In this project, we propose to combine mRNA display (a protein design/evolution method) and high efficiency microfluidic sorting to create a new technology-microfluidic mRNA display-for the purpose of enabling design of peptides and proteins that can be used as protein capture reagents. We will develop and apply this powerful new technology toward creating a comprehensive reagent set aimed at the Hepatitis C virus (HCV) proteome. In this section, we begin by describing the existing state-of-the-art in 1) mRNA display-based peptide and protein design, 2) bead-based micromagnetic separations, and 3) give an introduction to the proteins expressed by the HCV that will be the targets of this work. This is followed by our description of how we will integrate these technologies to achieve our goal of high-throughput development of new protein capture reagents.
Title: Polypeptide Design with Proteomic Scope via Microfluidic mRNA Display Summary In this project, we propose to combine mRNA display (a protein design/evolution method) and high efficiency microfluidic sorting to create a new technology-microfluidic mRNA display-for the purpose of enabling design of peptides and proteins that can be used as protein capture reagents. We will develop and apply this powerful new technology toward creating a comprehensive reagent set aimed at the Hepatitis C virus (HCV) proteome. In this section, we begin by describing the existing state-of-the-art in 1) mRNA display-based peptide and protein design, 2) bead-based micromagnetic separations, and 3) give an introduction to the proteins expressed by the HCV that will be the targets of this work. This is followed by our description of how we will integrate these technologies to achieve our goal of high-throughput development of new protein capture reagents.
| Gross, Garrett G; Straub, Christoph; Perez-Sanchez, Jimena et al. (2016) An E3-ligase-based method for ablating inhibitory synapses. Nat Methods 13:673-8 |
| Jalali-Yazdi, Farzad; Lai, Lan Huong; Takahashi, Terry T et al. (2016) High-Throughput Measurement of Binding Kinetics by mRNA Display and Next-Generation Sequencing. Angew Chem Int Ed Engl 55:4007-10 |
| Jalali-Yazdi, Farzad; Takahashi, Terry T; Roberts, Richard W (2015) General, Label-Free Method for Determining K(d) and Ligand Concentration Simultaneously. Anal Chem 87:11755-62 |
| Wang, Jinpeng; Gong, Qiang; Maheshwari, Nupur et al. (2014) Particle display: a quantitative screening method for generating high-affinity aptamers. Angew Chem Int Ed Engl 53:4796-801 |
| Jalali-Yazdi, Farzad; Corbin, Jasmine M; Takahashi, Terry T et al. (2014) Robust, quantitative analysis of proteins using peptide immunoreagents, in vitro translation, and an ultrasensitive acoustic resonant sensor. Anal Chem 86:4715-22 |
| Olson, C Anders; Wu, Nicholas C; Sun, Ren (2014) A comprehensive biophysical description of pairwise epistasis throughout an entire protein domain. Curr Biol 24:2643-51 |
| Howell, Shannon M; Fiacco, Stephen V; Takahashi, Terry T et al. (2014) Serum stable natural peptides designed by mRNA display. Sci Rep 4:6008 |
| Gross, Garrett G; Junge, Jason A; Mora, Rudy J et al. (2013) Recombinant probes for visualizing endogenous synaptic proteins in living neurons. Neuron 78:971-85 |
| Mora, Rudy J; Roberts, Richard W; Arnold, Don B (2013) Recombinant probes reveal dynamic localization of CaMKII? within somata of cortical neurons. J Neurosci 33:14579-90 |
| Olson, C Anders; Nie, Jeff; Diep, Jonathan et al. (2012) Single-round, multiplexed antibody mimetic design through mRNA display. Angew Chem Int Ed Engl 51:12449-53 |
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