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Development of Protein Microarray Fabrication Methods for Analysis by Surface Plasmon Resonance Imaging

Creative Commons 'BY' version 4.0 license

This dissertation is the culmination of work toward the development of protein microarrays for surface plasmon resonance imaging (SPRI). Two main issues with protein microarrays for SPRI are addressed. First, we will address issues with introducing spatial selectivity into protein microarray fabrication strategies. We solve this problem first by physical separation of protein synthesis in a microwell array, followed by contact printing onto a functionalized SPRI chip. In our second solution for this spatial selectivity problem, we introduce a novel protein adsorption chemistry based on zinc finger/DNA affinity. This chemistry is explored as a potential adsorption chemistry for self-assembled protein microarrays.

The second issue we will address is lack of unnatural amino acids in current protein microarray fabrication methods. Unnatural amino acids allow users to introduce precise chemical control into proteins, but despite their utility, they have not been introduced into current fabrication strategies because of their biosynthetic difficulty. We address this issue by introducing an on chip amber suppression strategy, using a ribozyme based tRNA misacylation system.

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