19 research outputs found
Re-assessment of YAP1 and MCR1 contributions to inhibitor tolerance in robust engineered Saccharomyces cerevisiae fermenting undetoxified lignocellulosic hydrolysate
Diseño y validaciĂłn de un instrumento para evaluar el conocimiento tĂĄcito de lĂderes militares
Combining Phage and Yeast Cell Surface Antibody Display to Identify Novel Cell Type-Selective Internalizing Human Monoclonal Antibodies
Leisure, Free Time and Well-Being of 10Â Years Old Children Living in Buenos Aires Province, Argentina
Ribozyme-Based Aminoglycoside Switches of Gene Expression Engineered by Genetic Selection in S. cerevisiae
Functional Heterologous Protein Expression by Genetically Engineered Probiotic Yeast Saccharomyces boulardii
Massively parallel de novo protein design for targeted therapeutics
De novo protein design holds promise for creating small stable proteins with shapes customized to bind therapeutic targets. We describe a massively parallel approach for designing, manufacturing and screening mini-protein binders, integrating large-scale computational design, oligonucleotide synthesis, yeast display screening and next-generation sequencing. We designed and tested 22,660 mini-proteins of 37-43 residues that target influenza haemagglutinin and botulinum neurotoxin B, along with 6,286 control sequences to probe contributions to folding and binding, and identified 2,618 high-affinity binders. Comparison of the binding and non-binding design sets, which are two orders of magnitude larger than any previously investigated, enabled the evaluation and improvement of the computational model. Biophysical characterization of a subset of the binder designs showed that they are extremely stable and, unlike antibodies, do not lose activity after exposure to high temperatures. The designs elicit little or no immune response and provide potent prophylactic and therapeutic protection against influenza, even after extensive repeated dosing