Coiled coil peptides as universal linkers for the attachment of recombinant proteins to polymer therapeutics
Language English Country United States Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
21863890
DOI
10.1021/bm200897b
Knihovny.cz E-resources
- MeSH
- Acrylamides chemistry MeSH
- Antigens, Neoplasm immunology metabolism MeSH
- Transformation, Bacterial MeSH
- Circular Dichroism MeSH
- Click Chemistry methods MeSH
- Dimerization MeSH
- Enzyme-Linked Immunosorbent Assay MeSH
- Escherichia coli MeSH
- Immunoconjugates chemistry immunology pharmacology MeSH
- Carbonic Anhydrase IX MeSH
- Carbonic Anhydrases immunology metabolism MeSH
- Carcinoma drug therapy enzymology immunology pathology MeSH
- Cloning, Molecular MeSH
- Humans MeSH
- Molecular Conformation MeSH
- Antibodies, Monoclonal chemistry genetics immunology MeSH
- Biomarkers, Tumor immunology metabolism MeSH
- Drug Carriers chemical synthesis pharmacology MeSH
- Oligopeptides chemical synthesis immunology pharmacology MeSH
- Plasmids MeSH
- Polyethylene Glycols chemistry MeSH
- Recombinant Proteins chemistry genetics immunology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Acrylamides MeSH
- Antigens, Neoplasm MeSH
- CA9 protein, human MeSH Browser
- Immunoconjugates MeSH
- Carbonic Anhydrase IX MeSH
- Carbonic Anhydrases MeSH
- M75 monoclonal antibody MeSH Browser
- Antibodies, Monoclonal MeSH
- N-(2-hydroxypropyl)methacrylamide MeSH Browser
- Biomarkers, Tumor MeSH
- Drug Carriers MeSH
- Oligopeptides MeSH
- Polyethylene Glycols MeSH
- Recombinant Proteins MeSH
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.
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