Membrane receptor reagent and assay: Patent 6,790,632
(text). A membrane receptor reagent and assay is disclosed in which
liposomes are bound to an evanescent wave emitting surface. Membrane receptors
on the liposome's fluid lipid bilayer membrane are labeled with a fluorescent or
luminescent moiety. These membrane receptors are free to diffuse randomly
throughout the liposome surface, and thus tend to redistribute according to
externally applied forces. The evanescent wave-emitting surface additionally
contains reagents that reversibly bind to the membrane receptors, tending to
bring them closer to region of high evanescent wave intensity. Test analytes
that disrupt or promote the association between the membrane receptors and the
surface reagents act to change the average distance between the membrane
receptors and the evanescent wave emitting surface, resulting in a change in the
fluorescent or luminescent signal. This reagent and assay system functions with
physiologically important membrane receptors such as GPCR receptors, other 7-tm
receptors, drug transport proteins, cytochrome P450 membrane proteins and other
clinically important membrane components. The reagent and assay methods may be
incorporated into microarrays, capillaries, flow cells and other devices, and
used for drug discovery, ADMET, and other biomedically important assays.
reagent and assay: US application 20030108972 (text).
A tethered reagent and assay is
disclosed consisting of protein receptors tethered to ligands. The protein
receptors can be antibodies, enzymes, hormone receptors, integral membrane
proteins, and other proteins. Ligands can be antigens, enzymatic inhibitors,
hormone agonists, drugs, and other protein binding ligands. The protein
receptors and ligands will each be labeled with moieties capable of detecting
changes in the average distance between the protein receptors and the ligand,
using detection methods in which there is a sharp fall-off in signal as a
function of distance. As a result, a change in the average distance between the
two label moieties, such as that caused by protein-ligand binding and
dissociation, produces a change in a detectable signal produced by the reagent.
Tethering means may consist of flexible polymers, typically composed of a
material that is chemically distinct from either the receptor or the ligand, so
that the receptors and ligands may freely associate and dissociate via their
specific binding sites, but not totally diffuse away from each other. When bound
to solid phase surfaces, such reagents are particularly well suited for
proteomic microarrays and flow cells. Such reagents may have utility for
immunoassays, enzyme assays, ligand binding assays, sepsis assays, drug
screening assays, and drug ADMET assays.
for high specificity enzymatic assays: US application 20030113768 (text).
Novel synthetic enzyme substrates, enhanced to have improved enzymatic
specificity, are disclosed. These synthetic enzyme substrates consist of a
substrate peptide that has had its specificity further improved by additional
synthetic moieties, selected by combinatorial chemistry techniques, that act to
sterically block non-target enzymes. These "steric restrictor" moieties may be
labeled to produce a detectable signal upon enzymatic reaction. These novel
substrates are particularly useful for improved enzyme substrate microarrays.
Specific applications for improved protease substrate microarrays are discussed.
A variety of applications for these improved protease substrate microarrays are
also disclosed, including proteomics research, protease discovery, protease
binding site characterization, diagnosis of the protease composition of
biological samples, monitoring the angiogenic status of a tumor, monitoring the
status of arthritis and other inflammatory diseases, and the discovery and
optimization of novel drugs that modify or inhibit protease activity.
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