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020 ▼a 9781392595695
035 ▼a (MiAaPQ)AAI13901381
040 ▼a MiAaPQ ▼c MiAaPQ ▼d 247004
0820 ▼a 610
1001 ▼a Sukenik, Sara Conn.
24510 ▼a Transient Expression of an Anthrax Decoy Fusion Protein in Glycoengineered Plant Cell Suspension Cultures.
260 ▼a [S.l.]: ▼b University of California, Davis., ▼c 2019.
260 1 ▼a Ann Arbor: ▼b ProQuest Dissertations & Theses, ▼c 2019.
300 ▼a 187 p.
500 ▼a Source: Dissertations Abstracts International, Volume: 81-06, Section: B.
500 ▼a Advisor: McDonald, Karen.
5021 ▼a Thesis (Ph.D.)--University of California, Davis, 2019.
506 ▼a This item must not be sold to any third party vendors.
520 ▼a 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.
590 ▼a School code: 0029.
650 4 ▼a Biomedical engineering.
690 ▼a 0541
71020 ▼a University of California, Davis. ▼b Biomedical Engineering.
7730 ▼t Dissertations Abstracts International ▼g 81-06B.
773 ▼t Dissertation Abstract International
790 ▼a 0029
791 ▼a Ph.D.
792 ▼a 2019
793 ▼a English
85640 ▼u http://www.riss.kr/pdu/ddodLink.do?id=T15492289 ▼n KERIS ▼z 이 자료의 원문은 한국교육학술정보원에서 제공합니다.
980 ▼a 202002 ▼f 2020
990 ▼a ***1008102
991 ▼a E-BOOK