서브메뉴
검색
Plasma-Generated Hydroxyl Radicals for Epitope Mapping- [electronic resource]
![Plasma-Generated Hydroxyl Radicals for Epitope Mapping - [electronic resource]](/Users/Baul/Images/book.png)
Plasma-Generated Hydroxyl Radicals for Epitope Mapping- [electronic resource]
상세정보
- 자료유형
- 학위논문파일 국외
- 최종처리일시
- 20240214095838
- ISBN
- 9798380583343
- DDC
- 530
- 서명/저자
- Plasma-Generated Hydroxyl Radicals for Epitope Mapping - [electronic resource]
- 발행사항
- [S.l.]: : The University of Wisconsin - Madison., 2021
- 발행사항
- Ann Arbor : : ProQuest Dissertations & Theses,, 2021
- 형태사항
- 1 online resource(181 p.)
- 주기사항
- Source: Dissertations Abstracts International, Volume: 85-04, Section: B.
- 주기사항
- Advisor: Shohet, J. Leon.
- 학위논문주기
- Thesis (Ph.D.)--The University of Wisconsin - Madison, 2021.
- 사용제한주기
- This item must not be sold to any third party vendors.
- 초록/해제
- 요약A technology, Plasma Induced Modification of Biomolecules (PLIMB), has been developed for measuring three-dimensional (3-D) structural characteristics of proteins. It is used to perform hydroxyl-radical protein footprinting (HRPF) wherein a protein's solvent-accessible side chains are oxidized in solution using a hydroxyl-radical source and then analyzed with mass spectrometry. Although a technology like PLIMB can be used for many different applications, the unique characteristics of the technology and data it can produce may prove it to be uniquely qualified for the role of mapping the epitopes, or binding locations, of protein therapeutics to their target molecules. This is a crucial aspect of drug development and is important for drug optimization and intellectual-property filings. Although several proof-of-concept experiments demonstrating PLIMB's utility have been performed, additional research and development must be completed before PLIMB can be considered a viable option for epitope mapping. We hypothesize that by implementing technological improvements, processes, and methods, the epitopes for protein therapeutics can be mapped with PLIMB.This work first demonstrates the use of PLIMB for mapping the epitope of an antibody/antigen pair. Experimentation and data analysis using this procedure was effective, as several regions of the antigen were identified as the epitope region, which agree with previously published results.To address the recurring issue of variability in PLIMB data, a feedback-control system was developed, which uses UV-spectroscopy to measure hydroxyl radical generation in real time. Samples of the amino acid methionine were exposed to plasma and the amount of oxidation was measured with mass spectrometry. The results showed a reduction in the coefficient of variation between replicate samples from 15% to 6% when using the feedback control system, thus demonstrating its utility.To extract solvent accessibility measurements from PLIMB data, a procedure developed by other HRPF researchers was used. Samples of the protein myoglobin were exposed to various plasma-exposure times. Then a reactivity-normalized oxidation rate was calculated for several myoglobin amino acids. Results showed a linear correlation between reactivity-normalized oxidation rates and solvent-accessibility measurements generated from a published 3-D structure of myoglobin, and a root-mean-squared error in calculated solvent accessible surface areas of 24.98 A2 .
- 일반주제명
- Plasma physics.
- 일반주제명
- Biochemistry.
- 일반주제명
- Pharmaceutical sciences.
- 일반주제명
- Analytical chemistry.
- 일반주제명
- Electrical engineering.
- 키워드
- Myoglobin
- 키워드
- Epitope mapping
- 기타저자
- The University of Wisconsin - Madison Electrical Engineering
- 기본자료저록
- Dissertations Abstracts International. 85-04B.
- 기본자료저록
- Dissertation Abstract International
- 전자적 위치 및 접속
- 로그인 후 원문을 볼 수 있습니다.
