Advancements in the development of quantitative molecular assays
Abstract/Contents
- Abstract
- The first published implementation of a binding assay was an immunoassay for insulin detection in 1960. Over the subsequent 60 years, analytical technologies based on binding assays have evolved substantially. Molecular detection platforms based on enzyme-linked immunosorbent assays (ELISAs)—as well as newer commercial technologies, such as Luminex and NanoString—are core components of today's diagnostic and research armamentaria. There are two main challenges in the development of binding assays. First, binding alone does not generate an observable signal. The target and/or affinity reagent must be directly coupled to a moiety that generates a signal upon binding. Myriad immunosensors have been developed that couple the interaction between an antibody and its antigen to an observable output based on an electronic, optical, or mechanical signal. The second challenge is that binding assays are governed by the principles of chemical equilibrium. Binding assays generate signal through concentration-dependent shifts in equilibrium that result from the interaction of the target molecule with an affinity reagent. Convention affixes the range of measurable target concentrations to an 81-fold dynamic range centered around the dissociation constant of an affinity reagent to its target. In this dissertation, I introduce three major advances to the development of binding assays: (i) I demonstrate that high-throughput DNA sequencing can be used as a readout mechanism that is in many ways superior to optical and electrical readouts, (ii) I describe novel methodologies of modulating detection ranges without changing the actual physicochemical interactions between target and affinity reagent, and (iii) I challenge the conventional wisdom regarding binding assays and introduce a new conceptual framework for developing modern analytical technologies. Together, these studies provide new insights into the use of binding assays for molecular quantification and represent a rich resource for understanding the design of future assays
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 | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Wilson, Brandon Daniel |
|---|---|
| Degree supervisor | Bao, Zhenan |
| Degree supervisor | Soh, H. Tom |
| Thesis advisor | Bao, Zhenan |
| Thesis advisor | Soh, H. Tom |
| Thesis advisor | Davis, Ronald W. (Ronald Wayne), 1941- |
| Degree committee member | Davis, Ronald W. (Ronald Wayne), 1941- |
| Associated with | Stanford University, Department of Chemical Engineering. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Brandon D. Wilson |
|---|---|
| Note | Submitted to the Department of Chemical Engineering |
| Thesis | Thesis Ph.D. Stanford University 2020 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Brandon Daniel Wilson
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
Also listed in
Loading usage metrics...