). In addition, electrochemical performance was also observed to produce the best samples. Based on the results of cyclic voltammetry, the material with the best electrochemical performance was HKUST-1/TEOA 40% which was indicated by the highest peak oxidation current, so that this sample was further characterized to produce an optimal electrochemical immunosensor. The DENV monoclonal antibody immobilization process was carried out using EDC-NHS coupling with 40% HKUST-1/TEOA matrix material. Electrochemical immunosensor performance was evaluated using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV) methods. All assays were carried out with the CorrTest electrochemical workstation instrument (Wuhan CorrTest Instruments Corp., Ltd China) on 3 mM K4[Fe(CN)6] electrolyte in 0.01 M PBS. The successful validation of the immobilization of each bioanalyte was observed by EIS assay, where each addition of antibody bioanalyte, BSA, and antigen resulted in a higher Rct (charge transfer resistance) value, indicating the success of the immobilization process in the matrix material. Electrochemical immunosensor response was observed from the DPV test, where the current response decreased proportionally with increasing concentration of NS-1 DENV-3 antigen and resulted in a detection limit of 0.932 pg/ml in a linear range of 0.001 – 10 ng/ml. This can be explained by the nature of the analyte which reduces or inhibits electron transfer in the surface-interface process of the material with the electrolyte because it closes the active site of Cu metal. Furthermore, reproducibility testing was carried out with 5 different GCEs but with the same preparation process showing fairly stable results. The selectivity was aimed to determine the possibility of the immunosensor in binding other analytes, experimentally the selectivity of the immunosensor was tested using 5 different analytes, namely bovine serum albumin (BSA), glucose, uric acid (UA), ascorbic acid (AA), urea, and NS-1 antigen. DENV-3 were respectively exposed on the surface of the immunosensor. Based on the analysis of the response results, a high selectivity value for the NS1 DENV-3 antigen was obtained, therefore, this immunosensor can be said to have good selectivity in detecting dengue virus serotype-3. Subsequent tests were stability and reusability within a 3-week storage range, indicating that the immunosensor was stable for 3 weeks. Furthermore, to determine the response of the immunosensor in clinical testing, immunosensor testing was carried out in human serum diluted 100x with 0.01 M PBS solution showing optimal results, so this immunosensor has the potential for further development as a point-of-care device (POC) electrochemical immunosensor for early detection of NS1 DENV-3 antigen. text |