Date of Award

Spring 2024

Document Type

Thesis

Degree Name

Bachelor of Arts (BA)

Department

Chemistry

First Advisor

Maryuri Roca

Second Advisor

Madushi Raththagala

Abstract

Western blot is an important protein analysis assay enabling protein detection by visualizing the antibody specific antigen-antibody interaction. As a commonly used assay, western blot is taught in upper-level biochemistry and molecular biology students. However, due to its exhaustive process and high cost of antibodies, a modification to western blot is proposed using gold or silver nanoparticles as a tag to the primary antibody for lysozyme. In this experiment, invisible anti-lysozyme antibodies were conjugated to colloidal gold and silver nanoparticles via photochemical immobilization technique, effectively facilitating a physically observable band on the western blot. To eliminate false positives by interactions between immobilized proteins and the nanoparticles, exposed surfaces of metal were covered with different passivating molecules, of those denatured protein show the most promising results. Gold nanoparticles were modeled for silver nanoparticle tags due to its superior stability in buffers compared to its silver counterpart. However, silver nanoparticles are more convenient due to their ease of synthesis and cost advantages. Both gold and silver labeled antibody enables cost and time optimization to western blot as it does not require a secondary antibody nor developer like horse radish peroxidase.

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