title: A platform for constructing, evaluating, and screening bioconjugates on the yeast surface.
abstract: The combination of protein display technologies and noncanonical amino acids (ncAAs) offers unprecedented opportunities for the high throughput discovery and characterization of molecules suitable for addressing fundamental and applied problems in biological systems. Here we demonstrate that ncAA-compatible yeast display facilitates evaluations of conjugation chemistry and stability that would be challenging or impossible to perform with existing mRNA, phage, or E. coli platforms. Our approach enables site-specific introduction of ncAAs into displayed proteins, robust modification at azide-containing residues, and quantitative evaluation of conjugates directly on the yeast surface. Moreover, screening allows for the selective enrichment of chemically modified constructs while maintaining a genotype-phenotype linkage with encoded azide functionalities. Thus, this platform is suitable for the high throughput characterization and screening of libraries of chemically modified polypeptides for therapeutic lead discovery and other biological applications.
authors: Chemical and Biological Engineering Department, Tufts University, 4 Colby Street Room 148, Medford, MA 02155, United States of America.|Koch Institute for Integrative Cancer Research.|Department of Chemical Engineering.
authors: James A
authors: JA
authors: Van Deventer
authors: Koch Institute for Integrative Cancer Research.|Department of Chemical Engineering.
authors: Doris N
authors: DN
authors: Le
authors: Koch Institute for Integrative Cancer Research.|Department of Chemical Engineering.
authors: Jessie
authors: J
authors: Zhao
authors: Chemical and Biological Engineering Department, Tufts University, 4 Colby Street Room 148, Medford, MA 02155, United States of America.
authors: Haixing P
authors: HP
authors: Kehoe
authors: Koch Institute for Integrative Cancer Research.|Department of Biological Engineering, Massachusetts Institute of Technology, 500 Main Street, Building 76 Room 289, Cambridge, MA 02139, United States of America.
authors: Ryan L
authors: RL
authors: Kelly
keywords: antibodies; click chemistry; high throughput screening; noncanonical amino acids; yeast display