Protein and peptide microarrays have exploded onto the biotechnology scene over the past decade. These microarrays, coupled with surface plasmon resonance imaging (SPRI), are potentially powerful tools for the simultaneous, high-throughput analysis of an enormous number of samples per experiment, which grant scientists an unprecedented combination of speed and volume in proteomic data collection. Unnatural proteins however, are rarely incorporated into existing fabrication strategies despite their massive potential in therapeutics and proteomics among other applications. Our strategy for the inclusion of unnatural proteins into microarrays suitable for SPRI will be described in this presentation.<br/><br/>We use an on-chip fabrication strategy for the creation of these unnatural protein microarrays. Parallel <i>in vitro</i> protein synthesis is performed in a microwell array. In each microwell, a single residue variation is introduced at the translation level via amber codon suppression. A misacylated tRNA is used to introduce an unnatural amino acid at a specific location in the protein. The resulting protein microwell array then serves as a stamp for His-tag immobilization of proteins onto an SPRI chip via contact printing. This on-chip synthesis method has the potential for quickly and efficiently creating complex unnatural protein microarrays suitable for SPRI analysis.

Templated biosynthesis of unnatural and natural proteins for surface plasmon resonance imaging (SPRI)

251st National Meeting (2016)

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.