C - Chemistry – Metallurgy – 12 – Q
Patent
C - Chemistry, Metallurgy
12
Q
C12Q 1/34 (2006.01) C07K 7/06 (2006.01) C07K 7/08 (2006.01) C07K 7/22 (2006.01) C07K 14/16 (2006.01) C07K 14/78 (2006.01) C12Q 1/37 (2006.01) G01N 33/573 (2006.01)
Patent
CA 2070719
X-7650 Abstract 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 is 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.
Heath William F. Jr.
Lai Mei-Huei T.
Manetta Joseph V.
Sportsman John R.
Yan Sau-Chi B.
Eli Lilly And Company
Gowling Lafleur Henderson Llp
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