Methods and compositions for the generation and use of conformation-specific antibodies

Abstract

The present invention features methods and compositions for the generation and use of conformation-specific antibodies or fragments thereof.

What is claimed is: 
     
         1 . An isolated conformation-specific antibody or antigen-binding fragment thereof that specifically binds to a Xaa-Pro motif of a polypeptide, wherein the peptidyl-prolyl bond of said Xaa-Pro motif is in a cis conformation or a trans conformation and wherein Xaa is any amino acid residue. 
     
     
         2 .- 11 . (canceled) 
     
     
         12 . The isolated conformation-specific antibody or antigen-binding fragment thereof of  claim 1  produced by a process which comprises the steps of:
 (i) providing an antibody library; 
 (ii) contacting said antibody library with a polypeptide comprising a Xaa-Pro motif; 
 (iii) determining binding of an antibody from said antibody library to said Xaa-Pro motif of said polypeptide, wherein said antibody specifically binds to the cis or trans conformation of said Xaa-Pro motif of said polypeptide; and 
 (iv) isolating said antibody, wherein said antibody is a conformation-specific antibody. 
 
     
     
         13 .- 23 . (canceled) 
     
     
         24 . A method of generating the confirmation-specific antibody or antigen-binding fragment thereof of  claim 1 , said method comprising:
 (i) administering a proline-analog-containing antigenic peptide to a host animal, said antigenic peptide comprising a Xaa-Pro motif;   (ii) isolating antisera containing said antibody or fragment thereof produced in said animal; and   (iii) purifying said confirmation-specific antibody or fragment thereof from said antisera.   
     
     
         25 .- 41 . (canceled) 
     
     
         42 . A method of purifying the conformation-specific antibody or antigen-binding fragment thereof of  claim 1 , said method comprising:
 (i) adsorbing a proline-analog-containing antigenic peptide to a support, said antigenic peptide comprising a Xaa-Pro motif, wherein the peptidyl-prolyl bond of said Xaa-Pro motif is in a cis conformation;   (ii) contacting said antigenic peptide on said support with antisera containing said antibody or fragment thereof produced in a host animal, wherein said contacting is under conditions allowing said conformation-specific antibody or fragment thereof to specifically bind to said antigenic peptide; and   (iii) eluting said conformation-specific antibody from said antigenic peptide adsorbed to said support.   
     
     
         43 .- 53 . (canceled) 
     
     
         54 . A method of diagnosing a subject with a disorder, wherein said disorder is associated with a deregulation of PPlase activity, said method comprising:
 (A)
 (i) contacting a sample from said subject with a conformation-specific antibody or antigen-binding fragment thereof that specifically binds to a Xaa-Pro motif of a polypeptide present in said sample, wherein the peptidyl-prolyl bond of said Xaa-Pro motif is in a cis conformation and wherein said Xaa is any amino acid residue; 
 (ii) quantitating the amount of said conformation-specific antibody or antigen-binding fragment thereof bound to said Xaa-Pro motif in said cis conformation; and 
 (iii) comparing the amount of said Xaa-Pro motif in said cis conformation in said sample to the amount of said Xaa-Pro motif in said cis conformation found in subjects diagnosed with said disorder or subjects not diagnosed with said disorder, 
 wherein an increase in the amount of said Xaa-Pro motif in said cis conformation in said subject, in comparison to the amount of said Xaa-Pro motif in said cis conformation in said subjects not diagnosed with said disorder, indicates that said subject may be diagnosed with said disorder: 
   or said method comprising:   (B)
 (i) contacting a sample from said subject with a conformation-specific antibody or antigen-binding fragment thereof that specifically binds to a Xaa-Pro motif of a polypeptide present in said sample, wherein the peptidyl-prolyl bond of said Xaa-Pro motif is in a trans conformation and wherein said Xaa is any amino acid residue; 
 (ii) quantitating the amount of said conformation-specific antibody or antigen-binding fragment thereof bound to said Xaa-Pro motif in said trans conformation; and 
 (iii) comparing the amount of said Xaa-Pro motif in said trans conformation in said sample to the amount of said Xaa-Pro motif in said trans conformation found in subjects diagnosed with said disorder or subjects not diagnosed with said disorder, 
 wherein an increase in the amount of said Xaa-Pro motif in said trans conformation in said subject, in comparison to the amount of said Xaa-Pro motif in said trans conformation in said subjects not diagnosed with said disorder, indicates that said subject may be diagnosed with said disorder. 
   
     
     
         55 .- 65 . (canceled) 
     
     
         66 . A method of treating a subject with a disorder, wherein said disorder is associated with a deregulation of PPlase activity, said method comprising administering to said subject a pharmaceutical composition comprising the conformation-specific antibody or antigen-binding fragment thereof of  claim 1 , wherein said conformation-specific antibody is present in an amount sufficient to treat said disorder. 
     
     
         67 . The method of  claim 66 , wherein said Xaa is serine or threonine. 
     
     
         68 . The method of  claim 66 , wherein said Xaa is phosphorylated. 
     
     
         69 . The method of  claim 66 , wherein said Xaa is not phosphorylated. 
     
     
         70 . The method of  claim 66 , wherein said PPlase is Pin1. 
     
     
         71 . The method of  claim 66 , wherein said method further comprises administering an additional therapeutic agent. 
     
     
         72 . The method of  claim 71 , wherein said additional therapeutic agent is a chemotherapeutic agent. 
     
     
         73 . The method of  claim 66 , wherein said disorder is a cell proliferation disorder or a neurological disorder. 
     
     
         74 . The method of  claim 73 , wherein said cell proliferation disorder is cancer. 
     
     
         75 . The method of  claim 73 , wherein said neurological disorder is Alzheimer's disease. 
     
     
         76 . The method of  claim 66 , wherein said disorder is asthma or a microbial infection. 
     
     
         77 . (canceled) 
     
     
         78 . The method of  claim 66 , wherein said disorder is aging or an aging-related disorder. 
     
     
         79 . An antigenic peptide comprising a Xaa-Pro motif that specifically binds to an antibody or antigen-binding fragment thereof, wherein said Xaa is any amino acid residue and wherein:
 said antibody or antigen-binding fragment thereof binds to the cis conformation of said Xaa-Pro motif of said antigenic peptide with at least 10- to 100-fold greater affinity than to the trans conformation of said Xaa-Pro motif of said polypeptide; or   said antibody or antigen-binding fragment thereof binds to the trans conformation of said Xaa-Pro motif of said antigenic peptide with at least 10- to 100-fold greater affinity than to the cis conformation of said Xaa-Pro motif of said polypeptide.   
     
     
         80 .- 91 . (canceled) 
     
     
         92 . A vaccine composition to induce passive immunity against a disorder associated with a deregulation of PPlase activity, said vaccine composition comprising the isolated antibody or antigen-binding fragment thereof of  claim 1 . 
     
     
         93 .- 99 . (canceled) 
     
     
         100 . A method for providing passive immunity against a disorder associated with a deregulation of PPlase activity in a subject, said method comprising administering to said subject the vaccine composition of  claim 92 .