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Development and Optimization of High-Throughput Cell-Free Platforms for Biosensor Engineering
Development and Optimization of High-Throughput Cell-Free Platforms for Biosensor Engineering
Detailed Information
- 자료유형
- 학위논문 서양
- 최종처리일시
- 20250211152730
- ISBN
- 9798384016359
- DDC
- 660
- 저자명
- Ekas, Holly Mei.
- 서명/저자
- Development and Optimization of High-Throughput Cell-Free Platforms for Biosensor Engineering
- 발행사항
- [Sl] : Northwestern University, 2024
- 발행사항
- Ann Arbor : ProQuest Dissertations & Theses, 2024
- 형태사항
- 127 p
- 주기사항
- Source: Dissertations Abstracts International, Volume: 86-02, Section: B.
- 주기사항
- Advisor: Jewett, Michael C.
- 학위논문주기
- Thesis (Ph.D.)--Northwestern University, 2024.
- 초록/해제
- 요약At our current trajectory environmental catastrophes will become mainstays of our global society by 2050. With failing infrastructure and unhindered agricultural runoff, the long-term ramifications of consuming contaminated water will be felt by current and future generations alike. Current analytical chemistry methods provide accurate and sensitive techniques for detecting even trace amounts of contaminants in water samples using highly advanced equipment harbored in centralized laboratories. This detection pipeline is slow, expensive, and inaccessible, especially in resource limited settings where diagnostics are most crucial. Biological systems have naturally developed sense and respond mechanisms that can be re-engineered in chassis strains to enable whole-cell in vivo biosensors, circumventing instrumentation requirements by producing visible signal in response to analyte. However, biocontainment and instability concerns prevent them from seeing practical application at the point-of-use. Cell-free biosensors offer an in vitro platform for field-deployable diagnostics that is cheaper, safer, shelf-stable, and more easily engineered. I outline the field of biosensor technologies and current shortcomings for practical use, highlighting the promising utility of transcription factor biosensors for the detection of water contaminants. In this dissertation, I describe my work to develop a high-throughput screening platform that can rapidly assemble and assess thousands of unique cell-free biosensing reactions in parallel. I develop a quantitative assay development workflow that can be tailored to any transcription factor. I apply this platform towards engineering transcription factors relevant to water quality diagnostics: MerR (mercury-sensing), CadR (cadmium-sensing), and PbrR (lead-sensing). I demonstrate the efficacy of this platform by engineering PbrR to detect lead at legal limit set by the Environmental Protection Agency. I then describe future works that could expand upon this foundation, and preliminary data demonstrating the utility of interfacing this platform with machine learning. I also propose novel high-throughput pipelines. Altogether, the innovations in this work will enable biologists to rapidly generate critical diagnostics to combat this global health crisis.
- 일반주제명
- Chemical engineering
- 일반주제명
- Biomedical engineering
- 일반주제명
- Medical imaging
- 키워드
- Global society
- 키워드
- Biosensors
- 키워드
- Cadmium-sensing
- 키워드
- Machine learning
- 기타저자
- Northwestern University Chemical and Biological Engineering
- 기본자료저록
- Dissertations Abstracts International. 86-02B.
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
008250123s2024 us c eng d■001000017163602
■00520250211152730
■006m o d
■007cr#unu||||||||
■020 ▼a9798384016359
■035 ▼a(MiAaPQ)AAI31490680
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a660
■1001 ▼aEkas, Holly Mei.
■24510▼aDevelopment and Optimization of High-Throughput Cell-Free Platforms for Biosensor Engineering
■260 ▼a[Sl]▼bNorthwestern University▼c2024
■260 1▼aAnn Arbor▼bProQuest Dissertations & Theses▼c2024
■300 ▼a127 p
■500 ▼aSource: Dissertations Abstracts International, Volume: 86-02, Section: B.
■500 ▼aAdvisor: Jewett, Michael C.
■5021 ▼aThesis (Ph.D.)--Northwestern University, 2024.
■520 ▼aAt our current trajectory environmental catastrophes will become mainstays of our global society by 2050. With failing infrastructure and unhindered agricultural runoff, the long-term ramifications of consuming contaminated water will be felt by current and future generations alike. Current analytical chemistry methods provide accurate and sensitive techniques for detecting even trace amounts of contaminants in water samples using highly advanced equipment harbored in centralized laboratories. This detection pipeline is slow, expensive, and inaccessible, especially in resource limited settings where diagnostics are most crucial. Biological systems have naturally developed sense and respond mechanisms that can be re-engineered in chassis strains to enable whole-cell in vivo biosensors, circumventing instrumentation requirements by producing visible signal in response to analyte. However, biocontainment and instability concerns prevent them from seeing practical application at the point-of-use. Cell-free biosensors offer an in vitro platform for field-deployable diagnostics that is cheaper, safer, shelf-stable, and more easily engineered. I outline the field of biosensor technologies and current shortcomings for practical use, highlighting the promising utility of transcription factor biosensors for the detection of water contaminants. In this dissertation, I describe my work to develop a high-throughput screening platform that can rapidly assemble and assess thousands of unique cell-free biosensing reactions in parallel. I develop a quantitative assay development workflow that can be tailored to any transcription factor. I apply this platform towards engineering transcription factors relevant to water quality diagnostics: MerR (mercury-sensing), CadR (cadmium-sensing), and PbrR (lead-sensing). I demonstrate the efficacy of this platform by engineering PbrR to detect lead at legal limit set by the Environmental Protection Agency. I then describe future works that could expand upon this foundation, and preliminary data demonstrating the utility of interfacing this platform with machine learning. I also propose novel high-throughput pipelines. Altogether, the innovations in this work will enable biologists to rapidly generate critical diagnostics to combat this global health crisis.
■590 ▼aSchool code: 0163.
■650 4▼aChemical engineering
■650 4▼aBiomedical engineering
■650 4▼aMedical imaging
■653 ▼aGlobal society
■653 ▼aBiosensors
■653 ▼aEnvironmental catastrophes
■653 ▼aCadmium-sensing
■653 ▼aMachine learning
■690 ▼a0542
■690 ▼a0541
■690 ▼a0574
■71020▼aNorthwestern University▼bChemical and Biological Engineering.
■7730 ▼tDissertations Abstracts International▼g86-02B.
■790 ▼a0163
■791 ▼aPh.D.
■792 ▼a2024
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T17163602▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
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