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Microscale Tools for Improved Analytical Sensitivity and Throughput in Single-Cell Immunoblotting- [electronic resource]
Microscale Tools for Improved Analytical Sensitivity and Throughput in Single-Cell Immunoblotting- [electronic resource]
상세정보
- 자료유형
- 학위논문파일 국외
- 최종처리일시
- 20240214095855
- ISBN
- 9798380621281
- DDC
- 610
- 서명/저자
- Microscale Tools for Improved Analytical Sensitivity and Throughput in Single-Cell Immunoblotting - [electronic resource]
- 발행사항
- [S.l.]: : University of California, Berkeley., 2021
- 발행사항
- Ann Arbor : : ProQuest Dissertations & Theses,, 2021
- 형태사항
- 1 online resource(144 p.)
- 주기사항
- Source: Dissertations Abstracts International, Volume: 85-04, Section: B.
- 주기사항
- Advisor: Herr, Amy E.
- 학위논문주기
- Thesis (Ph.D.)--University of California, Berkeley, 2021.
- 사용제한주기
- This item must not be sold to any third party vendors.
- 초록/해제
- 요약Proteins drive nearly all cellular processes, and direct quantitation of protein abundance from single-cells is essential to understanding heterogeneous cell states. Immunoassays are widely accepted tools for performing single-cell protein detection, but protein detection by immunoaffinity alone is insufficient for precision protein characterization, as proteins with similar binding kinetics can have different biological impacts, including in disease. To provide a cross-validation tool for protein characterization, electrophoretic cytometry immunoassays have been developed to characterize proteins by both immunoaffinity and molecular-mass through electrophoresis. Central to these assays performance is a multifunctional gel matrix that acts as a protein sieving matrix during electrophoresis and a protein scaffolding matrix during in-gel immunoblotting. In-gel immunoblotting of target proteins is widely accomplished by diffusively-driven immunoprobing, yet, this detection strategy suffers from reduced probe access to in-gel immobilized proteins via size-exclusion partitioning. Specifically, reduced probe delivery to the gel matrix in which target proteins are immobilized both (i) adversely impacts equilibrium immunocomplex formation and thus protein detection sensitivity and (ii) extends overall assay run time.In this dissertation, to improve the analytical detection capabilities and improve assay throughput in electrophoretic cytometry assays, we present methods to enhance immunoprobe delivery to hydrogel matrices, we introduce an assay design to improve throughput in single-cell immunoblotting, and we investigate reengineered sample handling designs for reduced protein losses before immobilization.Overall, we apply fundamentals in materials science, transport and reaction phenomena, and engineering design principles for the advancement of targeted protein detection assays. We see these advancements as contributing to the broader goal of improving our understanding of cell-state in healthy and disease conditions.
- 일반주제명
- Bioengineering.
- 일반주제명
- Physiology.
- 일반주제명
- Oncology.
- 일반주제명
- Immunology.
- 키워드
- Single-cells
- 키워드
- Immunoblotting
- 키워드
- Microscale tools
- 기타저자
- University of California, Berkeley Bioengineering
- 기본자료저록
- Dissertations Abstracts International. 85-04B.
- 기본자료저록
- Dissertation Abstract International
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
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■00520240214095855
■006m o d
■007cr#unu||||||||
■020 ▼a9798380621281
■035 ▼a(MiAaPQ)AAI28718055
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a610
■1001 ▼aMourdoukoutas, Antonios P.
■24510▼aMicroscale Tools for Improved Analytical Sensitivity and Throughput in Single-Cell Immunoblotting▼h[electronic resource]
■260 ▼a[S.l.]:▼bUniversity of California, Berkeley. ▼c2021
■260 1▼aAnn Arbor :▼bProQuest Dissertations & Theses, ▼c2021
■300 ▼a1 online resource(144 p.)
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-04, Section: B.
■500 ▼aAdvisor: Herr, Amy E.
■5021 ▼aThesis (Ph.D.)--University of California, Berkeley, 2021.
■506 ▼aThis item must not be sold to any third party vendors.
■520 ▼aProteins drive nearly all cellular processes, and direct quantitation of protein abundance from single-cells is essential to understanding heterogeneous cell states. Immunoassays are widely accepted tools for performing single-cell protein detection, but protein detection by immunoaffinity alone is insufficient for precision protein characterization, as proteins with similar binding kinetics can have different biological impacts, including in disease. To provide a cross-validation tool for protein characterization, electrophoretic cytometry immunoassays have been developed to characterize proteins by both immunoaffinity and molecular-mass through electrophoresis. Central to these assays performance is a multifunctional gel matrix that acts as a protein sieving matrix during electrophoresis and a protein scaffolding matrix during in-gel immunoblotting. In-gel immunoblotting of target proteins is widely accomplished by diffusively-driven immunoprobing, yet, this detection strategy suffers from reduced probe access to in-gel immobilized proteins via size-exclusion partitioning. Specifically, reduced probe delivery to the gel matrix in which target proteins are immobilized both (i) adversely impacts equilibrium immunocomplex formation and thus protein detection sensitivity and (ii) extends overall assay run time.In this dissertation, to improve the analytical detection capabilities and improve assay throughput in electrophoretic cytometry assays, we present methods to enhance immunoprobe delivery to hydrogel matrices, we introduce an assay design to improve throughput in single-cell immunoblotting, and we investigate reengineered sample handling designs for reduced protein losses before immobilization.Overall, we apply fundamentals in materials science, transport and reaction phenomena, and engineering design principles for the advancement of targeted protein detection assays. We see these advancements as contributing to the broader goal of improving our understanding of cell-state in healthy and disease conditions.
■590 ▼aSchool code: 0028.
■650 4▼aBioengineering.
■650 4▼aPhysiology.
■650 4▼aOncology.
■650 4▼aImmunology.
■653 ▼aCellular processes
■653 ▼aSingle-cells
■653 ▼aImmunoblotting
■653 ▼aMicroscale tools
■690 ▼a0202
■690 ▼a0992
■690 ▼a0982
■690 ▼a0719
■71020▼aUniversity of California, Berkeley▼bBioengineering.
■7730 ▼tDissertations Abstracts International▼g85-04B.
■773 ▼tDissertation Abstract International
■790 ▼a0028
■791 ▼aPh.D.
■792 ▼a2021
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T16931022▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
■980 ▼a202402▼f2024
