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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.
DOI: 10.32657/10356/58910
Fulltext Permission: open
Fulltext Availability: With Fulltext
Appears in Collections:SBS Theses

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