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Engineering High Affinity Protein–Protein Interactions Using a High-Throughput Microcapillary Array Platform
- Source :
- ACS Chemical Biology; February 2017, Vol. 12 Issue: 2 p336-341, 6p
- Publication Year :
- 2017
-
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.
Details
- Language :
- English
- ISSN :
- 15548929 and 15548937
- Volume :
- 12
- Issue :
- 2
- Database :
- Supplemental Index
- Journal :
- ACS Chemical Biology
- Publication Type :
- Periodical
- Accession number :
- ejs41353410
- Full Text :
- https://doi.org/10.1021/acschembio.6b00794