Date of Award

8-2020

Document Type

Campus Access Thesis

Degree Name

Master of Science (MS)

Department

Chemistry

First Advisor

Jason Evans

Second Advisor

Kimberly Hamad-Schifferli

Third Advisor

Marianna Torok

Abstract

A platform for evaluating the impact of a preformed protein corona on the sensitivity of a paper-based immunoassay utilizing gold nanoparticles was developed. Gold nanoparticles were synthesized and measured to be around 47 nm in diameter using Dynamic Light Scattering. The nanoparticles were coated with an anti-mouse IgG antibody and backfilled with a thiolated PEG to provide stability and prevent aggregation of the nanoparticles. These nanoparticles armed with IgG antibodies were used to perform a dipstick assay, designed to measure mouse IgG spiked into human serum at various concentrations. The Kdeff for this assay was measured to be 12.0 ± 2.4 pM. A proteomic analysis based on liquid chromatography-electrospray tandem mass spectrometry was performed on the protein corona formed when the nanoparticles were incubated in human serum. Human serum albumin was found to be an abundant protein present in the protein corona. The dipstick assay was performed using the immunoprobes that were first incubated in a solution of human serum albumin with the aim of preforming a protein corona of human serum albumin. Under these conditions, a Kdeff was measured to be 10.0 ± 3.5 pM. It was concluded that the preformation of a protein corona using human serum albumin had no significant impact on the performance of this dipstick assay.

Comments

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