A platform for constructing, evaluating, and screening bioconjugates on the yeast surface

被引:18
|
作者
Van Deventer, James A. [1 ,2 ,3 ]
Le, Doris N. [2 ,3 ]
Zhao, Jessie [2 ,3 ]
Kehoe, Haixing P. [1 ]
Kelly, Ryan L. [2 ,4 ]
机构
[1] Tufts Univ, Chem & Biol Engn Dept, 4 Colby St Room 148, Medford, MA 02155 USA
[2] MIT, Koch Inst Integrat Canc Res, 500 Main St,Bldg 76,Room 289, Cambridge, MA 02139 USA
[3] MIT, Dept Chem Engn, 500 Main St,Bldg 76,Room 289, Cambridge, MA 02139 USA
[4] MIT, Dept Biol Engn, 500 Main St,Bldg 76,Room 289, Cambridge, MA 02139 USA
来源
PROTEIN ENGINEERING DESIGN & SELECTION | 2016年 / 29卷 / 11期
基金
美国国家卫生研究院;
关键词
yeast display; noncanonical amino acids; click chemistry; antibodies; high throughput screening; AZIDE-ALKYNE CYCLOADDITION; EXPANDED GENETIC-CODE; UNNATURAL AMINO-ACIDS; DIRECTED EVOLUTION; IN-VIVO; RIBOSOMAL SYNTHESIS; SYNTHETIC BIOLOGY; BICYCLIC PEPTIDES; DISPLAY; STABILITY;
D O I
10.1093/protein/gzw029
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:485 / 493
页数:9
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