Method of selecting hla-dp4 ligands and the applications...

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G01N 33/68 (2006.01) C07K 14/47 (2006.01) C12N 15/00 (2006.01)

Patent

CA 2463705

The invention relates to a method of selecting HLA-DP4 ligand molecules comprising the following steps: (i) incubation of purified HLA-DP4 with a tracer having general formula (I) Z¿1?X¿1?X¿2?X¿3?X¿4?X¿5?X¿6?X¿7?X¿8?X¿9?Z¿2? (wherein Z¿1? and Z¿2?, which may be identical or different, are equal to zero or each represent a peptide with between 1 and 100 amino acids; X¿6? represents an aromatic or hydrophobic amino acid or a cysteine; and X¿1? represents an aromatic or hydrophobic amino acid and/or X¿9? represents an aromatic or hydrophobic amino acid or C, D, Q, S, T or E; and X¿2?,X¿3?,X¿4?,X¿5?,X¿7? and X¿8? each represent an amino acid) in the presence of different concentrations of molecule(s) to be tested; (ii) separation of the different complexes formed; (iii) detection of HLA-DP4/peptide tracer complexes; and (iv) selection of ligand molecules having a binding capacity IC¿50? < 1000 nM. The invention also relates to the applications of same.

Procédé de sélection de molécules ligand d'HLA-DP4 comprenant les étapes suivantes: (i) incubation d'HLA-DP4 purifiée avec un traceur de formule générale (I) Z1X1X2X3X4X5X6X7X8X9Z2, dans laquelle: Z1 et Z2, identiques ou différents, sont nuls ou représentent chacun un peptide de 1 à 100 acides aminés, X6 représente un acide aminé aromatique ou hydrophobe ou bien une cystéine, et X1 représente un acide aminé aromatique ou hydrophobe et/ou X9 représente un acide aminé aromatique ou hydrophobe, ou bien C, D, Q, S, T ou E, et X2,X3,X4,X5,X7 et X8 représentent chacun un acide aminé, en présence de différentes concentrations de molécule(s) à tester, (ii) séparation des différents complexes formés, (iii) révélation des complexes HLA-DP4/peptide traceur, et (iv) sélection des molécules ligand qui présentent une activité de liaison IC50 < 1000 nM, et ses applications.

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