Beyond antibodies : high-throughput technologies for the development of synthetic affinity reagents with novel functions

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Abstract/Contents

Abstract
The accurate measurement of molecules is of crucial importance for a wide range of both basic and applied research applications. In particular, we need to measure the abundances and activities of biomolecules (proteins, carbohydrates, lipids, and nucleic acids) to understand their biological function which form the basis of life and diseases. To this end, we rely on affinity reagents, which are molecules that specifically bind to their target molecule. While antibodies are the current 'gold standard' of affinity reagents, there is a growing need for alternative reagents. Aptamers, often referred to as chemical antibodies, are short oligonucleotides that can bind to their targets with high specificity and affinity and can be engineered to perform novel functions that cannot be achieved with conventional antibodies. In this dissertation, I will discuss the development of three high-throughput technologies that enable the creation of synthetic DNA aptamers with novel functions. The first project describes a method to generate base-modified aptamers that can recognize protein glycoforms with exquisite specificity, which is challenging for antibodies. The second project describes the creation of a massively-parallel screen which enables the multiplexed selection of aptamers with extraordinary specificity that can distinguish metabolites with minimal difference in their chemical structure. Finally, I describe a screening technology for rapidly generating optically-responsive molecular switches from an existing aptamer which can be used directly as biosensors. As a whole, these studies provide valuable new tools that help address many of the challenges associated with the development of aptamers that are suitable for real-world applications.

Description

Type of resource text
Form electronic resource; remote; computer; online resource
Extent 1 online resource.
Place California
Place [Stanford, California]
Publisher [Stanford University]
Copyright date 2021; ©2021
Publication date 2021; 2021
Issuance monographic
Language English

Creators/Contributors

Author Yoshikawa, Alexander Masao
Degree supervisor Dunn, Alexander
Degree supervisor Soh, H. Tom
Thesis advisor Dunn, Alexander
Thesis advisor Soh, H. Tom
Thesis advisor Pitteri, Sharon
Degree committee member Pitteri, Sharon
Associated with Stanford University, Department of Chemical Engineering

Subjects

Genre Theses
Genre Text

Bibliographic information

Statement of responsibility Alexander Masao Yoshikawa.
Note Submitted to the Department of Chemical Engineering.
Thesis Thesis Ph.D. Stanford University 2021.
Location https://purl.stanford.edu/jy944hb5079

Access conditions

Copyright
© 2021 by Alexander Masao Yoshikawa
License
This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).

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