Abstract: 196. Disclosed are methods of incubating cells on biosensors, and methods using the disclosed incubation techniques to identify PDE4 modulators.

Abstract: The present invention includes several methods for modifying the current processes of manufacturing optical sensing microplates that use continuous waveguide films to reduce/eliminate crosstalk between the biosensors that are incorporated within wells. The methods include (1) physically deteriorating/removing the waveguide film between individual biosensors; (2) chemically depositing highly absorbing materials within the waveguide film between individual biosensors; (3) patterning disordered (scattering) regions between the diffraction gratings that define individual biosensors; (4) using a specific mask and depositing individual patches of waveguide film, where each patch defines at least one biosensor. Each of these methods and several other methods described herein prevent the propagation of light between individual sensing regions, thereby eliminating optical crosstalk between the biosensors. The present invention also includes the resulting microplate.

Abstract: A substrate plate or device adapted for use with biological or chemical assays is disclosed. The device may take the form of a multi-well plate having a three-dimensional, porous layer as part of a support surface within each well for immobilizing probe species. The porous layer is characterized as having a plurality of interconnected voids defined by a matrix of contiguous solid material. A method and its variants are also described.

Abstract: A substrate plate or device adapted for use with biological or chemical assays is disclosed. The device may take the form of a multi-well plate having a three-dimensional, porous layer as part of a support surface within each well for immobilizing probe species. The porous layer is characterized as having a plurality of interconnected voids defined by a matrix of contiguous solid material. A method and its variants are also described.

Abstract: Disclosed is a system and method for measuring aspects of pathogen intrusion on a live-cell as defined herein. The system and method also provide a method to measure prophylaxis or remedial aspects of a therapeutic candidates in a live-cell or a live-cell model from pathogen intrusion.

Abstract: Methods for performing surface-mediated protein delivery into living cells, and fabricating protein-transfected cell cluster arrays are provided. The method comprises providing a protein-containing mixture; depositing said protein-containing mixture onto a surface at defined locations; affixing the protein-containing mixture to the surface as microspots; and plating cells onto the surface in sufficient density and under conditions for the proteins to be delivered into the cells. The protein-containing mixture comprises any suitable amino acid sequence, including peptides, proteins, protein-domains, antibodies, or protein-nucleic acid conjugates, etc., with a carrier reagent. Protein-transfected cell arrays may be used for rapid and direct, screening of protein or enzymatic functions or any given intracellular protein interaction in the natural environment of a living cell, as well as for high-throughput screening of other biological and chemical analytes, which affect the functions of these proteins.

Abstract: Buffered assay solutions for performing 1) binding or 2) functional assays on GPCR arrays, along with methods for their use are described. The buffered assay solution has an underlying composition having: a buffer reagent with a pH in the range of about 6.5 to about 7.9; an inorganic salt of either a monovalent or divalent species, at a concentration from about 1 mM to about 500 mM; and optionally a combination of: c) a blocker reagent at a concentration of about 0.01 wt. % to about 2 wt. % of the composition, or d) protease-inhibitor at a concentration of about 0.001 mM to about 100 mM. In an embodiment for functional assay uses, the composition is modified to also include a GTP-analogue, a guanosine 5?-diphosphate (GDP) salt, and/or an anti-oxidant reagent.

Abstract: The present invention overcomes the problems and disadvantages associated with prior art arrays by providing an array comprising a plurality of biological membrane microspots associated with a surface of a substrate that can be produced, used and stored, not in an aqueous environment, but in an environment exposed to air under ambient or controlled humidities. Preferably, the biological membrane microspots comprise a membrane bound protein. Most preferably, the membrane bound protein is a G-protein coupled receptor, an ion channel, a receptor serine/threonine kinase or a receptor tyrosine kinase.

Abstract: A method and apparatus for implementing the method is provided. The method involves performing an indirect competitive binding assay on a microarray to identify biological or chemical targets and screen for compounds of interest. The microarray comprises a common ligand located among membrane-, lipid- or protein-associated active binding sites. The method takes advantage of known or well-characterized binding kinetics, and steric interference between biological or chemicals targets of interest and universal readout units for different binding sites within the limited confines of a microspot. The biological targets, chemicals or organisms can specifically bind to target-binding sites, while the universal readout unit binds to the ligands in the microspot.

Abstract: Buffered assay solutions for performing 1) binding or 2) functional assays on GPCR arrays, along with methods for their use are described. The buffered assay solution has an underlying composition having: a buffer reagent with a pH in the range of about 6.5 to about 7.9; an inorganic salt of either a monovalent or divalent species, at a concentration from about 1 mM to about 500 mM; and optionally a combination of: c) a blocker reagent at a concentration of about 0.01 wt. % to about 2 wt. % of the composition, or d) protease-inhibitor at a concentration of about 0.001 mM to about 100 mM. In an embodiment for functional assay uses, the composition is modified to also include a GTP-analogue, a guanosine 5?-diphosphate (GDP) salt, and/or an anti-oxidant reagent.

