Claims
- 1. A method for selecting an artificial bacterial receptor structure, the method comprising:producing an expression library that produces a protein comprising an artificial bacterial receptor structure, wherein the amino acid sequence of the artificial bacterial receptor structure corresponds to that of a natural bacterial receptor having at least one surface-exposed amino acid residue substituted by another amino acid residue, wherein the basic structure and stability of the natural bacterial receptor is not lost, wherein the artificial bacterial receptor structure lacks an interaction capacity of the natural bacterial receptor, and wherein the artificial bacterial receptor structure binds to an interaction partner to which the natural bacterial receptor does not bind; screening the expression library with a putative binding partner that is not bound by the natural bacterial receptor; detecting specific binding of the artificial bacterial receptor structure to the putative binding partner; and selecting a DNA from the expression library that encodes the artificial bacterial receptor structure that specifically binds to the putative binding partner.
- 2. The method of claim 1, wherein the protein comprises a phage-coat protein.
- 3. The method of claim 1, wherein the natural bacterial receptor is a receptor in a bacterial species selected from the group consisting of Staphylococcus aureus, Streptococcus pyogenes (group A), Streptococcus groups C,G,L, bovine group G streptococci, Streptococcus zooepidemicus (group C), Streptococcus zooepidemicus S212, streptococci groups A,C,G, Peptostreptococcus magnus, and Streptococcus agalactiae (group B).
- 4. The method of claim 1, wherein the natural bacterial receptor is staphylococcal protein A or streptococcal protein G.
- 5. The method of claim 1, wherein the natural bacterial receptor is selected from the group consisting of: Fc receptor IgG type I, type II, type III, type IV, type V, and type VI; fibronectin receptor; M protein; plasmin receptor; collagen receptor; fibrinogen receptor; protein L; protein H; protein B; and protein Arp.
- 6. The method of claim 1, wherein the natural bacterial receptor is the Fc receptor IgG type I of staphylococcal protein A or the serum albumin receptor of streptococcal protein G.
- 7. The method of claim 1, wherein at most about 50% of the amino acid residues of the natural bacterial receptor have been substituted by other amino acid residues.
- 8. The method of claim 7, wherein at most about 25% of the amino acid residues of the natural bacterial receptor have been substituted by other amino acid residues.
- 9. The method of claim 8, wherein the natural bacterial receptor is staphylococcal protein A or streptococcal protein G.
- 10. The method of claim 9, wherein the interaction partner is selected from the group consisting of IgF-I, IGF-II, hGH, Factor VIII, insulin, apolipoprotein, and their respective receptors.
- 11. The method of claim 8, wherein the natural bacterial receptor is selected from the group consisting of: Fc receptor IgG type I, type II, type III, type IV, type V, and type VI; fibronectin receptor; M protein; plasmin receptor; collagen receptor; fibrinogen receptor; protein L; protein H; protein B; and protein Arp.
- 12. The method of claim 8, wherein the natural bacterial receptor is the Fc receptor IgG type I of staphylococcal protein A or the serum albumin receptor of streptococcal protein G.
- 13. The method of claim 1, wherein only surface-exposed amino acid residues of the natural bacterial receptor have been substituted.
- 14. The method of claim 1, wherein the interaction partner is selected from the group consisting of a protein, lipid, carbohydrate, and inorganic substance.
- 15. The method of claim 14, wherein the interaction partner is a carbohydrate.
- 16. The method of claim 14, wherein the interaction partner is selected from the group consisting of IgF-I, IGF-II, hGH, Factor VIII, insulin, apolipoprotein, and their respective receptors.
- 17. The method of claim 14, wherein the interaction partner is selected from the group consisting of a viral coat protein, bacterial antigen, biotin, and cell marker.
- 18. The method of claim 14, wherein the interaction partner is an antibody fragment.
- 19. The method of claim 14, wherein the interaction partner is an organic ligand.
- 20. The method of claim 1, wherein the natural bacterial receptor comprises an ααα domain structure.
- 21. The method of claim 1, wherein the natural bacterial receptor comprises a ββαββ domain structure.
