Date of Award

2019

Degree Name

Chemistry

College

College of Science

Type of Degree

M.S.

Document Type

Thesis

First Advisor

Dr. Michael Norton, Committee Chairperson

Second Advisor

Dr. Scott Day

Third Advisor

Dr. Bin Wang

Abstract

Lateral flow assays (LFA) are used for point-of-care qualitative diagnostics of an analyte of interest, often in non-laboratory environments. Traditionally, the format of a lateral flow assay is to utilize immobilized antibodies on a membrane as the capture probe in conjunction with a reporting immunological recognition system for an analyte captured between them in a sandwich format. However, there are several shortcomings of antibodies which recommend their replacement with other recognition elements, if possible. The research described in this thesis was directed toward using several of the inherent properties of DNA based Origami nanostructures to enable the construction of DNA based LFAs. DNA Origami constructs consist of a scaffold strand, staple strands (to direct for the folding of helices to conform to the specification of the design) and modified staple strands. Programmable functionalities include specific DNA structures, nanoparticles, proteins and a wide range of chemical modifications. The three types of modifications which were investigated as potential analyte binding mechanisms included: 1) ssDNA hybridization, 2) Tris-Nitrilotriacetic Acid (Tris-NTA)-his tagged protein binding and 3) binding to Thrombin through aptamers. The capture probe of the lateral flow assay was immobilized on an analytical Nitrocellulose membrane. The mobile reporting probe DNA Origami construct was modified to enable binding of Gold nanoparticles which provide a mechanism for development of a visible, characteristic red report line at the test site, indicative of a positive test result.

Subject(s)

DNA -- Structure.

Colloidal gold -- Research.

Nanostructured materials -- Research.

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