Abstract: The disclosure provides methods for characterizing the toxicity of a candidate molecule to liver cells as defined herein; methods of culturing metabolically active liver cells on a biosensor as defined herein; and biosensor liver culture systems as defined herein.

Abstract: Drug discovery is a complex undertaking facing many challenges, not the least of which is a high attrition rate as many promising candidates prove ineffective or toxic in the clinic owing to a poor understanding of the diseases, and thus the biological systems, they target. Therefore, it is broadly agreed that to increase the productivity of drug discovery one needs a far deeper understanding of the molecular mechanisms of diseases, taking into account the full biological context of the drug target and moving beyond individual genes and proteins. The present methods rely on the use of label-free cellular assays, particularly the DMR index, to systematically display the mode of actions, the toxicity, and the target(s) and pathway(s) of any molecules.

Abstract: Drug discovery is a complex undertaking facing many challenges, not the least of which is a high attrition rate as many promising candidates prove ineffective or toxic in the clinic owing to a poor understanding of the diseases, and thus the biological systems, they target. Therefore, it is broadly agreed that to increase the productivity of drug discovery one needs a far deeper understanding of the molecular mechanisms of diseases, taking into account the full biological context of the drug target and moving beyond individual genes and proteins. The present methods rely on the use of label-free cellular assays, particularly the DMR index, to systematically display the mode of actions, the toxicity, and the target(s) and pathway(s) of any molecules.

Abstract: Aspects relate to the field of label-free biosensors, including optical biosensors and electric biosensors, for studying cellular behaviour particularly early events in cell migration, under a concentration gradient of stimulus. Various embodiments include devices and methods that enable the generation of a concentration gradient of stimulus for cells contacting with the surface of a biosensor including a label free biosensor. Some aspects also disclose methods to detect cellular responses upon such concentration gradient of a stimulus, as well as methods to detect the potency and efficacy of a stimulus using a single biosensor.

Abstract: An article for culturing cells includes a substrate on which cells can be cultured. The substrate has a base surface. An array of projections extends from the base surface. The projections have a height of about 1 micrometer to about 100 micrometers, and have a gap distance along the major surface from center to center between neighboring projections of about 10 micrometers to 80 micrometers. A plurality of arrays of projections may extend from the surface with gaps in the base surface between the arrays. Hepatocytes cultures on such microprojection array substrates maintained in vivo like morphology and membrane polarity. Hepatocytes co-cultured with helper cells on such substrates tended to grow in the area of the arrays, while the helper cells tended to grow in the areas between the arrays.

Abstract: The disclosure provides a device for measuring cellular responses under controlled environments. The disclosure also provides an array of devices for measuring cellular responses under controlled environments. The disclosure also provides methods to study cell signaling using the devices. By using microfluidics, the disclosure enables study of cell signaling under well-defined environment conditions. Such capability can be used for differentiating long-acting ligands from short-acting ligands, for determining the kinetics of receptor resensitization and functional recovery of receptor signaling, and for differentiating the sensitivity of a cell type to laminar flow-induced stress force. Through combination with conventional static label-free cell assays, the full spectrum of a drug compound can be studied in details, thus creating a complete representation of its pharmacology acting on living cells.

Abstract: A method for hit compound identification in a high throughput, label-free biosensor cellular assay, one method including, for example: generating at least one pathway-specific kinetic profile for a target receptor; generating a relational table that integrates each kinetic profile; determining a delta response with the relational table for candidate compounds screened against the target receptor; and identifying one or more hit compounds based upon the separation of the delta response from a threshold.

Abstract: Systems, methods and kits that utilize uniquely coded microparticles for performing protein assays are provided. The uniquely coded microparticles are used as a substrate for reverse protein delivery into cells. The microparticles and methods offer the possibility of studying the biological functions of either a single protein of interest in multiple cell types per assay or multiple proteins in a single cell type.

Abstract: Buffered assay solutions for performing 1) binding or 2) functional assays on GPCR arrays, along with methods for their use are described. The buffered assay solution has an underlying composition having: a buffer reagent with a pH in the range of about 6.5 to about 7.9; an inorganic salt of either a monovalent or divalent species, at a concentration from about 1 mM to about 500 mM; and optionally a combination of: c) a blocker reagent at a concentration of about 0.01 wt. % to about 2 wt. % of the composition, or d) protease-inhibitor at a concentration of about 0.001 mM to about 100 mM. In an embodiment for functional assay uses, the composition is modified to also include a GTP-analogue, a guanosine 5?-diphosphate (GDP) salt, and/or an anti-oxidant reagent.

Abstract: The present invention overcomes the problems and disadvantages associated with prior art arrays by providing an array comprising a plurality of biological membrane microspots associated with a surface of a substrate that can be produced, used and stored, not in an aqueous environment, but in an environment exposed to air under ambient or controlled humidities. Preferably, the biological membrane microspots comprise a membrane bound protein. Most preferably, the membrane bound protein is a G-protein coupled receptor, an ion channel, a receptor serine/threonine kinase or a receptor tyrosine kinase.