C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/37 (2006.01) C07K 4/00 (2006.01) C07K 5/083 (2006.01) G01N 33/573 (2006.01) A61K 38/00 (2006.01)
Patent
CA 2198352
The present invention relates to a method of detecting the presence of an active protease in a sample. The method involves the following steps: (1) providing a solid support to which is attached a ligand reactive with a protease; (2) contacting the solid support with a sample suspected to contain the protease; (3) washing the solid support to remove material not bound to the ligand; (4) contacting the washed solid support with a solution containing a peptide ester substrate which is hydrolysed for the protease; and (5) detecting protons produced by hydrolysis of the peptide ester substrate. In addition, the present invention provides a prostate specific antigen substrate, the substrate being of the general formula: A-B-Tyrosine-ester in which A is acetyl or carbobenzoxy and B is 2 to 6 amino acids.
L'invention concerne un procédé qui permet de détecter la présence d'une protéase active dans un échantillon. Ce procédé consiste à: (1) prendre un support solide auquel est fixé un ligand pouvant réagir avec une protéase; (2) mettre ce support en contact avec un échantillon soupçonné de contenir la protéase; (3) laver le support solide pour éliminer les éléments non liés au ligand; (4) mettre le support solide lavé en contact avec une solution contenant un substrat à base de peptide-ester qui est hydrolysé en fonction de cette protéase; et (5) détecter des protons produits par l'hydrolyse de ce substrat à base de peptide-ester. L'invention porte aussi sur un substrat contenant un antigène spécifique de la prostate, ce substrat correspondant à la formule générale: A-B-tyrosine-ester, où A représente acétyle ou carbobenzoxy, et B représente 2 à 6 acides aminés.
Whittaker Robert George
Yu Yan
Commonwealth Scientific And Industrial Research Organisation
Swabey Ogilvy Renault
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