MARC
008240612s2021 us |||||||||||||||c||eng d■001000016930935
■00520240214095838
■006m o d
■007cr#unu||||||||
■020 ▼a9798380583343
■035 ▼a(MiAaPQ)AAI28413059
■040 ▼aMiAaPQ▼cMiAaPQ
■0820 ▼a530
■1001 ▼aBenjamin, Daniel.
■24510▼aPlasma-Generated Hydroxyl Radicals for Epitope Mapping▼h[electronic resource]
■260 ▼a[S.l.]:▼bThe University of Wisconsin - Madison. ▼c2021
■260 1▼aAnn Arbor :▼bProQuest Dissertations & Theses, ▼c2021
■300 ▼a1 online resource(181 p.)
■500 ▼aSource: Dissertations Abstracts International, Volume: 85-04, Section: B.
■500 ▼aAdvisor: Shohet, J. Leon.
■5021 ▼aThesis (Ph.D.)--The University of Wisconsin - Madison, 2021.
■506 ▼aThis item must not be sold to any third party vendors.
■520 ▼aA technology, Plasma Induced Modification of Biomolecules (PLIMB), has been developed for measuring three-dimensional (3-D) structural characteristics of proteins. It is used to perform hydroxyl-radical protein footprinting (HRPF) wherein a protein's solvent-accessible side chains are oxidized in solution using a hydroxyl-radical source and then analyzed with mass spectrometry. Although a technology like PLIMB can be used for many different applications, the unique characteristics of the technology and data it can produce may prove it to be uniquely qualified for the role of mapping the epitopes, or binding locations, of protein therapeutics to their target molecules. This is a crucial aspect of drug development and is important for drug optimization and intellectual-property filings. Although several proof-of-concept experiments demonstrating PLIMB's utility have been performed, additional research and development must be completed before PLIMB can be considered a viable option for epitope mapping. We hypothesize that by implementing technological improvements, processes, and methods, the epitopes for protein therapeutics can be mapped with PLIMB.This work first demonstrates the use of PLIMB for mapping the epitope of an antibody/antigen pair. Experimentation and data analysis using this procedure was effective, as several regions of the antigen were identified as the epitope region, which agree with previously published results.To address the recurring issue of variability in PLIMB data, a feedback-control system was developed, which uses UV-spectroscopy to measure hydroxyl radical generation in real time. Samples of the amino acid methionine were exposed to plasma and the amount of oxidation was measured with mass spectrometry. The results showed a reduction in the coefficient of variation between replicate samples from 15% to 6% when using the feedback control system, thus demonstrating its utility.To extract solvent accessibility measurements from PLIMB data, a procedure developed by other HRPF researchers was used. Samples of the protein myoglobin were exposed to various plasma-exposure times. Then a reactivity-normalized oxidation rate was calculated for several myoglobin amino acids. Results showed a linear correlation between reactivity-normalized oxidation rates and solvent-accessibility measurements generated from a published 3-D structure of myoglobin, and a root-mean-squared error in calculated solvent accessible surface areas of 24.98 A2 .
■590 ▼aSchool code: 0262.
■650 4▼aPlasma physics.
■650 4▼aBiochemistry.
■650 4▼aPharmaceutical sciences.
■650 4▼aAnalytical chemistry.
■650 4▼aElectrical engineering.
■653 ▼aMyoglobin
■653 ▼aMass spectrometry
■653 ▼aEpitope mapping
■653 ▼aProtein therapeutics
■653 ▼aAmino acid methionine
■690 ▼a0759
■690 ▼a0487
■690 ▼a0572
■690 ▼a0544
■690 ▼a0486
■71020▼aThe University of Wisconsin - Madison▼bElectrical Engineering.
■7730 ▼tDissertations Abstracts International▼g85-04B.
■773 ▼tDissertation Abstract International
■790 ▼a0262
■791 ▼aPh.D.
■792 ▼a2021
■793 ▼aEnglish
■85640▼uhttp://www.riss.kr/pdu/ddodLink.do?id=T16930935▼nKERIS▼z이 자료의 원문은 한국교육학술정보원에서 제공합니다.
■980 ▼a202402▼f2024
