Plasma protein-binding ligands

Abstract

The invention provides an isolated or purified peptide that binds at least one plasma protein. In one embodiment, the isolated or purified peptide binds to fibrinogen, comprises no more than 10 amino acids, and comprises an amino acid sequence Xaa1-Xaa2-Xaa3-Xaa4-Xaa5, an amino acid sequence Gly-Xaa6-Arg-Xaa7, or an amino acid sequence selected from specific amino acid sequences provided herein. Alternatively, the isolated or purified protein binds to alpha1 proteinase inhibitor and/or a protein complex comprising Apo-A1 lipoprotein and paraoxonase. The peptide comprises no more than 10 amino acids and comprises an amino acid sequence Xaa8-Xaa8-Xaa1-His-Xaa1-Xaa3, and amino acid sequence His-Xaa8-Xaa9-Xaa1-Xaa10-Xaa2, or an amino acid sequence selected from specific amino acid sequences provided herein. In addition, the invention provides isolated or purified peptide that binds to von Willebrand Factor. The peptide comprises an amino acid sequence Xaa4-Xaa5-Xaa5, an amino acid sequence Tyr-Leu-Xaa11-Xaa4-Xaa12-Thr, or an amino acid sequence selected from specific amino acid sequences provided herein.

1 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence Xaa 1 -Xaa 2 -Xaa 3 -Xaa 4 -Xaa 5  (SEQ ID NO: 1), wherein
 Xaa 1  is a hydrophobic amino acid,   Xaa 2  is a basic amino acid,   Xaa 3  is a polar amino acid with a side chain comprising an amide,   Xaa 4  is a hydrophobic amino acid or an aromatic amino acid, and   Xaa 5  is an acidic amino acid,   and wherein the peptide binds to fibrinogen.   
     
     
         2 . The isolated or purified peptide of  claim 1 , wherein the amino acid sequence comprises an N-terminal amino acid that is a D-amino acid. 
     
     
         3 . The isolated or purified peptide of  claim 2 , wherein Xaa 1  is Ala, Xaa 2  is Arg, Xaa 3  is Asn or Gln, and Xaa 5  is Asp. 
     
     
         4 . The isolated or purified peptide of  claim 3 , wherein the peptide consists essentially of SEQ ID NO: 20 or SEQ ID NO: 21. 
     
     
         5 . A composition comprising the peptide of  claim 1  and a carrier. 
     
     
         6 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence Gly-Xaa 6 -Arg-Xaa 7  (SEQ ID NO: 2), wherein
 Xaa 6  is Pro or Gln, and   Xaa 7  is any amino acid except Pro,   and wherein the peptide binds to fibrinogen.   
     
     
         7 . The isolated or purified peptide of  claim 6 , wherein Xaa 6  is Pro. 
     
     
         8 . The isolated or purified peptide of  claim 6 , wherein the peptide consists essentially of an amino acid sequence selected from the group consisting of SEQ ID NOs: 16, 22-27, and 29-32. 
     
     
         9 . A composition comprising the peptide of  claim 6  and a carrier. 
     
     
         10 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 7-15, 17-19, and 28, wherein the peptide binds to fibrinogen. 
     
     
         11 . The isolated or purified peptide of  claim 10 , wherein the amino acid sequence is selected from the group consisting of SEQ ID NOs: 17-19 and comprises an N-terminal amino acid that is a D-amino acid. 
     
     
         12 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying fibrinogen in a sample, which method comprises (a) contacting a sample comprising fibrinogen with the peptide of  claim 1  to form a fibrinogen-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the fibrinogen-peptide complex. 
     
     
         13 . The method of  claim 12 , wherein the peptide is attached to a support. 
     
     
         14 . The method of  claim 13 , wherein the support is a chromatography resin or a membrane. 
     
     
         15 . The method of  claim 12 , wherein the peptide consists essentially of an amino acid sequence of SEQ ID NO: 20 or SEQ ID NO: 21. 
     
     
         16 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying fibrinogen in a sample, which method comprises (a) contacting a sample comprising fibrinogen with the peptide of  claim 6  to form a fibrinogen-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the fibrinogen-peptide complex. 
     
     
         17 . The method of  claim 16 , wherein the peptide is attached to a support. 
     
