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https://hdl.handle.net/10356/58910
Title: | Novel methods and screens for portein stability engineering | Authors: | Jose, Asial Ignacio | Keywords: | DRNTU::Science::Biological sciences::Biophysics DRNTU::Science::Biological sciences::Biochemistry DRNTU::Science::Biological sciences::Molecular biology |
Issue Date: | 2014 | Source: | Jose, A. I. (2014). Novel methods and screens for portein stability engineering. Doctoral thesis, Nanyang Technological University, Singapore. | Abstract: | Protein stability is often a limiting factor in the development of commercial proteins and biopharmaceuticals, as well as for biochemical and structural studies. In this thesis, I present different methods developed with the aim of improving protein stability, either by mutagenesis or by interaction with a protein partner.For protein stabilization through mutagenesis, a new high-throughput colony-based stability screen is described, which is a direct and biophysical read-out of intrinsic protein stability. As the method is generic and activity independent, it can easily be applied to a wide range of proteins. For protein stabilization through interaction with a protein partner, the use of phage display was explored. As an alternative and complement to phage display, I present the development of a new biophysical screen that monitors protein stabilization upon binding in cells and crude extracts, and its use in de novo binder generation using an autonomous VH domain as scaffold. | URI: | https://hdl.handle.net/10356/58910 | DOI: | 10.32657/10356/58910 | Schools: | School of Biological Sciences | Fulltext Permission: | open | Fulltext Availability: | With Fulltext |
Appears in Collections: | SBS Theses |
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File | Description | Size | Format | |
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ASIAL_PhD+thesis_final_2014.pdf | 19.09 MB | Adobe PDF | View/Open |
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