Thesis
The characterization of calmodulin-integrated polyethylene glycol diacrylate hydrogels using the model protein GB1
Washington State University
Master of Science (MS), Washington State University
2016
Handle:
https://hdl.handle.net/2376/103040
Abstract
Polyethylene glycol diacrylate (PEGDA) conjugated protein hydrogels are a promising new biomaterial for use in protein preservation and delivery. We have characterized the ability of a PEGDA-calmodulin hydrogel conjugate to bind, preserve and release an M13-tagged model protein, the B1 domain from streptococcus protein G (GB1). We used solution-state nuclear magnetic resonance (NMR) spectroscopy to examine the response of the hydrogel to thermal and desiccation stress. Our data shows that the PEGDA-calmodulin hydrogel can bind the M13-tagged GB1 without significantly disturbing the structure of the model protein. Addition of the calcium chelator, EGTA, disrupts the interaction between calmodulin and the M13 tag, resulting in the release of the protein from the hydrogel. We found that the PEGDA-calmodulin hydrogel provided little thermal stabilization of the protein when compared to the complex in solution. In addition, the hydrogel retained the model protein through desiccation and re-hydration, as well as its ability to release the protein upon addition of EGTA. Finally, when IgG was soaked into the gels, GB1 bound IgG and both IgG and GB1 were released with the addition of EGTA. These results demonstrate that calmodulin PEGDA hydrogels are a viable protein immobilization tool capable of preserving proteins and protein function.
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Details
- Title
- The characterization of calmodulin-integrated polyethylene glycol diacrylate hydrogels using the model protein GB1
- Creators
- Carrie Lynn Marean-Reardon
- Contributors
- Allan S. Felsot (Degree Supervisor)
- Awarding Institution
- Washington State University
- Academic Unit
- Environment, School of the (CAHNRS)
- Theses and Dissertations
- Master of Science (MS), Washington State University
- Publisher
- Washington State University; [Pullman, Washington] :
- Identifiers
- 99900525142901842
- Language
- English
- Resource Type
- Thesis