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US 11,959,913 B2Granted
Quantitative cellular method for determining the biological activity of an anti-CD26 ligand
AssigneeAdienne S.A.
InventorAntonio Francesco Di Naro
Life of this patent
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Abstract
The present invention relates to a quantitative cellular method for the in vitro determination of the effect of an anti-CD26 ligand, preferably of an anti-CD26 monoclonal antibody, such as begelomab.
Claims12
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.
35 more elements
Dependencies are inferred from the claim text.