Engineering High Affinity Protein-Protein Interactions Using a High-Throughput Microcapillary Array Platform.
ACS Chem Biol
; 12(2): 336-341, 2017 02 17.
Article
in En
| MEDLINE
| ID: mdl-27997117
ABSTRACT
Affinity maturation of protein-protein interactions requires iterative rounds of protein library generation and high-throughput screening to identify variants that bind with increased affinity to a target of interest. We recently developed a multipurpose protein engineering platform, termed µSCALE (Microcapillary Single Cell Analysis and Laser Extraction). This technology enables high-throughput screening of libraries of millions of cell-expressing protein variants based on their binding properties or functional activity. Here, we demonstrate the first use of the µSCALE platform for affinity maturation of a protein-protein binding interaction. In this proof-of-concept study, we engineered an extracellular domain of the Axl receptor tyrosine kinase to bind tighter to its ligand Gas6. Within 2 weeks, two iterative rounds of library generation and screening resulted in engineered Axl variants with a 50-fold decrease in kinetic dissociation rate, highlighting the use of µSCALE as a new tool for directed evolution.
Full text:
1
Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protein Engineering
/
Proteins
Language:
En
Journal:
ACS Chem Biol
Year:
2017
Document type:
Article
Affiliation country:
Estados Unidos