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Publication Number
US-7888316-B2
Publication Date
2011-02-15
Expiration Date
Abstract
Disclosed herein are polypeptide multilayer films comprising a hybrid polypeptide comprising a first polypeptide segment and a second segment, the two segments being covalently joined by one or more non-peptidic linkages. The first segment comprises a polypeptide having a magnitude of net charge per residue of greater than or equal to 0.4, and a length of greater than or equal to about 12 amino acid residues. The second segment comprises a polypeptide or another polyelectrolyte.
Core Innovation
The invention relates to multilayer film formed from two or more layers of polyelectrolytes with adjacent layers comprising oppositely charged polyelectrolytes, where one layer includes a hybrid polypeptide. The hybrid polypeptide comprises a first polypeptide segment covalently joined to a second polypeptide segment by one or more non-peptidic linkages, and the covalent junction supports multilayer assembly with oppositely charged polyelectrolytes.
The first polypeptide segment provides a polypeptide with a magnitude of net charge per residue of at least a specified threshold and a length of at least a specified number of amino acid residues, which supports adsorption and solubility in pH 4–10. The second polypeptide segment provides a functional region comprising specified lengths and selected from a set of functional sequences and domains, distinguishing a surface adsorption region from functional regions and domains.
The functional region is enumerated to include cysteine-containing sequences, extracellular matrix recognition sequences, phosphotyrosine binding domains, Src homology 2 domains, N-linked glycosylation substrate sequences, O-linked glycosylation substrate sequences, protease recognition sites, antimicrobial peptide sequences, BAG domains, and T-cell epitopes, including combinations thereof. The disclosure further includes film stabilization via disulfides, modular synthesis and non-peptidic linker chemistries, and examples in which the hybrid polypeptide supports epitope presentation, receptor or ligand interactions, and redox-responsive behavior using non-peptidic linkers and second segments.
Claims Coverage
The independent claim is clm-00001. The inventive features involve 6 inventive features: an oppositely charged polyelectrolyte multilayer film, a hybrid polypeptide in a first layer with non-peptidic covalent junction chemistry, specific charge and length constraints on a first segment, a bounded-size functional region selection for a second segment, non-peptidic end-to-end segment connectivity, and specified charge and molecular-weight requirements for the opposing second-layer polyelectrolyte.
Oppositely charged polyelectrolyte multilayer film
A multilayer film comprising two or more layers of polyelectrolytes wherein adjacent layers comprise oppositely charged polyelectrolytes.
Hybrid polypeptide layer with non-peptidic covalent junction
The first layer polyelectrolyte comprises a hybrid polypeptide comprising a first polypeptide segment and a second polypeptide segment covalently joined by one or more non-peptidic linkages.
Charged adsorption segment defined by net charge per residue and length
The first polypeptide segment comprises a polypeptide having a magnitude of net charge per residue of greater than or equal to 0.4 and a length of greater than or equal to about 12 amino acid residues.
Functional region selection for second segment
The second polypeptide segment comprises a functional region comprising 3 to about 250 amino acids selected from the group consisting of a cysteine-containing sequence, a sequence for extracellular matrix recognition, a phosphotyrosine binding domain, a Src homology 2 domain, an N-linked glycosylation substrate sequence, an O-linked glycosylation substrate sequence, a protease recognition site, an antimicrobial peptide sequence, a BAG domain, a T-cell epitope, or a combination thereof.
Non-peptidic end-to-end segment connectivity
The N-terminus of the first polypeptide segment is joined to the C-terminus of the second polypeptide segment, the N-terminus of the first polypeptide segment is joined to the N-terminus of the second polypeptide segment, the C-terminus of the first polypeptide segment is joined to the C-terminus of the second polypeptide segment, or the C-terminus of the first polypeptide segment is joined to the N-terminus of the second polypeptide segment by the one or more non-peptidic linkages.
Opposing second layer polyelectrolyte with defined molecular weight and charge
The second layer polyelectrolyte comprises a polycationic material or a polyanionic material having a molecular weight of greater than 1,000, at least 5 charges per molecule, and a charge opposite that of the first layer polypeptide.
Across the independent claim, the coverage is directed to multilayer polyelectrolyte films with adjacent oppositely charged layers, where the first layer includes a hybrid polypeptide formed by covalently joining a charged and length-defined first segment to a functional-region second segment via one or more non-peptidic linkages, and where the opposing second layer is a polycationic or polyanionic material meeting defined molecular-weight and charge characteristics.
Stated Advantages
Not explicitly described in patent.
Documented Applications
Not explicitly described in patent.
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