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 confirmed by CD spectroscopy. 5-Azidopentanoic acid was connected to the N-terminus of peptide E via a short poly(ethylene glycol) spacer. The 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 thereby mediates a noncovalent linkage between the polymer drug carrier and the protein. The recombinant scFv antibody fragment was selected as a targeting ligand against carbonic anhydrase IX-a marker overexpressed by tumor cells of various human carcinomas. The antigen binding affinity of the polymer-scFv complex was confirmed by ELISA. This approach offers a well-defined, specific, and nondestructive universal method for the preparation of protein (antibody)-targeted polymer drug and gene carriers designed for cell-specific delivery.

journal_name

Biomacromolecules

journal_title

Biomacromolecules

authors

Pechar M,Pola R,Laga R,Ulbrich K,Bednárová L,Maloň P,Sieglová I,Král V,Fábry M,Vaněk O

doi

10.1021/bm200897b

subject

Has Abstract

pub_date

2011-10-10 00:00:00

pages

3645-55

issue

10

eissn

1525-7797

issn

1526-4602

journal_volume

12

pub_type

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