     
         18 . The method of  claim 17 , wherein the support is a chromatography resin or a membrane. 
     
     
         19 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying of removing fibrinogen in a sample, which method comprises (a) contacting a sample comprising fibrinogen with the peptide of  claim 10  to form a fibrinogen-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the fibrinogen-peptide complex. 
     
     
         20 . The method of  claim 19 , wherein the peptide is attached to a support. 
     
     
         21 . The method of  claim 20 , wherein the support is a chromatography resin or a membrane. 
     
     
         22 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence Xaa 8 -Xaa 8 -Xaa 1 -His-Xaa 1 -Xaa 3  (SEQ ID NO: 3), wherein
 Xaa 1  is a hydrophobic amino acid,   Xaa 3  is a polar amino acid with a side chain comprising an amide, and   Xaa 8  is an aromatic amino acid, and   wherein the peptide binds to α1 proteinase inhibitor (API) and/or a protein complex comprising paraoxonase and Apo-A1 lipoprotein.   
     
     
         23 . The isolated or purified peptide of  claim 22 , wherein Xaa 8  at position 1 is Trp, Tyr, 1-naphthylalanine (na1′), or 2-naphthylalanine (na2′), and Xaa 3  at position 6 is Asn or Gln. 
     
     
         24 . The isolated or purified peptide of  claim 23 , wherein Xaa 8  at position 2 is Trp, Tyr, na1′, or na2′, Xaa 1  at position 3 is Leu, and Xaa 1  at position 5 is Ile. 
     
     
         25 . The isolated or purified peptide of  claim 22 , wherein amino acid sequence comprises an N-terminal amino acid that is a D-amino acid. 
     
     
         26 . A composition comprising the peptide of  claim 22  and a carrier. 
     
     
         27 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence His-Xaa 8 -Xaa 9 -Xaa 1 -Xaa 10 -Xaa 2  (SEQ ID NO: 4), wherein
 Xaa 1  is a hydrophobic amino acid,   Xaa 2  is a basic amino acid,   Xaa 8  is an aromatic amino acid,   Xaa 9  is an acidic amino acid or a hydrophobic amino acid, and   Xaa 10  is a hydrophobic amino acid or His,   and wherein the peptide binds to API.   
     
     
         28 . The isolated or purified peptide of  claim 27 , wherein the peptide consists essentially of L-amino acids. 
     
     
         29 . The isolated or purified peptide of  claim 27 , wherein the peptide consists essentially of an amino acid sequence of SEQ ID NO: 46 or SEQ ID NO: 47. 
     
     
         30 . A composition comprising the peptide of  claim 27  and a carrier. 
     
     
         31 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 36, 38, 39, 45, 48, 54, and 55, wherein the peptide binds to API. 
     
     
         32 . The isolated or purified peptide of  claim 31 , wherein the amino acid sequence comprises an N-terminal amino acid that is a D-amino acid. 
     
     
         33 . A composition comprising the peptide of  claim 31  and a carrier. 
     
     
         34 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 59-62, wherein the peptide binds to a protein complex comprising Apo-A1 lipoprotein and paraoxonase. 
     
     
         35 . A composition comprising the peptide of  claim 34  and a carrier. 
     
     
         36 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying API and/or a protein complex comprising Apo-A1 lipoprotein and paraoxonase in a sample, which method comprises (a) contacting a sample comprising API and/or a protein complex comprising Apo-A1 lipoprotein and paraoxonase with the peptide of  claim 22  to form an API-peptide complex or a complex comprising the peptide and Apo-A1 lipoprotein and/or paraoxonase, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the API-peptide complex and/or the complex comprising the peptide and Apo-A1 lipoprotein and/or paraoxonase. 
     
     
         37 . The method of  claim 36 , wherein the peptide is attached to a support. 
     
     
         38 . The method of  claim 37 , wherein the support is a chromatography resin or a membrane. 
     
     
         39 . The method of  claim 36 , wherein the peptide consists essentially of an amino acid sequence selected from the group consisting of SEQ ID NOs: 34, 39, 40, 50, 54 and 55. 
     
     
         40 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying API in a sample, which method comprises (a) contacting a sample comprising API with the peptide of  claim 27  to form an API-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the API-peptide complex. 
     
