Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 29 for “"Yeast display"”.
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Applications of yeast display in synthetic biology with a focus on biological synthesis and recycling of plastics
Yeast surface display has emerged as a versatile tool in synthetic biology, particularly in protein engineering. This work sought to improve the flexibility and ease-of-use of yeast display for synthetic biologists, as well as applying the yeast display system in several contexts – with a …
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Engineering Antibodies to Control T Cell Function
In Chapter Five, a yeast display system was used to show that a T cell receptor ligand (scFv-KJ16) expressed on the surface of yeast was capable of inducing T cell activation. These findings suggest that the yeast display system can be used to discover novel polypeptides that regulate biological …
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Binding, Thermodynamic and Structural Studies of High-Affinity T Cell Receptor-Peptide MHC Interactions
… was used to show that scTCRs engineered in the yeast display system have similar stabilities as single chain antibody Fvs. The yeast display system was successful in determining a location for covlalent attachement of a fluorophore that did not disrupt the binding site. Labeled scTCRs were used …
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New protein engineering approaches for potentiating and studying antibody-based EGFR antagonism
… of a novel protein scaffold, called Sso7d, for yeast surface display-based libraries and selections. In the first part of this work I show that a combination of traditional yeast display techniques with simple but novel in silico approaches can be applied to derive a panel of Sso7d binders …
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Engineering T Cell Receptors to Examine the Role of Affinity and *Kinetics in T Helper Cell Function
… a patient with multiple sclerosis called 3A6. Yeast display of TCRs has previously relied on the use of an antibody specific for a properly folded TCR, here various strategies were used to generate stabilized 3A6 TCRs. Helpful strategies included: alteration of error-prone PCR dNTP …
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High-throughput tools for decoding T cell receptor specificity
… aim presents a new method that couples pMHC-displaying virus-like particles with yeast display, enabling efficient screening of millions of TCR variants against ~100 pMHCs at once. Yeast display is a powerful tool for studying TCR-antigen interactions but is constrained by its reliance on …
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Tool Development for Studying and Manipulating Peptide-MHC Interactions in a Globally-Representative Manner
… MHC alleles, utilizing data from high-throughput yeast display screens to train algorithms for antigen prediction. Next, we adapt the yeast display platform to screen user-defined libraries of peptides and apply the approach for optimizing peptides and profiling whole viral pathogens for MHC …
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Engineering T Cell Receptors for Potential Therapeutic Applications
… selections have involved incubating yeast cells with fluorescently labeled recombinant pepMHC ligand. Many pepMHC ligands are difficult to express and purify, so an alternative affinity selection strategy that obviates the need for this material would likewise facilitate affinity …
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Methylated DNA detection using a high-affinity methyl-CpG binding protein and photopolymerization-based amplification
… DNA methylation in biosensing applications. I displayed three of the core members MBD1, MBD2, and MBD4 on the surface of Saccharomyces cerevisiae yeast cells. Using the yeast display platform, I determined the equilibrium dissociation constant of human MBD2 (hMBD2) to be 5.9+/-1.3 nM for …
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Understanding the effects of higher order sequence features on peptide MHC binding
… representations to analyze data derived from yeast display platforms, which enable the collection of pMHC binding data for vast libraries of peptides. Methods for interpreting data from these platforms are still at an early stage, so in this thesis we also develop an approach for extracting …
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Structure, Function and Engineering of Peptide -Mhc Binding Receptors
… explored using various TCR mutants engineered by yeast display. Previous work in the lab showed that mutations in the CDR3 regions of the TCR could generate higher-affinity, peptide-specific TCR variants. In contrast, other CDRs (CDR1, and especially CDR2) of the TCR are typically located over the …
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Biologically inspired materials for electro-responsive coatings and the photo-oxidation of water
… as the nacre of abalone. Tools such as phage and yeast display libraries have enabled the combinatorial screening of peptides against a multitude of materials to which natural systems typically have no exposure. These techniques have yielded peptides that can bind and assemble technologically …
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Engineering and characterization of human single-chain T cell receptors
All nucleated cells display a sampling of their protein contents in the form of short 9-10 amino acid peptides bound to a product of the major histocompatibility complex (MHC) on the cell surface. This class of MHC-restricted antigens provides the broadest class of potential disease targets, as the …
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Determinants of antibody specificity
High throughput screening methods such as yeast surface display (YSD) are frequently able to isolate high affinity antibodies against clinical targets; however, the success of these candidates depends on selecting for both on-target binding and desirable biophysical characteristics. Development …
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Molecular Dissection of Bsc2: A Novel Negative Regulator of Triglyceride Lipolysis for a Lipid Droplet Subpopulation
… to specific LD subsets. Here, we show the yeast protein Bsc2 localizes to a specific subset of TG-containing LDs, and reveal it negatively regulates TG lipolysis. Mechanistically, Bsc2 LD targeting requires TG, and LD targeting is mediated by specific N-terminal hydrophobic regions (HRs) …
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Building a Genetic System in Yeast to Search for High Affinity Proteins in Sequence Space
… 2, I discuss the use of phenotypic selection for yeast-display protein binders, and test these systems on simple loop libraries. In Chapter 3, I construct a genetic system in yeast that can mutate a protein loop via homologous recombination, and test its recombination function. In Chapter 4, I …
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Development of lanthipeptide engineering strategies
… were explored to achieve this goal. First, a yeast display platform was used to select a precursor to a lanthipeptide that binds its biosynthetic enzymes more tightly, potentially enabling further biochemical characterization of a key substrate-enzyme interaction required for lanthipeptide …
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Recognition of hepatitis B epitopes by T cell receptors and T cell receptor-like antibodies
… engineering of T cell receptors expressed in a yeast display system. In view of the limitations imposed by the low affinity of soluble TCRs, protein engineering is necessary to improve the monomeric affinity (a process called affinity maturation). Conventional methods for screening …
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Molecular engineering of new protein labeling methodology based on rational design and in vitro evolution
… characterized a novel peptide substrate (called "yeast acceptor peptide" (yAP)) for yeast biotin ligase (yBL) that had been evolved by phage display. Assays performed in vitro and on the yeast surface showed that the yBL/yAP pair was orthogonal to the BirA/AP pair, allowing two-color labeling of …
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Investigation and application of substrate tolerant thioether forming enzymes
… for binding to UEV. To optimize XY3-3, two yeast display systems are investigated. Unfortunately, initial data suggested that the displayed peptide formed a disulfide bond which inhibited ProcM modification. Further optimization of yeast display systems or adaption of other display systems …
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