STUDY AND CHARACTERIZATION OF DUAL-FUNCTION AFFINITY CHROMATOGRAPHIC ADSORBENTS HAVING SIZE EXCLUSION AND ADSORPTION PROPERTIES TO ISOLATE, PURIFY AND RECOVER SMALL BIOMOLECULES FROM COMPLEX BIOLOGICAL MIXTURES

Persistent Link:
http://hdl.handle.net/10150/195896
Title:
STUDY AND CHARACTERIZATION OF DUAL-FUNCTION AFFINITY CHROMATOGRAPHIC ADSORBENTS HAVING SIZE EXCLUSION AND ADSORPTION PROPERTIES TO ISOLATE, PURIFY AND RECOVER SMALL BIOMOLECULES FROM COMPLEX BIOLOGICAL MIXTURES
Author:
Gonzalez Ortega, Omar
Issue Date:
2010
Publisher:
The University of Arizona.
Rights:
Copyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.
Abstract:
In this work, the main emphasis of the research concerns the development of isolation and purification methods of biomolecules from biological fluids. Several separation techniques were incorporated in chromatographic gels to obtain multifunctional hybrid chromatographic separation media for proteins, peptides and amino acid isolation and purification.In the first part of the research, several chelating agents were synthesized and their effectiveness to purify immunoglobulins using Immobilized Metal Affinity Chromatography (IMAC) was investigated. Ethylenediamine triacetic acid (TED) with immobilized copper resulted in the most effective in terms of purification and protein capacities.The next part of the work involved the development of hybrid chromatographic media that combines protein specific adsorption with sharp controlled size access permeation. This was accomplished by incorporating two types of ligand derivatives, one that permits the permeation of only certain molecular size range compounds, and a second one that specifically binds target biomolecules among the compounds of that specific molecular size range. Hybrid systems included binding ligands for Immobilized Metal Affinity Chromatography (IMAC), Ion Exchange Chromatography (IEX) and Hydrophobic Interaction Chromatography (HIC) combined with a controlled access polymer at different densities such as polyethylene glycol (PEG) and dextran derivatives. In general, low grafting density of high molecular weight PEG was found to be as effective as high grafting density of low molecular weight PEG in the rejecting properties of the semi-permeable synthesized media.Theoretical and experimental batch adsorption studies were also performed with the hybrid media and a mathematical model was developed to study the uptake of proteins under specific conditions of controlled permeation.In the last stage of this work, chelating surfactants were synthesized and used as reversible affinity ligands on reversed phase adsorbents for protein separations.One of the main accomplishments of this research was the development of separation media for small molecular size compounds from larger molecules and from complex biological systems. Applications of special interest will include the isolation and purification of solutes, such as metal ions, toxins, drugs, biomolecules, including proteins, biotoxins, nucleic acids, peptides, hormones, and biomarkers from biological fluids (such as human serum, urine, etc.) and from aqueous solutions.
Type:
text; Electronic Dissertation
Keywords:
access; bioseparations; media; proteins; restricted; serum
Degree Name:
Ph.D.
Degree Level:
doctoral
Degree Program:
Chemical Engineering; Graduate College
Degree Grantor:
University of Arizona
Advisor:
Guzman, Roberto
Committee Chair:
Guzman, Roberto

Full metadata record

DC FieldValue Language
dc.language.isoenen_US
dc.titleSTUDY AND CHARACTERIZATION OF DUAL-FUNCTION AFFINITY CHROMATOGRAPHIC ADSORBENTS HAVING SIZE EXCLUSION AND ADSORPTION PROPERTIES TO ISOLATE, PURIFY AND RECOVER SMALL BIOMOLECULES FROM COMPLEX BIOLOGICAL MIXTURESen_US
dc.creatorGonzalez Ortega, Omaren_US
dc.contributor.authorGonzalez Ortega, Omaren_US
dc.date.issued2010en_US
dc.publisherThe University of Arizona.en_US
dc.rightsCopyright © is held by the author. Digital access to this material is made possible by the University Libraries, University of Arizona. Further transmission, reproduction or presentation (such as public display or performance) of protected items is prohibited except with permission of the author.en_US
dc.description.abstractIn this work, the main emphasis of the research concerns the development of isolation and purification methods of biomolecules from biological fluids. Several separation techniques were incorporated in chromatographic gels to obtain multifunctional hybrid chromatographic separation media for proteins, peptides and amino acid isolation and purification.In the first part of the research, several chelating agents were synthesized and their effectiveness to purify immunoglobulins using Immobilized Metal Affinity Chromatography (IMAC) was investigated. Ethylenediamine triacetic acid (TED) with immobilized copper resulted in the most effective in terms of purification and protein capacities.The next part of the work involved the development of hybrid chromatographic media that combines protein specific adsorption with sharp controlled size access permeation. This was accomplished by incorporating two types of ligand derivatives, one that permits the permeation of only certain molecular size range compounds, and a second one that specifically binds target biomolecules among the compounds of that specific molecular size range. Hybrid systems included binding ligands for Immobilized Metal Affinity Chromatography (IMAC), Ion Exchange Chromatography (IEX) and Hydrophobic Interaction Chromatography (HIC) combined with a controlled access polymer at different densities such as polyethylene glycol (PEG) and dextran derivatives. In general, low grafting density of high molecular weight PEG was found to be as effective as high grafting density of low molecular weight PEG in the rejecting properties of the semi-permeable synthesized media.Theoretical and experimental batch adsorption studies were also performed with the hybrid media and a mathematical model was developed to study the uptake of proteins under specific conditions of controlled permeation.In the last stage of this work, chelating surfactants were synthesized and used as reversible affinity ligands on reversed phase adsorbents for protein separations.One of the main accomplishments of this research was the development of separation media for small molecular size compounds from larger molecules and from complex biological systems. Applications of special interest will include the isolation and purification of solutes, such as metal ions, toxins, drugs, biomolecules, including proteins, biotoxins, nucleic acids, peptides, hormones, and biomarkers from biological fluids (such as human serum, urine, etc.) and from aqueous solutions.en_US
dc.typetexten_US
dc.typeElectronic Dissertationen_US
dc.subjectaccessen_US
dc.subjectbioseparationsen_US
dc.subjectmediaen_US
dc.subjectproteinsen_US
dc.subjectrestricteden_US
dc.subjectserumen_US
thesis.degree.namePh.D.en_US
thesis.degree.leveldoctoralen_US
thesis.degree.disciplineChemical Engineeringen_US
thesis.degree.disciplineGraduate Collegeen_US
thesis.degree.grantorUniversity of Arizonaen_US
dc.contributor.advisorGuzman, Robertoen_US
dc.contributor.chairGuzman, Robertoen_US
dc.contributor.committeememberBaygents, Jimen_US
dc.contributor.committeememberSaez, Eduardoen_US
dc.identifier.proquest11180en_US
dc.identifier.oclc752261034en_US
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