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Immunoproteomic identification of bovine pericardium xenoantigens

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dc.contributor.authorGriffiths, Leigh G., author
dc.contributor.authorOrton, E. Christopher, advisor
dc.contributor.authorReardon, Kenneth F., advisor
dc.date.accessioned2024-03-13T19:53:49Z
dc.date.available2024-03-13T19:53:49Z
dc.date.issued2008
dc.description.abstractBovine pericardium (BP) is an important biomaterial used in the production of gluteraldehyde-fixed heart valves and tissue engineering applications. The ability to perform proteomic analysis on BP is potentially useful for several reasons including investigation of immune rejection after implantation. The importance of humoral and cell mediated rejection responses towards such xenogeneic tissues are becoming increasingly apparent. I have applied a novel immunoproteomic approach to survey the antigenic determinants of BP. Proteomic analysis of fibrous tissues like BP is challenging due to their relative low cellularity and abundance of extracellular matrix. A variety of methods for tissue homogenization, protein extraction, and fractionation were investigated with the aim of producing high quality 2-DE gels for both water- and lipid-soluble BP proteins. MALDI-TOF/TOF MS protein identifications were performed to confirm bovine origin and appropriate subcellular fractionation of resolved proteins. Sixteen unique predominantly cytoplasmic bovine proteins were identified from the water-soluble gels. Twenty-two unique predominantly membrane bovine proteins were identified from the lipid-soluble gels. These results demonstrate that the final 2-DE protocol produced high quality proteomic data from BP for both cytoplasmic and membrane proteins. Duplicate 2-DE gels were used to generate western blots from both water- and lipid-soluble gels. Western blots were probed with pre- and post-exposure anti-BP rabbit serum, with detection of immune complexes limited to the IgG subtype. Western blots were compared to duplicate 2-DE gels and spots matched using Delta 2D image analysis software. Protein identifications of matched spots were performed using either MALDI-TOF/TOF MS or ESI MS/MS. This approach identified 31 putative antigens, capable of stimulating an IgG humoral rejection response. To the best of my knowledge, this study was the first to apply an immunoproteomic approach for identification of antigenic targets in xenotransplanted tissues. The results provide important information for understanding and possibly mitigating the immune response to fixed and unfixed BP xenografts.
dc.format.mediumborn digital
dc.format.mediumdoctoral dissertations
dc.identifierETDF_Griffiths_2008_3321280.pdf
dc.identifier.urihttps://hdl.handle.net/10217/237764
dc.languageEnglish
dc.language.isoeng
dc.publisherColorado State University. Libraries
dc.relation.ispartof2000-2019
dc.rightsCopyright and other restrictions may apply. User is responsible for compliance with all applicable laws. For information about copyright law, please see https://libguides.colostate.edu/copyright.
dc.rights.licensePer the terms of a contractual agreement, all use of this item is limited to the non-commercial use of Colorado State University and its authorized users.
dc.subjectantigen
dc.subjectbovine pericardium
dc.subjectimmunoproteomics
dc.subjectproteomics
dc.subjecttissue engineering
dc.subjectxenoantigens
dc.subjectanimal sciences
dc.subjectanimal diseases
dc.titleImmunoproteomic identification of bovine pericardium xenoantigens
dc.typeText
dcterms.rights.dplaThis Item is protected by copyright and/or related rights (https://rightsstatements.org/vocab/InC/1.0/). You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s).
thesis.degree.disciplineClinical Sciences
thesis.degree.grantorColorado State University
thesis.degree.levelDoctoral
thesis.degree.nameDoctor of Philosophy (Ph.D.)

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