US2007292947A1PendingUtilityA1

Novgel methods for displaying (poly)peptides/proteins on bacteriophage particles via disulfide bonds

58
Assignee: MORPHOSYS AGPriority: Jul 20, 1999Filed: Feb 28, 2007Published: Dec 20, 2007
Est. expiryJul 20, 2019(expired)· nominal 20-yr term from priority
Inventors:Corinna Löhning
C12N 15/1037C40B 40/02C07K 14/005C12N 2795/10022
58
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Claims

Abstract

The present invention relates to methods for displaying (poly)peptides/proteins on the surface of bacteriophage particles by attaching the (poly)peptide/proteins via disulfide bonds.

Claims

exact text as granted — not AI-modified
1 - 31 . (canceled)  
     
     
         32 . An isolated host cell comprising: 
 (a) a first vector comprising a nucleic acid sequence encoding a variant of a wild type coat protein of a bacteriophage, wherein said variant comprises: 
 (aa) one or more parts of said wild type coat protein of a bacteriophage, wherein one of said parts comprises at least that part which causes or allows the incorporation of said coat protein into the phage coat; and,  
 (ab) between one and six additional amino acid residues not present at the corresponding amino acid positions in a wild type coat protein of a bacteriophage, wherein one of said additional amino acid residues is a cysteine residue, and  
   (b) a second vector comprising one or more nucleic acid sequences encoding a (poly)peptide/protein comprising a cysteine residue.    
     
     
         33 . The isolated host cell of  claim 32 , wherein said bacteriophage is a filamentous bacteriophage.  
     
     
         34 . An isolated host cell comprising: 
 (a) a first vector comprising a nucleic acid sequence encoding a wild type coat protein or a truncated portion thereof of a bacteriophage, wherein said wild type coat protein or said truncated portion thereof comprises: 
 (aa) one or more parts of said wild type coat protein or said truncated variant thereof of a bacteriophage, wherein one of said parts comprises at least that part which causes or allows the incorporation of said coat protein into the phage coat; and,  
 (ab) a cysteine residue which is present at a corresponding amino acid position in a wild type coat protein or a truncated portion thereof of a bacteriophage, and  
   (b) a second vector comprising one or more nucleic acid sequences encoding a (poly)peptide/protein comprising a cysteine residue.    
     
     
         35 . The isolated host cell of  claim 34 , wherein said bacteriophage is a filamentous bacteriophage.  
     
     
         36 . The isolated host cell of  claim 32 , wherein said nucleic acid sequence encoding said variant of said wild type coat protein further encodes: 
 (c) one or more peptide sequences for purification and/or detection purposes, wherein said one or more peptide sequences are fused to said variant of said wild type coat protein.    
     
     
         37 . The isolated host cell of  claim 34 , wherein said nucleic acid sequence encoding said wild type coat protein or said truncated portion thereof further encodes: 
 (c) one or more peptide sequences for purification and/or detection purposes, wherein said one or more peptide sequences are fused to said variant of said wild type coat protein.    
     
     
         38 . The isolated host cell of  claim 32 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         39 . The isolated host cell of  claim 34 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         40 . The isolated host cell of  claim 32 , wherein said (poly)peptide/protein comprises an immunoglobulin or a functional fragment thereof.  
     
     
         41 . The isolated host cell of  claim 34 , wherein said (poly)peptide/protein comprises an immunoglobulin or a functional fragment thereof.  
     
     
         42 . A vector comprising 
 (a) a nucleic acid sequence encoding a variant of a wild type coat protein of a bacteriophage, wherein said variant comprises: 
 (aa) one or more parts of said wild type coat protein of a bacteriophage, wherein one of said parts comprises at least that part which causes or allows the incorporation of said coat protein into the phage coat; and  
 (ab) between one and six additional amino acid residues not present at the corresponding amino acid positions in a wild type coat protein of a bacteriophage, wherein one of said additional amino acid residues is a cysteine residue; and  
   (b) one or more nucleic acid sequences encoding a (poly)peptide/protein comprising a cysteine residue.    
     
     
         43 . The vector of  claim 42 , wherein said bacteriophage is a filamentous bacteriophage.  
     
     
         44 . The vector of  claim 42 , wherein the expression product of said nucleic acid sequence encoding said variant of a wild type coat protein and the expression product of said nucleic acid sequences encoding a (poly)peptide/protein do not form a genetic fusion protein.  
     
     
         45 . The vector of  claim 42 , wherein no interaction domain for interaction with a second domain present in the (poly)peptide/protein has been recombinantly fused to said coat protein.  
     
     
         46 . The vector of  claim 42 , wherein said (poly)peptide/protein comprises an immunoglobulin or a functional fragment thereof.  
     
     
         47 . The vector of  claim 44 , wherein said (poly)peptide/protein comprises an immunoglobulin or a functional fragment thereof.  
     
     
         48 . The vector of  claim 45 , wherein said (poly)peptide/protein comprises an immunoglobulin or a functional fragment thereof.  
     
     
         49 . An isolated host cell comprising a vector of  claim 42 .  
     
     
         50 . An isolated host cell comprising a vector of  claim 43 .  
     
     
         51 . An isolated host cell comprising a vector of  claim 44 .  
     
     
         52 . An isolated host cell comprising a vector of  claim 45 .  
     
     
         53 . An isolated host cell comprising a vector of  claim 46 .  
     
     
         54 . An isolated host cell comprising a vector of  claim 47 .  
     
     
         55 . An isolated host cell comprising a vector of  claim 48 .  
     
     
         56 . The isolated host cell of  claim 49 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         57 . The isolated host cell of  claim 50 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         58 . The isolated host cell of  claim 51 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         59 . The isolated host cell of  claim 52 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         60 . The isolated host cell of  claim 53 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         61 . The isolated host cell of  claim 54 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         62 . The isolated host cell of  claim 55 , wherein said host cell is a bacterial, fungal, plant, insect or mammalian host cell.  
     
     
         63 . The isolated host cell of  claim 49 , wherein said bacteriophage is a filamentous bacteriophage.  
     
     
         64 . An isolated nucleic acid sequence encoding 
 (a) a variant of a wild type coat protein of a bacteriophage, wherein said variant comprises: 
 (aa) one or more parts of said wild type coat protein of a bacteriophage, wherein one of said parts comprises at least that part which causes or allows the incorporation of said coat protein into the phage coat; and  
 (ab) between one and six additional amino acid residues not present at the corresponding amino acid positions in a wild type coat protein of a bacteriophage, wherein one of said additional amino acid residues is a cysteine residue; and  
   (b) a (poly)peptide/protein comprising a cysteine residue.    
     
     
         65 . The isolated nucleic acid of  claim 64 , wherein said (poly)peptide/protein comprising the cysteine residue is selected from the group consisting of immunoglobulins, VH, VL, Fv, scFv, disulfide-linked Fv, Fab and F(ab′)2.  
     
     
         66 . An isolated host cell comprising the nucleic acid sequence of  claim 64 .  
     
     
         67 . An isolated host cell comprising the nucleic acid sequence of  claim 65.

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