Patent ID: 6242173
Filing Date: 2001-06-05
Classification: G01N,Y10T

Abstract:
A method of detecting or quantifying a catalytically-active form of a particular serine protease in a biological fluid, comprising the steps of:(a) incubating with the biological fluid:(i) a biotinylated peptidyl halomethyl ketone specific for catalytically-active forms of serine proteases and does not bind the inactive forms, under conditions sufficient for the biotinylated peptidyl halomethyl ketone to be incorporated covalently into the serine proteases, the biotinylated peptidyl halomethyl ketone comprising a peptidyl halomethyl ketone linked to biotin through a spacer group sufficient to make the biotin available for avidin binding after covalent incorporation of the biotinylated peptidyl halomethyl ketone into a serine protease; and(ii) an antibody specific for the particular serine protease under conditions which allow the antibody to bind the serine protease; and(b) determining the amount of serine protease bound by both the peptidyl halomethyl ketone and the antibody as indicative of the presence or amount of the catalytically-active form of the particular serine protease in the biological fluid.