A high-throughput screening method for engineering proteolytically stabilized growth factors
Abstract/Contents
- Abstract
- Growth factors are an important class of proteins that play a pivotal role in orchestrating many complex biological processes such as wound healing, tissue regeneration, vascularization, and tumor formation. Thus, there is immense interest in utilizing growth factors as protein therapeutics. However, although numerous recombinant growth factors have been developed as therapeutic candidates, only a few have been effective enough to receive clinical approval. This is due, in large part, to their generally poor stability. Therapeutic growth factors must remain active for an extended period to be efficacious. However, growth factors can become degraded upon exposure to enzymes known as proteases. A growth factor's susceptibility to proteases can be particularly important because these enzymes are especially active in areas relevant for disease treatment. Growth factors have previously been modified to improve their proteolytic stability, and this has been shown to enhance their biological activity. However, current methods of engineering growth factors for increased proteolytic stability are limited by low-throughput and the lack of generalizable strategies that can be used in a consistently successful manner. Thus, it can be a difficult and slow process to generate new growth factor variants with improved stability. In this work, we describe the development of a high-throughput screening method for engineering proteolytically stabilized growth factors. We validate the ability of the screen to differentiate between proteins of different proteolytic stabilities. Then, we utilize the screen to engineer a proteolytically stabilized variant of fibroblast growth factor 1 (FGF1). Finally, we characterize the ability of the engineered FGF1 protein to resist degradation, and show that this variant surprisingly antagonizes the FGFR cell signaling pathway, which may have therapeutic implications for the treatment of ocular neovascularization and tumor formation.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2018; ©2018 |
| Publication date | 2018; 2018 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Park, Sung Jin |
|---|---|
| Degree supervisor | Cochran, Jennifer R |
| Thesis advisor | Cochran, Jennifer R |
| Thesis advisor | Kim, Peter, 1958- |
| Thesis advisor | Lin, Michael Z |
| Degree committee member | Kim, Peter, 1958- |
| Degree committee member | Lin, Michael Z |
| Associated with | Stanford University, Department of Bioengineering. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Sung Jin Park. |
|---|---|
| Note | Submitted to the Department of Bioengineering. |
| Thesis | Thesis Ph.D. Stanford University 2018. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2018 by Sung Jin Park
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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