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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 Engineer...
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  
Environmental catastrophes
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

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■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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