公开(公告)号：US5235039A

公开(公告)日：1993-08-10

申请号：US712828

申请日：1991-06-10

申请人： William F. Heath, Jr. ,  Mei-Huei T. Lai ,  Joseph V. Manetta ,  John R. Sportsman ,  Sau-Chi B. Yan

发明人： William F. Heath, Jr. ,  Mei-Huei T. Lai ,  Joseph V. Manetta ,  John R. Sportsman ,  Sau-Chi B. Yan

IPC分类号： C07K7/08 ,  C07K7/06 ,  C07K7/22 ,  C07K14/16 ,  C07K14/78 ,  C12Q1/34 ,  C12Q1/37 ,  G01N21/78 ,  G01N33/573

CPC分类号： C07K14/005 ,  C07K14/78 ,  C07K7/22 ,  C12Q1/37 ,  G01N33/573 ,  C12N2740/16222 ,  G01N2333/16 ,  G01N2333/78

摘要： An assay method for the rapid determination of hydrolytic enzyme activity in large numbers of samples is provided which comprises bonding a resin-binding compound, such as biotin, to one side of the scissile bond of the substrate and a reporter molecule, such as a fluorescence marker, to the opposite side of the scissile bond, incubating the modified substrate and the enzyme in multiple well plates, e.g. 96-well plates, optionally in the presence if a test inhibitor or activator compound transferring the incubation solutions to a second multiple well plate having upper and lower chambers separated by a porous membrane the upper chamber of which contains resin beads capable of binding with the resin-binding compound, filtering and washing the wells of the second plate and reading the emission from the plates. The invention also provides protease substrates for HIV-1 protease, vertebrate stromelysin and derivatives thereof which are useful in the assay method.