Membrane receptor reagent and assay:
Patent 6,790,632
& 7,288,368. 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.
Synthetic substrate
for high specificity enzymatic assays: Patent7,291,698(CIP in progress).
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.
Apoenzyme
reactivation electrochemical detection method and assay: Patent
7,166,208(CIP in progress) The invention discloses methods in which dry reagent enzyme
based electrochemical biosensors, which are in a relatively mature form due to
the extensive amount of development pioneered by the blood glucose monitoring
industry, may be simply adapted to perform tests for blood coagulation,
enzymatic activity, or immunochemical assays for antigens present in a fluid
sample. In particular, the utility of combining apoenzyme based dry reagent
electrochemical biosensors with apoenzyme reactivation technology is taught.
This combination creates a novel combination dry reagent test technology capable
of detecting a wide range of different analytes.
Tethered receptor-ligand
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.
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