Process for detecting PrP using a macrocyclic adjuvant ligand

Abstract

A process for detecting the forms of the prion pathogens responsible for subacute, transmissible, spongiform encephalopathies, including a macrocyclic adjuvant ligand (AML), free or bound to a support, that is added to a biological sample capable of containing PrP sc , the resulting suspension then being reacted with an anti-PrP sc antibody, and the presence of PrP is then detected.

1 - 9 . (canceled)  
   
   
       10 . The process according to claim  9 , wherein the macrocyclic adjuvant ligand corresponds to the general formula (Ia) below:  
     
       
         
         
             
             
         
       
     
     in which 
 each R 2  group, taken independently, is a sulfate group or a phosphate group,  
 R 7  is a (CH 2 )r-Z group in which Z is a COOEt, COOH, CN or NH 2  group,  
 r is a positive integer between 1 and 20,  
 p is a positive integer between 1 and 10, and  
 n is a positive integer between 2 and 10.  
 
   
   
       11 . The process according to claim  9 , wherein the ligand corresponds to formula (Ib) below:  
     
       
         
         
             
             
         
       
     
     in which 
 n is a positive integer between 4 and 8,  
 each R 2  group, taken independently, is a sulfate group or a phosphate group, and  
 R 8  represents a (CH 2 ) t —(CO) s —(NH 2 ) group or a (CH 2 ) t —COOH group in which t is a positive integer between 0 and 6 and s is a positive integer between 0 and 6.  
 
   
   
       12 . The process according to  claim 11 , wherein the ligand is a calyx-arene of formula (Ib) in which each of the two groups R 2  is a sulfate group, n is 4, 6 or 8 and R 8  is a hydrogen atom, a CH 2 COOH, group, a CH 2 CONH 2  group or a CH 2 CH 2 NH 2  group.  
   
   
       13 . The process according to  claim 11 , wherein each of the two R 2  groups is a sulfate group, n is a positive integer equal to 6 and R 7  is a (CH 2 ) 2 -NH 2  group.  
   
   
       14 . The process according to claim  7 , wherein calyx arene is bound to a functionalized support by an NHS or NH 2  function.  
   
   
       15 . A macrocyclic adjuvant ligand grafted onto a functionalized support in which the ligand corresponds to the formula (Ib):  
     
       
         
         
             
             
         
       
     
     in which 
 n is a positive integer between 4 and 8,  
 each R 2  group, taken independently, is a sulfate group or a phosphate group,  
 R 8  represents a (CH 2 ) t —(CO) s —(NH 2 ) group or a (CH 2 ) t —COOH group in which t is a positive integer between 0 and 6 and s is a positive integer between 0 and 6, and the support is a solid support functionalized by an NHS group or an NH 2  group and is a magnetic ball or a microplate.  
 
   
   
       16 . A diagnostic kit for diseases involving PrP sc  comprising a macrocyclic adjuvant ligand of formula (I), as defined in claim  9 , free or bound to a support.  
   
   
       17 . A diagnostic kit for diseases involving PrP sc  comprising a macrocyclic adjuvant ligand of formula (Ia), as defined in  claim 10 , free or bound to a support.  
   
   
       18 . A diagnostic kit for diseases involving PrP sc  comprising a macrocyclic adjuvant ligand of formula (Ib), as defined in  claim 15 , free or bound to a support.  
   
   
       19 . A kit for the immunological dosing of PrP sc  comprising a macrocyclic adjuvant ligand of formula (I), as defined in claim  9 , free or bound to a support.  
   
   
       20 . A kit for the immunological dosing of PrP sc  comprising a macrocyclic adjuvant ligand of formula (Ia), as defined in  claim 10 , free or bound to a support.  
   
   
       21 . A kit for the immunological dosing of PrP sc  comprising a macrocyclic adjuvant ligand of formula (Ib), as defined in  claim 15 , free or bound to a support.  
   
   
       22 . A process for detecting PrP sc , comprising: 
 adding a macrocyclic adjuvant ligand (AML) that is free or linked to a support to a biological sample that may contain PrP sc  to form a suspension;    reacting the suspension with an anti-PrP antibody    subjecting the biological sample to conditions that degrade PrP, either before or after reacting adding the macrocyclic adjuvant ligand (AML); and    identifying the presence of PrP sc ;    wherein said AML is selected from the group consisting of metacyclophanes, cyclodextrins and veratrylenes.