Isogenica | Date: 2011-01-11
A method is provided for isolating protease resistant antimicrobial peptides (AMPs) from a peptide display library. A plurality of nucleic acid constructs that encode displayed peptides are expressed, resulting in the formation of a plurality of peptide-nucleic acid complexes, each complex comprising at least one displayed peptide associated with the corresponding nucleic acid construct encoding the displayed peptide. The complexes are exposed to at least one protease, to allow the proteolysis of protease-sensitive peptides, such that resistant peptides remain. The peptide-nucleic acid complexes are further exposed to a membrane composition to allow association of complexes that contain membrane-associating peptides. Complexes that remain unassociated with the membrane are removed; and membrane-associated complexes are recovered. The AMPs so characterised may be resistant to one or more protease enzymes and exhibit antimicrobial activity against one or more microbe.
Agency: Cordis | Branch: FP7 | Program: CP-FP | Phase: HEALTH.2011.1.1-1 | Award Amount: 5.55M | Year: 2012
We want to further develop our tools and technologies for high-throughput research, with the final goals being (I.) the particle-based combinatorial synthesis of 1 Mio different peptides on a glass slide for chemical costs of ~50 (KIT, CBL, MS, TUV), and (II.) the labelling-free parallel readout of binding affinities by a variant reflectometric interference spectroscopy method for ~10.000 peptide spots per cm(\2) when staining the array with an unlabeled protein (BIA, KIT). These tools provide the basis (III.) for a standardized, fast, and reliable high-throughput procedure that we want to develop in order to find high-affinity peptide binders against any pharmaceutically interesting target protein. Such a procedure might have an important impact in medicine and in the biotechnology industry. In order to achieve this goal, we will use display techniques that in combination with high-throughput sequencing typically will identify ~100.000 putative peptide binders per target protein (ISO). These will be synthesized in array format to validate binding to the target protein by an independent method (PPP, DKFZ). Next, based on binders from initial screens, many variant peptides are synthesized in high-density array format for iterative screens (PPP, DKFZ, KIT), whereby massive parallel labelling-free detection of binders pinpoints higher-affinity binders (BIA). In order to validate our novel high-throughput procedure, (IV.) we want to find high-affinity peptide binders against relevant target proteins (delivered by APO and OXF), and test these binders in biological assays (OXF, APO).
Isogenica | Date: 2010-10-21
The invention provides a method for making in vitro peptide expression libraries, and for the isolation of nucleotide sequences encoding peptides of interest, wherein the peptides or proteins are specifically associated with the DNA encoding them through non-covalent protein:DNA binding. The method describes ways of making the library itself, DNA molecules encoding the library and uses of the expression library.
Isogenica | Date: 2013-03-15
A method is disclosed for identifying a member of a peptide library that interacts with a target molecule in situ, the method including expressing immobilised nucleic acid molecules to produce the peptide library in a way that each member of the peptide library is immobilised on the nucleic acid molecule from which it was expressed; contacting the immobilised peptide library with the target molecule; and detecting an interaction between at least one member of the peptide library and the target molecule. The method further comprises sequencing the plurality of nucleic acid molecules in situ on the solid support, such that the at least one member of the peptide library that interacts with the target molecule can be immediately identified, at least by the sequence of the nucleic acid molecule from which it was expressed, without requiring additional or secondary analysis or characterising procedures in order to identify the useful members of the library. The target molecules may themselves be comprised within a second nucleic acid or peptide library.
Isogenica | Date: 2015-07-31
Diagnostic and therapeutic synthetic peptides, proteins, molecules, antibodies, polypeptides and polymers for the treatment of viral, metabolic, endocrine, musculoskeletal, cardiovascular, cardiopulmonary, genitourinary, sexual dysfunction, oncological, hepatological, ophthalmic, respiratory, neurological, gastrointestinal, hormonal, dermatological, psychiatric and immune system related diseases and disorders; biochemical and biological substances for medical diagnostics and analysis, namely, pharmaceutical preparations and substances for the diagnosis and analytics of viral, metabolic, endocrine, musculoskeletal, cardiovascular, cardiopulmonary, genitourinary, sexual dysfunction, oncological, hepatological, ophthalmic, respiratory, neurological, gastrointestinal, hormonal, dermatological, psychiatric and immune system related diseases and disorders. Scientific, pharmaceutical, medical, biological, biochemical, biotechnology and medical diagnostic research; biochemical and biotechnological engineering; biochemical engineering of peptides, proteins, molecules, antibodies, polypeptides and polymers. Licensing of intellectual property rights and of technology in the field of in-vitro screening and protein engineering; licensing of technology relating to discovery of peptides for therapeutic and non-therapeutic applications.