Turechek, Luke J.2019-12-102018-112018-07-162018Turechek, Luke J. 2018. Adnectin Ligand Affinity Optimization With Dual Combinatorial Positional Scan Libraries. Master's thesis, Harvard Extension School.http://nrs.harvard.edu/urn-3:HUL.InstRepos:42004030Discovery of antibody and alternative-scaffold ligands such as Adnectins to a protein target of interest has long been important for assay development and drug discovery. Affinity maturation of candidates with poor binding kinetics can hinder development in a fast-paced environment, with traditional methods being very time consuming, and faster methods involving single point mutation scanning being unlikely to yield highly improved variants. This thesis describes a method for the rapid discovery of affinity-matured variants of an EGFR-binding Adnectin using a chip oligo-based dual mutant library design in combination with mRNA display and next-generation sequencing.application/pdfadnectinligand discoverymRNA displaynext-generation sequencingNGSaffinity maturationEGFRThesis or Dissertation2019-12-10