Lasso-grafting of macrocyclic peptide pharmacophores yields multi-functional proteins

Abstract Protein engineering has great potential for devising multifunctional recombinant proteins to serve as next-generation protein therapeutics, but it often requires drastic modifications of the parental scaffolds e.g., additional domains at N/C-terminus or replacement a domain by another. A discovery platform system, called RaPID (Random non-standard Peptides Integrated Discovery) enabled rapid small de novo macrocyclic peptides that bind target with high binding specificity and affinity. Capitalizing on optimized properties RaPID-derived peptides, here we show pharmacophore sequences can be readily implanted into surface-exposed loops maintain both peptide function(s) host function. We refer this method lasso-grafting demonstrate endow specific capacity toward various receptors diverse set includes IgG, serum albumin, even capsid adeno-associated virus, enabling us rapidly formulate produce bi-, tri-, tetra-specific binder molecules.