Patent ID: 11959913
Assignee: ADIENNE S.A.
Field: Measurement (Instruments)
Classification: CPC G  C | IPC C  G

Claim 0:
1. A method for the in vitro determination of the potency of an anti-CD26 ligand comprising the following steps:
a) incubating at 37° C. or at room temperature a population of human T lymphocytes expressing the CD26 receptor in a percentage higher than 75% with an anti-CD26 ligand at a concentration ranging from 0.001 μg/ml to 150 μg/ml;
b) contacting said human T lymphocytes with an anti-CD26 antibody conjugated to a fluorochrome; wherein the anti-CD26 antibody recognizes a different CD26 epitope from the epitope recognized by the anti-CD26 ligand used in step a);
c) determining the mean fluorescence intensity (MFI) of CD26 measured for the sample of cells treated with the anti-CD26 ligand (MFIT) and the MFI value of untreated cells (MFINT) by means of cytofluorimetric analysis;
d) evaluating the internalization percentage of the CD26 receptor (% int CD26) or RFI calculated according to the following formula:, %
   ⁢
      
   int
   ⁢
      
   C
   ⁢
   D
   ⁢
   26
  
  =
  
   100
   -
   
    (
    
     
      
       M
       ⁢
       F
       ⁢
       
        I
        T
       
      
      
       M
       ⁢
       F
       ⁢
       
        I
        NT
       
      
     
     ×
     100
    
    ), wherein if the value of % intCD26 is:
less than 20% it indicates a low potency of the anti-CD26 ligand;
ranging from 20% to 30% it indicates a medium potency of the anti-CD26 ligand; and
higher than 30% indicates a high potency of the anti-CD26 ligand.