Abstract: The present invention relates to polypeptides that are resistant to degradation in the gastrointestinal tract and that bind to a target (i.e. a target ligand). In particular, it provides a mutant Kunitz-type soybean trypsin inhibitor (SBTI) family polypeptide comprising two or more amino acid mutations compared to the corresponding unmutated (e.g. wild-type) SBTI family polypeptide, wherein the mutant SBTI family polypeptide comprises: (i) one or more amino acid mutations in a first domain corresponding to positions 22-25 of SEQ ID NO: 1; and (ii) one or more amino acid mutations in a second domain corresponding to positions 47-50 of SEQ ID NO: 1, wherein the mutant SBTI family polypeptide: (a) binds selectively to a ligand that does not bind to the corresponding unmutated (e.g. wild-type) SBTI family polypeptide; and (b) is resistant to cleavage by pepsin.

Abstract: The present invention relates to a polypeptide that forms one part of a two-part linker in which the polypeptide spontaneously forms an isopeptide bond with a peptide tag, the second part of the two-part linker. Nucleic acid molecules encoding the polypeptide, vectors comprising said nucleic acid molecules, and host cells comprising said vectors and nucleic acid molecules are also provided. A kit comprising said two-part linker (i.e. peptide tag and polypeptide binding partner), and/or nucleic acid molecules/vectors is also provided. A method of producing the polypeptide and the uses of the polypeptide of the invention are also provided.

Abstract: The present invention relates to a two-part linker comprising a peptide tag (peptide) and a polypeptide (protein) that is capable of spontaneously forming an isopeptide bond, particularly wherein: a) said peptide comprises an amino acid sequence as set forth in SEQ ID NO: 1, wherein: (i) X at position 1 is arginine or no amino acid; (ii) X at position 2 is glycine or no amino acid; (iii) X at position 5 is histidine or threonine; (iv) X at position 11 is alanine, glycine or valine; and (v) X at position 14 is arginine or lysine, wherein when X at position 1 is no amino acid, X at position 2 is no amino acid; and b) said polypeptide comprises: i) an amino acid sequence as set forth in SEQ ID NO: 2; ii) a portion of (i) comprising an amino acid sequence as set forth in SEQ ID NO: 101; iii) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 2, wherein said amino acid sequence comprises a lysine at position 34, a glutamic acid at position 80 and one or more of the

Abstract: The present invention relates to a system for generating intermolecular covalent bonds (e.g. amide, e.g. isopeptide bonds) between polypeptides. In particular, it provides the use of a chimeric protein to generate an anhydride group on a polypeptide for the formation of a covalent bond, wherein the chimeric protein comprises (i) a domain comprising the polypeptide and (ii) a domain comprising a self-processing module that contains an N-terminal dipeptide of aspartate or glutamate and proline (D/E-P), wherein (i) and (ii) are linked by a peptide bond between the aspartate or glutamate residue at the N-terminus of (ii) and the amino acid at the C-terminus of (i) and wherein the self-processing module cleaves the peptide bond between the proline residue and the aspartate or glutamate residue in the self-processing module to release the polypeptide and generate the anhydride group on the aspartate or glutamate residue.

Abstract: This invention relates to preparations comprising adenoviral vectors with modified capsid proteins. These modified capsid proteins enable customisable decoration of the adenoviral vector to be performed, enabling diverse applications from personalised cancer vaccines to targeted gene therapy vectors, and mixtures of the same. In particular, the adenoviral vectors with modified capsid proteins may be modified in the hexon and/or pIX capsid proteins. The invention makes use of peptide pairs to provide a “primed” adenovirus which is ready for decoration.

Abstract: The present invention relates to a polypeptide that forms one part of a two-part linker in which the polypeptide spontaneously forms an isopeptide bond with a peptide tag, the second part of the two-part linker. Nucleic acid molecules encoding the polypeptide, vectors comprising said nucleic acid molecules, and host cells comprising said vectors and nucleic acid molecules are also provided. A kit comprising said two-part linker (i.e. peptide tag and polypeptide binding partner), and/or nucleic acid molecules/vectors is also provided. A method of producing the polypeptide (binding partner) and the uses of the polypeptide of the invention are also provided.

