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Functionalized Nanoscale Materials f...
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Zhang, Youwen.
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Functionalized Nanoscale Materials for Protein Biomarker Detection.
紀錄類型:
書目-電子資源 : Monograph/item
正題名/作者:
Functionalized Nanoscale Materials for Protein Biomarker Detection./
作者:
Zhang, Youwen.
出版者:
Ann Arbor : ProQuest Dissertations & Theses, : 2020,
面頁冊數:
170 p.
附註:
Source: Dissertations Abstracts International, Volume: 82-09, Section: B.
Contained By:
Dissertations Abstracts International82-09B.
標題:
Analytical chemistry. -
電子資源:
https://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=27955128
ISBN:
9798582569978
Functionalized Nanoscale Materials for Protein Biomarker Detection.
Zhang, Youwen.
Functionalized Nanoscale Materials for Protein Biomarker Detection.
- Ann Arbor : ProQuest Dissertations & Theses, 2020 - 170 p.
Source: Dissertations Abstracts International, Volume: 82-09, Section: B.
Thesis (Ph.D.)--Illinois Institute of Technology, 2020.
This item must not be sold to any third party vendors.
Proteins are vital biomolecules in living organisms which function as the working element for many aspects of life. An abnormal expression of proteins or expression of unique proteins is often associated with certain disease. Accordingly, proteins have become valuable biomarkers for disease diagnosis and prognosis. So far, numerous methods have been developed for detections of protein biomarkers. However, most of them suffer from the lack of accuracy, sensitivity, and specificity for clinical diagnostic applications. With the rapid advancement in nanotechnology, functional nanoscale materials, which could overcome the biocompatibility and biological recognition ability, have been widely used to develop sensitive and selective biosensors.In this dissertation, two kinds of functionalized nanoscale materials-based sensing strategies are investigated for protein biomarker detection. One strategy takes advantages of graphene oxide (GO) and utilizes fluorescence resonance energy transfer (FRET) for fast and sensitive protease detection by covalent attaching fluorescently labeled protease substrate peptide to the GO surface. This type of GO-based fluorescence sensor is highly sensitive (with a detection limit of picomolar concentration) and selective (other structure similar proteases does not interfere with the target analyte detection). In addition, it could accurately analyze serum samples. With this strategy, we have successfully achieved the detection of the HIV-1 PR (HIV-1 protease, a significant biomarker for AIDS) and ADAMs (a disintegrin and metalloproteinases, a biomarker for human cancers). It could be visualized that this GO platform could be utilized to detect various proteases by only changing the peptide substrate and solution pH. In addition, by coupling multiple substrate peptides on the GO surface, we developed a multiplex GO sensing system for simultaneously profiling of the activities of a panel of MMPs/ADAMs. Under the assistance of joint entropy and programming, our sensor could identify up to 5 types of human cancers, and offers the potential to detect other cancer types by changing biomarkers.The other strategy is to utilize nanopore stochastic sensing to detect proteins, which involves measuring the ionic current modulation generated by analytes' electro-osmotic flow through a chemical functionalized nanoscale sized pore. As a sensitive and label-free technique, nanopores have been highly recognized as one of the emerging techniques to detect analytes at the single-molecule level. Unlike DNA molecules which are uniformly charged, proteins are an isotropically charged molecules, which have low translocation probability through a nanopore. Since the protein pore-based sensing system is not suitable as deployable tools for detection of proteins due to the size limitation and fragile nature of the biological membranes. In this project, we fabricated solid-state nanopores using PET membranes followed by chemical functionalization of their inner surfaces. The modified- PET nanopore was sensitive and could detect HIV-1 protease at picomolar concentration. More importantly, the modified-nanopore sensor was selective, and could differentiate the target protein from others such as Trypsin, BSA and HSA. Furthermore, the modified PET nanopore strategy developed in this work provide a general platform for exploring fundamental protein dynamics and rapid detection of proteins at the single-molecule level.
ISBN: 9798582569978Subjects--Topical Terms:
3168300
Analytical chemistry.
Subjects--Index Terms:
Biosensor
Functionalized Nanoscale Materials for Protein Biomarker Detection.
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Proteins are vital biomolecules in living organisms which function as the working element for many aspects of life. An abnormal expression of proteins or expression of unique proteins is often associated with certain disease. Accordingly, proteins have become valuable biomarkers for disease diagnosis and prognosis. So far, numerous methods have been developed for detections of protein biomarkers. However, most of them suffer from the lack of accuracy, sensitivity, and specificity for clinical diagnostic applications. With the rapid advancement in nanotechnology, functional nanoscale materials, which could overcome the biocompatibility and biological recognition ability, have been widely used to develop sensitive and selective biosensors.In this dissertation, two kinds of functionalized nanoscale materials-based sensing strategies are investigated for protein biomarker detection. One strategy takes advantages of graphene oxide (GO) and utilizes fluorescence resonance energy transfer (FRET) for fast and sensitive protease detection by covalent attaching fluorescently labeled protease substrate peptide to the GO surface. This type of GO-based fluorescence sensor is highly sensitive (with a detection limit of picomolar concentration) and selective (other structure similar proteases does not interfere with the target analyte detection). In addition, it could accurately analyze serum samples. With this strategy, we have successfully achieved the detection of the HIV-1 PR (HIV-1 protease, a significant biomarker for AIDS) and ADAMs (a disintegrin and metalloproteinases, a biomarker for human cancers). It could be visualized that this GO platform could be utilized to detect various proteases by only changing the peptide substrate and solution pH. In addition, by coupling multiple substrate peptides on the GO surface, we developed a multiplex GO sensing system for simultaneously profiling of the activities of a panel of MMPs/ADAMs. Under the assistance of joint entropy and programming, our sensor could identify up to 5 types of human cancers, and offers the potential to detect other cancer types by changing biomarkers.The other strategy is to utilize nanopore stochastic sensing to detect proteins, which involves measuring the ionic current modulation generated by analytes' electro-osmotic flow through a chemical functionalized nanoscale sized pore. As a sensitive and label-free technique, nanopores have been highly recognized as one of the emerging techniques to detect analytes at the single-molecule level. Unlike DNA molecules which are uniformly charged, proteins are an isotropically charged molecules, which have low translocation probability through a nanopore. Since the protein pore-based sensing system is not suitable as deployable tools for detection of proteins due to the size limitation and fragile nature of the biological membranes. In this project, we fabricated solid-state nanopores using PET membranes followed by chemical functionalization of their inner surfaces. The modified- PET nanopore was sensitive and could detect HIV-1 protease at picomolar concentration. More importantly, the modified-nanopore sensor was selective, and could differentiate the target protein from others such as Trypsin, BSA and HSA. Furthermore, the modified PET nanopore strategy developed in this work provide a general platform for exploring fundamental protein dynamics and rapid detection of proteins at the single-molecule level.
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