     
         41 . The method of  claim 40 , wherein the peptide is attached to a support. 
     
     
         42 . The method of  claim 41 , wherein the support is a chromatography resin or a membrane. 
     
     
         43 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying API in a sample, which method comprises (a) contacting a sample comprising API with the peptide of  claim 31  to form an API-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the API-pep tide complex. 
     
     
         44 . The method of  claim 43 , wherein the peptide is attached to a support. 
     
     
         45 . The method of  claim 44 , wherein the support is a chromatography resin or a membrane. 
     
     
         46 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying a protein complex comprising Apo-A1 lipoprotein and paraoxonase in a sample, which method comprises (a) contacting a sample comprising a protein complex comprising Apo-A1 lipoprotein and paraoxonase with the peptide of  claim 34  to form a complex comprising Apo-A1 lipoprotein, paraoxonase, and the peptide of  claim 34 , and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the complex. 
     
     
         47 . The method of  claim 46 , wherein the peptide is attached to a support. 
     
     
         48 . The method of  claim 47 , wherein the support is a chromatography resin or a membrane. 
     
     
         49 . An isolated or purified peptide comprising no more than 6 amino acids and comprising an amino acid sequence Xaa 4 -Xaa 5 -Xaa 5 , wherein
 Xaa 4  is a hydrophobic amino acid or an aromatic amino acid, and   Xaa 5  is an acidic amino acid, and   wherein the peptide does not comprise His, Arg, or Lys, and binds to von Willebrand Factor (vWF).   
     
     
         50 . The isolated or purified peptide of  claim 49 , wherein Xaa 5  is Asp or Glu. 
     
     
         51 . The isolated or purified peptide of  claim 49 , wherein the peptide consists essentially of an amino acid sequence selected from the group consisting of SEQ ID NOs: 64, 65, 68-73 and 111. 
     
     
         52 . A composition comprising the peptide of  claim 49  and a carrier. 
     
     
         53 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence Tyr-Leu-Xaa 1 -Xaa 4 -Xaa 12 -Thr, wherein
 Xaa 4  is a hydrophobic amino acid or an aromatic amino acid,   Xaa 1  is an aromatic amino acid or His, and   Xaa 12  is a hydrophobic amino acid or a polar amino acid, and   wherein the peptide binds to vWF.   
     
     
         54 . The isolated or purified peptide of  claim 53 , wherein the Xaa 1  is His or Tyr, Xaa 4  is Tyr or Ala, and Xaa 12  is Gln or Leu. 
     
     
         55 . An isolated or purified peptide comprising no more than 10 amino acids and comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 67, 76, and 77, wherein the peptide binds to vWF. 
     
     
         56 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying vWF in a sample, which method comprises (a) contacting a sample comprising vWF with the peptide of  claim 49  to form a vWF-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the vWF-peptide complex. 
     
     
         57 . The method of  claim 56 , wherein the peptide is attached to a support. 
     
     
         58 . The method of  claim 57 , wherein the support is a chromatography resin or a membrane. 
     
     
         59 . The method of  claim 57 , wherein Factor VIII is bound to vWF, and the Factor VIII is co-purified with the vWF. 
     
     
         60 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying vWF in a sample, which method comprises (a) contacting a sample comprising vWF with the peptide of  claim 53  to form a vWF-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the vWF-peptide complex. 
     
     
         61 . The method of  claim 60 , wherein the peptide is attached to a support. 
     
     
         62 . The method of  claim 61 , wherein the support is a chromatography resin or a membrane. 
     
     
         63 . The method of  claim 61 , wherein Factor VIII is bound to vWF, and the Factor VIII is co-purified with the vWF. 
     
     
         64 . A method of separating, isolating, purifying, characterizing, identifying, or quantifying vWF in a sample, which method comprises (a) contacting a sample comprising vWF with the peptide of  claim 55  to form a vWF-peptide complex, and (b) separating, isolating, purifying, characterizing, identifying, or quantifying the vWF-peptide complex. 
     
     
         65 . The method of  claim 64 , wherein the peptide is attached to a support. 
     
     
         66 . The method of  claim 65 , wherein the support is a chromatography resin or a membrane. 
     
     
         67 . The method of  claim 64 , wherein Factor VIII is bound to vWF, and the Factor VIII is co-purified with the vWF.