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Isolation, identification, and quantitation of biomolecules using high pressure liquid chromatography and mass spectrometry
Descriptive
TitleInfo
Title
Isolation, identification, and quantitation of biomolecules using high pressure liquid chromatography and mass spectrometry
Name
(type = personal)
NamePart
(type = family)
Greene
NamePart
(type = given)
Landon Ray
NamePart
(type = date)
1981-
DisplayForm
Landon Greene
Role
RoleTerm
(authority = RULIB)
author
Name
(type = personal)
NamePart
(type = family)
Lee
NamePart
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Jeehiun K
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Jeehiun K Lee
Affiliation
Advisory Committee
Role
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chair
Name
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NamePart
(type = family)
Krogh-Jesperson
NamePart
(type = given)
Karsten
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Karsten Krogh-Jesperson
Affiliation
Advisory Committee
Role
RoleTerm
(authority = RULIB)
internal member
Name
(type = personal)
NamePart
(type = family)
Hall
NamePart
(type = given)
Eugene S
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Eugene S Hall
Affiliation
Advisory Committee
Role
RoleTerm
(authority = RULIB)
internal member
Name
(type = personal)
NamePart
(type = family)
Buckley
NamePart
(type = given)
Brian
DisplayForm
Brian Buckley
Affiliation
Advisory Committee
Role
RoleTerm
(authority = RULIB)
outside member
Name
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NamePart
Rutgers University
Role
RoleTerm
(authority = RULIB)
degree grantor
Name
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NamePart
Graduate School - New Brunswick
Role
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(authority = RULIB)
school
TypeOfResource
Text
Genre
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theses
OriginInfo
DateCreated
(qualifier = exact)
2014
DateOther
(qualifier = exact);
(type = degree)
2014-10
CopyrightDate
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2014
Language
LanguageTerm
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(type = code)
eng
Abstract
(type = abstract)
The research performed in this dissertation involved isolation, identification, and quantitation of biomolecules using high pressure liquid chromatography and mass spectrometry. NanoRNAs, 2 – 4 nucleotide RNA oligomers, have been shown to prime transcription initiation in vivo. However, the particular oligonucleotides and absolute abundance could not be determined without the use of specific and sensitive analytical techniques. The goal of this research was to develop an analytical procedure to isolate, identify, and quantify nanoRNAs, in vivo, in Escherichia coli cells. A sequence of experiments was performed. A high performance pressure liquid chromatography (a.k.a. high pressure liquid chromatography) method was developed to isolate nanoRNA oligomers. This method was coupled with tandem mass spectrometry for detection. The combined HPLC-MS/MS method identified nanoRNAs in E. coli cell cultures. Experiments were performed to optimize the extraction of RNA oligomers from cell cultures to improve quantitation. An optimized extraction procedure and solid phase extraction method were developed which provided reproducible and quantitative analysis of nanoRNAs. The analysis of 5’, 3’-hydroxyl uridine-adenosine and 5’, 3’-hydroxyl uridine-adenosine-uridine was performed in E. coli cells. The results supported the qualitative hypothesis concerning the presence of these nanoRNAs. However, the absolute quantitation of nanoRNAs had significant error. Other research in this dissertation involved proteins. A novel monoclonal antibody, Das-1, was generated during ulcerative colitis research. This antibody has been used as a biomarker of various pre-cancerous and cancerous conditions of the gastrointestinal tract. However, the properties and structure of the specific antigen of mAb Das-1, a colonic epithelial protein (CEP), are unknown. The goal was to develop an isolation procedure to purify CEP. A three step procedure was developed. First, a size exclusion method was used to provide purification of CEP based on size. Second, a strong anion exchange method separated CEP based on overall surface charge. Finally, a hydrophobic interaction chromatography method provided purification based on hydrophobicity. The sequential purification of CEP using the three step procedure developed was tracked using several immunoassays. The results demonstrated significant purification of CEP.
Subject
(authority = RUETD)
Topic
Chemistry and Chemical Biology
Subject
(authority = ETD-LCSH)
Topic
Mass spectrometry
Subject
(authority = ETD-LCSH)
Topic
Liquid chromatography
Subject
(authority = ETD-LCSH)
Topic
Biomolecules--Analysis
RelatedItem
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TitleInfo
Title
Rutgers University Electronic Theses and Dissertations
Identifier
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ETD
Identifier
ETD_5754
PhysicalDescription
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electronic resource
InternetMediaType
application/pdf
InternetMediaType
text/xml
Extent
1 online resource (xii, 109 p. : ill.)
Note
(type = degree)
Ph.D.
Note
(type = bibliography)
Includes bibliographical references
Note
(type = statement of responsibility)
by Landon Ray Greene
RelatedItem
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TitleInfo
Title
Graduate School - New Brunswick Electronic Theses and Dissertations
Identifier
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rucore19991600001
Location
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(displayLabel = Rutgers, The State University of New Jersey)
NjNbRU
Identifier
(type = doi)
doi:10.7282/T38C9TQ4
Genre
(authority = ExL-Esploro)
ETD doctoral
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Rights
RightsDeclaration
(ID = rulibRdec0006)
The author owns the copyright to this work.
RightsHolder
(type = personal)
Name
FamilyName
Greene
GivenName
Landon
MiddleName
Ray
Role
Copyright Holder
RightsEvent
Type
Permission or license
DateTime
(encoding = w3cdtf);
(qualifier = exact);
(point = start)
2014-08-01 11:11:41
AssociatedEntity
Name
Landon Greene
Role
Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
AssociatedObject
Type
License
Name
Author Agreement License
Detail
I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
Copyright
Status
Copyright protected
Availability
Status
Open
Reason
Permission or license
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Technical
RULTechMD
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ContentModel
ETD
OperatingSystem
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windows xp
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