AI Article Synopsis
- Affinity maturation involves repeated cycles of creating protein libraries and screening them to find variants with better binding to specific targets.
- The μSCALE platform allows for the rapid screening of millions of protein variants based on their binding ability or activity.
- In a recent study, μSCALE was used successfully to enhance the binding of the Axl receptor to its ligand Gas6, achieving a significant improvement in binding strength within just two weeks.
Article 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.
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| http://dx.doi.org/10.1021/acschembio.6b00794 | DOI Listing |
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