- 22. The method of claim 1, wherein the artificial bacterial receptor structure is produced by site-specific mutagenesis of at least two codons that encode surface exposed amino acid residues of the natural bacterial receptor.
- 23. The method of claim 1, wherein the expression library comprises phage particles that express on their surface a repertoire of artificial bacterial receptor structures, wherein the artificial bacterial receptor structures are produced by the expression of fusions comprising phage-coat proteins fused to artificial bacterial receptor structures, and wherein the expression library comprising phage particles is panned with the putative binding partner to identify a phage particle that expresses on its surface the artificial bacterial receptor structure that specifically binds to the putative binding partner.
- 24. The method of claim 23, further comprising isolating the phage particle that specifically binds to the putative binding partner.
- 25. The method of claim 1, wherein the expression library comprises bacterial cells that express on their surface artificial bacterial receptor structures, wherein the artificial bacterial receptor structures are fused to cell-wall anchoring domains which results in stable surface exposure of the resultant fusion proteins on the surface of the bacterial cells, and wherein the expression library comprising bacterial cells is panned with the putative binding partner to identify a bacterial clone that expresses the artificial bacterial receptor structures that specifically binds to the putative binding partner.
- 26. The method of claim 25, further comprising isolating the bacterial clone from the panned expression library comprising bacterial cells.
- 27. The method of claim 1, wherein the expression library comprises bacterial cells comprising the DNA that encodes the artificial bacterial receptor structure fused to a repressor protein that has affinity for an operator, and wherein the bacterial cells are isolated based on a repressor-operator interaction.
- 28. The method of claim 1, wherein the natural bacterial receptor is the Z IgG binding domain or the B2A3 serum albumin binding domain.
- 29. The method of claim 1, wherein the natural bacterial receptor is the Ig receptor of streptococcal protein G.
- 30. The method of claim 1, wherein the natural bacterial receptor is the C1 IgG binding domain.
- 31. The method of claim 1, wherein all of the amino acid residues of the natural bacterial receptor that are involved in the binding of the natural bacterial receptor with its native binding partner have been substituted by other amino acid residues.
- 32. The method of claim 1, wherein the interaction partner is a carbohydrate comprising blood group determinants or pathogenic specific oligosaccharides.
- 33. The method of claim 1, wherein the interaction partner is CD34 or CD4.
- 34. The method of claim 1, wherein the interaction partner is an antibody fragment selected from the group consisting of Fv, ScFv, Fab, and Fc.
- 35. The method of claim 1, wherein the natural bacterial receptor is staphylococcal protein A, and wherein the artificial bacterial receptor structure comprises substitutions of amino acid residues 9, 11, 14, 27, and 35 of the amino acid sequence of SEQ ID NO:16.
- 36. The method of claim 35, wherein the substitutions result in a glutamic acid residue at position 9, an aspartic acid at position 11, an aspartic acid at position 14, an alanine at position 27, and either a glutamic acid or alanine at position 35 of the amino acid sequence of SEQ ID NO:16.
- 37. The method of claim 1, wherein the natural bacterial receptor is staphylococcal protein A, and wherein the artificial bacterial receptor structure comprises substitutions such that at least two of residues 9, 10, 11, 13, 14, 17, 28, 32, 35, 18, 24, 25, and 27 in the Z Ig binding domain of SEQ ID NO:16 are modified.
- 38. The method of claim 1, wherein the natural bacterial receptor is staphylococcal protein A, and wherein the artificial bacterial receptor structure comprises an amino acid sequence selected from SEQ ID Nos:17-47.
Priority Claims (1)
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9400088 |
Jan 1994 |
SE |
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CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a divisional of U.S. application Ser. No. 09/082,468, filed, May 21, 1998, now U.S. Pat. No. 6,534,628, which is a continuation of U.S. application Ser. No. 08/669,360 filed Aug. 15, 1996, now U.S. Pat. No. 5,831,012, which is a section 371 of PCT/SE95/00034, filed Jan. 16, 1995, which claims priority to Swedish Application No. 9400088-2 filed, Jan. 14, 1994.
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Number |
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WO 9220805 |
Nov 1992 |
WO |
WO 9519374 |
Jul 1995 |
WO |
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Continuations (1)
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08/669360 |
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09/082468 |
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