CELL ENGINEERING AND CULTURE MEDIA MODIFICATION STUDIES TO IMPROVE THE CELL CULTURE PERFORMANCE
| dc.contributor.advisor | Betenbaugh, Michael J | |
| dc.contributor.committeeMember | Ostermeier, Marc | |
| dc.creator | Dhara, Venkata Gayatri | |
| dc.date.accessioned | 2019-04-15T03:26:24Z | |
| dc.date.created | 2017-08 | |
| dc.date.issued | 2017-07-18 | |
| dc.date.submitted | August 2017 | |
| dc.date.updated | 2019-04-15T03:26:24Z | |
| dc.description.abstract | Mammalian cell cultures have become a universal path for producing therapeutic proteins, monoclonal antibodies in particular, in biopharmaceutical industries. There is a constant attempt to improve the productivity of the cell cultures through research in the industry and educational institutions. The productivity of the cells can be improved through various techniques. The use of additives is the most economical option to pursue for improving the productivity of the cells. The additives, hydrolysates, were used in the cell culture at a final concentration of 0.4% and we have seen a two to threefold increase in the productivity of the cells. While soy hydrolysate doubled the productivity along with the cell growth, cotton hydrolysates have increased it three times when compared to the control flasks not supplemented with any hydrolysates. Five different lots of cotton hydrolysates were tested simultaneously as media supplements for a lot variability study. The cotton lots 100NTCR and 100PCHO performed the best among the five lots tested in terms of both cell growth and IgG productivity. Both these lots were selected to perform proteomics analysis on the hydrolysate samples to understand the mode of action of these hydrolysates when added to the cell cultures. Essential amino acids’ biosynthetic pathways were to be engineered into mammalian cells. In particular, the research described in the thesis targeted branched chain amino acids- Leucine, Isoleucine and Valine. All the genes were cloned into the mammalian expression vector pBUD4.1 and later transfected into CHO-K1 and HEK 293T cell lines and their expression was tested using a western blot. Five out of the nine genes to be expressed, have worked. This opens up new avenues to explore the machinery in the mammalian cell and lead to the synthesis of a minimal cell. | |
| dc.format.mimetype | application/pdf | |
| dc.identifier.uri | http://jhir.library.jhu.edu/handle/1774.2/60428 | |
| dc.language.iso | en_US | |
| dc.publisher | Johns Hopkins University | |
| dc.publisher.country | USA | |
| dc.subject | Mammalian cell engineering, Mammalian cell culture, amino acids, hydrolysates | |
| dc.title | CELL ENGINEERING AND CULTURE MEDIA MODIFICATION STUDIES TO IMPROVE THE CELL CULTURE PERFORMANCE | |
| dc.type | Thesis | |
| dc.type.material | text | |
| local.embargo.lift | 2021-08-01 | |
| local.embargo.terms | 2021-08-01 | |
| thesis.degree.department | Chemical and Biomolecular Engineering | |
| thesis.degree.discipline | Biotechnology | |
| thesis.degree.grantor | Johns Hopkins University | |
| thesis.degree.grantor | Whiting School of Engineering | |
| thesis.degree.level | Masters | |
| thesis.degree.name | M.S.E. |