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Coiled coil peptides as universal linkers for the attachment of recombinant proteins to polymer therapeutics
- Source :
- Biomacromolecules. 12(10)
- Publication Year :
- 2011
-
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.
- Subjects :
- Immunoconjugates
Polymers and Plastics
Stereochemistry
Molecular Conformation
Bioengineering
Peptide
Enzyme-Linked Immunosorbent Assay
law.invention
Polyethylene Glycols
Biomaterials
law
Antigens, Neoplasm
Materials Chemistry
Biomarkers, Tumor
Escherichia coli
Humans
Cloning, Molecular
Carbonic Anhydrase IX
Carbonic Anhydrases
Coiled coil
chemistry.chemical_classification
Oligopeptide
Acrylamides
Drug Carriers
Chemistry
Circular Dichroism
Carcinoma
Antibodies, Monoclonal
Ligand (biochemistry)
Random coil
Recombinant Proteins
Biochemistry
Click chemistry
Recombinant DNA
Click Chemistry
Transformation, Bacterial
Drug carrier
Dimerization
Oligopeptides
Plasmids
Subjects
Details
- ISSN :
- 15264602
- Volume :
- 12
- Issue :
- 10
- Database :
- OpenAIRE
- Journal :
- Biomacromolecules
- Accession number :
- edsair.doi.dedup.....060b01b633f7999d83b06a1cc0d55fa0