Peptides presented by cells
OwnerFrancis J. Carr, Graham Carter +1inventors
InventorsFrancis J. Carr, Graham Carter, Koen Hellendoorn
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Abstract
The present invention relates to methods to determine peptides presented on the surface of mammalian cells following addition to the cells of a protein. The present invention also relates to diagnostic tests based on the determination of such peptides or modified molecules resulting from determination of such peptides, such as pharmaceutical entities preferably having specific biological activity and reduced or enhanced immunogenicity when compared with the corresponding non-modified molecules. The methods according to this invention are preferably established with tools using mass spectroscopy (MS).
Claims34
A method for the development of a pharmaceutical protein or polypeptide or vaccine antigen having a specific biological activity and a reduced or enhanced immunogenicity than any non-modified protein or polypeptide having the same biological activity, by
- (i) contacting cells with said protein, or polypeptide, generating a repertoire of surface peptides on the cells or exosomal vehicles thereof, which is different from the repertoire of surface peptides displayed on reference cells which have not been contacted,
- (ii) analyzing said cells or exosomal vesicles thereof, for peptides bound on the surface of said cells or exosomal vesicles thereof and,
- (iii) assigning said peptides to the sequence of the pharmaceutical protein or polypeptide or vaccine antigen according to standard methods.
A method of claim I further comprising the following steps:
- (iv) modifying said peptides, in order to alter their binding to MHC molecules, and
- (v) constructing sequence variants of the final pharmaceutical protein or polypeptide by incorporating one or more of the modified peptide sequences within the sequence of the protein or polypeptide molecule according to standard methods.
A method according to any of the claims, wherein the pharmaceutical protein or polypeptide is modified such that one or more of said peptides are no longer bound after contacting cell with said protein or polypeptide.
A method for the development of a vaccine by
- (i) contacting cells with a protein or polypeptide or micro-organism having immunogenic activity, generating a repertoire of surface peptides on the said cells or exosomal vesicles thereof, which is different from the repertoire of surface peptides displayed on reference cells which have not been contacted,
- (ii) analyzing said cells or exosomal vesicles thereof, on the surface of which peptides are bound,
- (iii) assigning said peptides to the sequence of the protein or polypeptide according to standard methods, and
- (iv) constructing sequence variants of the final pharmaceutical vaccine by incorporating one or more of the peptides within the sequence of the vaccine molecule according to standard methods, wherein the analysis of said cells or exosomal vesicles therein is performed by using MS.
A method for the development of a diagnostic test by
- (i) analyzing appropriate human cells for surface peptides, and
- (ii) either, (a) producing a profile of peptides which appear on the cell surface and, optionally, comparing with other profiles to identify an abnormality or disease associated with the human cells, or, (b) determining the sequence of specific peptides on the cell surface as a means to determining specific peptides which might used to identify an abnormality or disease associated with the human cells.
A method for detecting peptides on the surface of cells or exosomal vesicles thereof which derive from a protein or polypeptide by
- (i) contacting cells with said protein, or polypeptide or a gene coding for this protein or polypeptide, generating a repertoire of surface peptides on that cells or exosomal vesicles thereof, which is different from the repertoire of surface peptides displayed on reference cells which have not been contacted,
- (ii) analyzing said cells or exosomal vesicles therein, on the surface of which said peptides are bound and
- (iii) assigning said peptides to the sequence of the protein or polypeptide according to standard methods, wherein the analysis if the cells or exosomal vesicles is performed by MS, preferably MALDI-MS.
Dependencies are inferred from the claim text.