Coiled coil peptides as universal linkers for the attachment of recombinant proteins to polymer therapeutics
Abstract
We have designed, synthesized, and characterized peptides containing four repeats of the sequences VAALEKE (peptide E) or VAALKEK (peptide K). While the peptides alone adopt in aqueous solutions a random coil conformation, their equimolar mixture forms heterodimeric coiled coils as conWe prepared peptides containing four repeats of the sequences VAALEKE (peptide E) or VAALKEK (peptide K).
While the peptides alone adopt in aqueous solutions a random coil conformation, their equimolar mixture forms heterodimeric coiled coils as confirmed by CD spectroscopy. A terminal azide group enabled conjugation of the peptide with a synthetic drug carrier based on the N-(2-hydroxypropyl)methacrylamide copolymer containing propargyl groups using "click" chemistry.
When incorporated into the polymer drug carrier, peptide E formed a stable noncovalent complex with peptide K belonging to a recombinant single-chain fragment (scFv) of the M75 antibody. The complex mediated a noncovalent linkage between the polymer drug carrier and the protein.
The antigen binding affinity of the polymerMINUS SIGN scFv complex was confirmed by ELISA. This approach offers a well-defined, specific, and nondestructive universal method for the preparation of protein-targeted polymer drug carriers.