Directed Evolution of Glycopeptides Using mRNA Display.

摘要: Abstract Directed evolution is a useful method for the discovery of nucleic acids, peptides, or proteins that have desired binding abilities functions. Because abundance and importance glycosylation in nature, directed glycopeptides glycoproteins also highly desirable. However, common platforms such as phage-, yeast-, mammalian-cell display are limited these applications by several factors. Glycan structure at each site not genetically encoded, yeast mammalian cells produce heterogeneous mixture glycoforms on protein. Although yeast, Escherichia coli can be engineered to homogenous glycoform all sites, there just few specific glycan structures readily accessed this manner. Recently, we reported novel system glycopeptide libraries, which could principle decorated with any glycan. Our combines vitro peptide selection mRNA unnatural amino acid incorporation chemical attachment synthetic oligosaccharides. Here, provide an updated optimized protocol method, designed create libraries containing ~ 10 13 sequences select them target binding. The described here HIV broadly neutralizing monoclonal antibody 2G12; 2G12 binds cluster high-mannose oligosaccharides envelope glycoprotein gp120; mimic epitope may vaccine applications. This expected applicable other types glycans targets interest glycobiology.