Abstract: The present invention relates to a two-part linker comprising a peptide tag (peptide) and a polypeptide (protein) that is capable of spontaneously forming an isopeptide bond, particularly wherein: a) said peptide comprises an amino acid sequence as set forth in SEQ ID NO: 1, wherein: (i) X at position 1 is arginine or no amino acid; (ii) X at position 2 is glycine or no amino acid; (iii) X at position 5 is histidine or threonine; (iv) X at position 11 is alanine, glycine or valine; and (v) X at position 14 is arginine or lysine, wherein when X at position 1 is no amino acid, X at position 2 is no amino acid; and b) said polypeptide comprises: i) an amino acid sequence as set forth in SEQ ID NO: 2; ii) a portion of (i) comprising an amino acid sequence as set forth in SEQ ID NO: 101; iii) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 2, wherein said amino acid sequence comprises a lysine at position 34, a glutamic acid at position 80 and one or more of the

Abstract: The present invention relates to an affinity purification system that utilises a polypeptide comprising: (i) an amino acid sequence as set forth in SEQ ID NO: 1, wherein X at position 79 is selected from alanine, glycine, serine, asparagine, or threonine; (ii) a portion of (i) comprising an amino acid sequence as set forth in SEQ ID NO: 2, wherein X at position 56 is selected from alanine, glycine, serine, asparagine or threonine; (iii) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 1, wherein the polypeptide comprises alanine, glycine, serine, asparagine or threonine at a position equivalent to position 79 of SEQ ID NO: 1; or (iv) a portion of (iii) comprising an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 2, wherein the polypeptide comprises alanine, glycine, serine, asparagine or threonine at a position equivalent to position 56 of SEQ ID NO: 2, wherein the polypeptide binds selectively and reversibly

Abstract: The present invention relates to a two-part linker comprising a peptide tag (peptide) and a polypeptide (protein) that is capable of spontaneously forming an isopeptide bond, particularly wherein: a) said peptide comprises an amino acid sequence as set forth in SEQ ID NO: 1, wherein: (i) X at position 1 is arginine or no amino acid; (ii) X at position 2 is glycine or no amino acid; (iii) X at position 5 is histidine or threonine; (iv) X at position 11 is alanine, glycine or valine; and (v) X at position 14 is arginine or lysine, wherein when X at position 1 is no amino acid, X at position 2 is no amino acid; and b) said polypeptide comprises: i) an amino acid sequence as set forth in SEQ ID NO: 2; ii) a portion of (i) comprising an amino acid sequence as set forth in SEQ ID NO: 101; iii) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 2, wherein said amino acid sequence comprises a lysine at position 34, a glutamic acid at position 80 and one or more of the

Abstract: The present invention relates to a polypeptide that is capable of promoting the covalent conjugation of two peptide tags or linkers and in particular to a polypeptide comprising: a) an amino acid sequence as set forth in SEQ ID NO: 1; or b) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 1, wherein said amino acid sequence comprises a glutamic acid at position 61 and one or more of the following: 1) proline at position 66; 2) proline at position 95; 3) glycine at position 96; and 4) valine at position 97, wherein the specified amino acid residues are at positions equivalent to the positions in SEQ ID NO: 1 and wherein said polypeptide is capable of promoting the formation of an isopeptide bond between the lysine residue at position 9 of SEQ ID NO: 2 and the asparagine residue at position 17 of SEQ ID NO: 3.

Abstract: The present invention relates to a two-part linker comprising a peptide tag (peptide) and a polypeptide (protein) that is capable of spontaneously forming an isopeptide bond, particularly wherein: a) said peptide comprises an amino acid sequence as set forth in SEQ ID NO: 1, wherein: (i) X at position 1 is arginine or no amino acid; (ii) X at position 2 is glycine or no amino acid; (iii) X at position 5 is histidine or threonine; (iv) X at position 11 is alanine, glycine or valine; and (v) X at position 14 is arginine or lysine, wherein when X at position 1 is no amino acid, X at position 2 is no amino acid; and b) said polypeptide comprises: i) an amino acid sequence as set forth in SEQ ID NO: 2; ii) a portion of (i) comprising an amino acid sequence as set forth in SEQ ID NO: 101; iii) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 2, wherein said amino acid sequence comprises a lysine at position 34, a glutamic acid at position 80 and one or more of the

Abstract: The present invention relates to a polypeptide that is capable of promoting the covalent conjugation of two peptide tags or linkers and in particular to a polypeptide comprising: a) an amino acid sequence as set forth in SEQ ID NO: 1; or b) an amino acid sequence with at least 80% sequence identity to a sequence as set forth in SEQ ID NO: 1, wherein said amino acid sequence comprises a glutamic acid at position 61 and one or more of the following: 1) proline at position 66; 2) proline at position 95; 3) glycine at position 96; and 4) valine at position 97, wherein the specified amino acid residues are at positions equivalent to the positions in SEQ ID NO: 1 and wherein said polypeptide is capable of promoting the formation of an isopeptide bond between the lysine residue at position 9 of SEQ ID NO: 2 and the asparagine residue at position 17 of SEQ ID NO: 3.

Abstract: Described herein is a protein capable of spontaneously forming an isopeptide bond for the development of a peptide tag and binding partner pair wherein the peptide tag and binding partner are capable of covalently binding to each other via an isopeptide bond. Also provided is a method for developing a peptide tag and binding partner pair which are capable of covalently binding to each other based on a protein which is capable of spontaneously forming an isopeptide bond. Additionally provided are peptide tag and binding partner pairs which are obtainable from isopeptide proteins. Further, specifically developed peptide tags and binding partners are described, together with nucleic acid molecules and vectors which encode those peptides or proteins.

