(7e) Directed Evolution of Metalloproteinase Inhibitors Targeting ADAM-17
AIChE Annual Meeting
2021
2021 Annual Meeting
Food, Pharmaceutical & Bioengineering Division
Advances in Biosensors, Protein Production, and Protein Engineering
Sunday, November 7, 2021 - 4:42pm to 5:00pm
We used directed evolution and yeast surface display to screen a synthetic single chain antibody (anti-fluorescein scFv) library, previously engineered to reduce non-specific binding1, for improved binding to the ADAM-17 catalytic domain using fluorescent-activated cell sorting (FACS). DNA sequence analysis from each round of FACS, conducted via DNA sequence alignment bioinformatic tools, indicated that frequent antibody mutations within non-complementarity-determining regions (CDRs), light chain CDRs, and heavy chain CDR-2 are responsible for improving ADAM-17 binding. The individual scFv clones with improved binding affinity for ADAM-17 were further tested for MP binding affinities. This study set the stage for further engineering and design of protein scaffolds targeting MPs with high affinity and selectivity using directed evolution and rational design.
- Kelly, R. L., Le, D., Zhao, J., & Wittrup, K. D. (2018). Reduction of nonspecificity motifs in synthetic antibody libraries. Journal of Molecular Biology, 430(1), 119-130. doi:10.1016/j.jmb.2017.11.008