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Transient Expression of an Anthrax Decoy Fusion Protein in Glycoengineered Plant Cell Suspension Cultures
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| 자료유형 | 학위논문 |
|---|---|
| 서명/저자사항 | Transient Expression of an Anthrax Decoy Fusion Protein in Glycoengineered Plant Cell Suspension Cultures. |
| 개인저자 | Sukenik, Sara Conn. |
| 단체저자명 | University of California, Davis. Biomedical Engineering. |
| 발행사항 | [S.l.]: University of California, Davis., 2019. |
| 발행사항 | Ann Arbor: ProQuest Dissertations & Theses, 2019. |
| 형태사항 | 187 p. |
| 기본자료 저록 | Dissertations Abstracts International 81-06B. Dissertation Abstract International |
| ISBN | 9781392595695 |
| 학위논문주기 | Thesis (Ph.D.)--University of California, Davis, 2019. |
| 일반주기 |
Source: Dissertations Abstracts International, Volume: 81-06, Section: B.
Advisor: McDonald, Karen. |
| 이용제한사항 | This item must not be sold to any third party vendors. |
| 요약 | Conventional methods for recombinant protein production use stable transgenic cell lines, which require lengthy lead times for development before large-scale production. In an emergency situation, such as an infectious disease outbreak or bioterrorist attack, alternate methods are needed to enable large amounts of protein-based therapeutics to be produced in a shorter timeframe. In this dissertation, transient expression in plant cell suspension cultures was investigated as a recombinant protein production platform suitable for rapid response applications. Our system utilizes Agrobacterium tumefaciens, a plant pathogen that can be genetically engineered to deliver a gene of interest which is then expressed in the host plant cells. Different proteins can be produced using this method simply by co-culturing plant cells with different Agrobacterium constructs, which can be generated in as little time as two weeks. As a model protein, an anthrax receptor Fc fusion protein (CMG2-Fc) was used in process development studies. CMG2-Fc was successfully produced by co-culturing Agrobacterium and plant cell suspension cultures, with expression levels up to 9.8 關g/g plant cell fresh weight achieved after 7 days. To reduce the presence of plant-specific glycosylation patterns on the product, 棺-1,2-xylosyltransferase and 慣-1,3-fucosyltransferase knockdown Nicotiana benthamiana plant cell suspension cultures were used. Plant-specific glycan patterns were drastically reduced on CMG2-Fc produced in the glycoengineered N. benthamiana cell cultures compared to CMG2-Fc produced in wild type N. benthamiana plants. Various factors with potential to affect the interaction between Agrobacterium and plant cells were investigated to identify strategies to enhance recombinant protein production levels in the co-culture system. Finally, a techno-economic analysis of a large-scale co-culture process was performed, which indicated potential for cost-effective manufacturing and provided useful insights for future process development work. |
| 일반주제명 | Biomedical engineering. |
| 언어 | 영어 |
| 바로가기 | 
: 이 자료의 원문은 한국교육학술정보원에서 제공합니다. |
