Effective screening for hypolipidemic bile acid - binding peptides by hierarchical clustering - based catalog array

Abstract

Peptide arrays, which are high-throughput combinatorial screening platforms, have a limited capacity for exploring vast amino acid sequence combinations. To mitigate such limitations, we propose the concept of "catalog array," comprised of bioinformatically enriched minimal peptides representing maximal physicochemical variety. For this method, hierarchical clustering was introduced to group and select peptides with similar physiochemical properties to be synthesized on the array. Indeed, clustering determined that all 3-mer peptides could be narrowed to 47 groups using the 13 amino acid indices. We subsequently evaluated the validity of our array by screening for bile acid-binding proteins. As a target bio-functional peptide, bile acid-binding peptides have recently gained attention due to their hypolipidemic properties. Notably, the catalog array provided up to four-fold higher screening efficacy (23.2% efficacy) over simple random screening (6.9%), enabling the identification of peptides with greater bile acid-binding capacities than the previously reported 6-mer peptide VAWWMY.