Abstract: A method of producing a fusion protein is provided. The method includes: a) contacting a first protein with a second protein under conditions that enable the formation of an isopeptide bond between said proteins to form a linked protein; and b) contacting the linked protein from (a) with a third protein under conditions that enable the formation of an isopeptide bond between said third protein and said linked protein to form a fusion protein. Peptide linkers and the use of orthogonal pairs of said linkers in the synthesis of fusion proteins are also provided. Recombinant proteins comprising said linkers, nucleic acid molecules encoding said proteins and linkers, vectors comprising said nucleic acid molecules and host cells comprising said vectors and nucleic acid molecules are also contemplated.

Abstract: Described herein is a protein capable of spontaneously forming an isopeptide bond for the development of a peptide tag and binding partner pair wherein the peptide tag and binding partner are capable of covalently binding to each other via an isopeptide bond. Also provided is a method for developing a peptide tag and binding partner pair which are capable of covalently binding to each other based on a protein which is capable of spontaneously forming an isopeptide bond. Additionally provided are peptide tag and binding partner pairs which are obtainable from isopeptide proteins. Further, specifically developed peptide tags and binding partners are described, together with nucleic acid molecules and vectors which encode those peptides or proteins.

Abstract: Described herein is-a protein capable of spontaneously forming an isopeptide bond for the development of a peptide tag and binding partner pair wherein the peptide tag and binding partner are capable of covalently binding to each other via an isopeptide bond. Also provided is a method for developing a peptide tag and binding partner pair which are capable of covalently binding to each other based on a protein which is capable of spontaneously forming an isopeptide bond. Additionally provided are peptide tag and binding partner pairs which are obtainable from isopeptide proteins. Further, specifically developed peptide tags and binding partners are described, together with nucleic acid molecules and vectors which encode those peptides or proteins.

Abstract: The present invention provides a method of producing a fusion protein, said method comprising: a) contacting a first protein with a second protein under conditions that enable the formation of an isopeptide bond between said proteins, wherein said first protein and said second protein each comprise a peptide linker, wherein said peptide linkers are a pair of peptide linkers which react to form an isopeptide bond that links said first protein to said second protein to form a linked protein; and b) contacting the linked protein from (a) with a third protein under conditions that enable the formation of an isopeptide bond between said third protein and said linked protein, wherein said third protein comprises a peptide linker which reacts with a further peptide linker in the linked protein from (a), and wherein said peptide linkers are a pair of peptide linkers that react to form an isopeptide bond that links said third protein to said linked protein to form a fusion protein, wherein said pair of peptide linkers

Abstract: Described herein is-a protein capable of spontaneously forming an isopeptide bond for the development of a peptide tag and binding partner pair wherein the peptide tag and binding partner are capable of covalently binding to each other via an isopeptide bond. Also provided is a method for developing a peptide tag and binding partner pair which are capable of covalently binding to each other based on a protein which is capable of spontaneously forming an isopeptide bond. Additionally provided are peptide tag and binding partner pairs which are obtainable from isopeptide proteins. Further, specifically developed peptide tags and binding partners are described, together with nucleic acid molecules and vectors which encode those peptides or proteins.

Abstract: Described herein is a protein capable of spontaneously forming an isopeptide bond for the development of a peptide tag and binding partner pair wherein the peptide tag and binding partner are capable of covalently binding to each other via an isopeptide bond. Also provided is a method for developing a peptide tag and binding partner pair which are capable of covalently binding to each other based on a protein which is capable of spontaneously forming an isopeptide bond. Additionally provided are peptide tag and binding partner pairs which are obtainable from isopeptide proteins. Further, specifically developed peptide tags and binding partners are described, together with nucleic acid molecules and vectors which encode those peptides or proteins.

Abstract: The present invention provides a mutant streptavidin subunit which comprises one or more amino acid substitutions compared to a wildtype streptavidin subunit at any one of residue positions equivalent to positions (50, 51, 52, 53) and (54) of SEQ ID NO. 2 and wherein amino acid residues at positions equivalent to positions (23, 27, 43, 45, 49, 79, 88, 90, 92, 108, 110) and (128) of SEQ ID NO. 2, in said mutant streptavidin subunit, are wildtype, wherein (i) when said mutant streptavidin subunit unit is comprised in streptavidin, said streptavidin has a lower off rate for biotin or for a biotin conjugate than wildtype streptavidin or (ii) when said mutant streptavidin subunit is in monomelic form said mutant monomelic streptavidin has a lower off rate for biotin or a biotin conjugate than monomelic streptavidin. The invention also encompasses nucleic acid molecules comprising a nucleotide sequence encoding the mutant streptavidin subunit and vectors and cells comprising the nucleic acid.