The Production of Recombinant Immunoglobulin Superfamily Proteins in the Yeast Saccharomyces Cerevisiae
Shusta, Eric Victor
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https://hdl.handle.net/2142/82475
Description
Title
The Production of Recombinant Immunoglobulin Superfamily Proteins in the Yeast Saccharomyces Cerevisiae
Author(s)
Shusta, Eric Victor
Issue Date
1999
Doctoral Committee Chair(s)
Wittrup, K. Dane
Department of Study
Chemical Engineering
Discipline
Chemical Engineering
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Language
eng
Abstract
Although scFvs and scTCRs are structurally homologous, they have very distinct expression and stability behaviors. In fact, in order to develop an efficient scTCR production strategy like that developed for scFvs, it was necessary to alter the characteristics of the protein itself to improve its processing and stability characteristics. These properties were improved using a directed evolution approach and yeast surface display. Evolution resulted in a super-scTCR scaffold with optimized secretion, solubility, and stability properties. This scaffold will, for the first time, enable directed evolution of the binding properties of the scTCR for its peptide/MHC ligand. The results described in this thesis illustrate the power of combining genetic, cellular, and protein engineering tools for improving the secretion and stability characteristics of a target protein.
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