Patents by Inventor Thomas S. Wehrman
Thomas S. Wehrman has filed for patents to protect the following inventions. This listing includes patent applications that are pending as well as patents that have already been granted by the United States Patent and Trademark Office (USPTO).
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Patent number: 9388449Abstract: Methods and materials are disclosed for use in an enzyme fragment complementation assay using complementary fragments of ?-galactosidase to study the trafficking of proteins in a cell. Compounds that bind to a target peptide have been found to affect protein folding and therefore trafficking. ?-Galactosidase fragments, an enzyme donor (ED) and an enzyme acceptor (EA), are fused to a target peptide and to an intracellular compartment protein, wherein the compartment is involved in intracellular trafficking. Contacting the cell with a compound that binds to the target peptide results in enhanced movement of the protein through the cellular trafficking pathway comprised of the endoplasmic reticulum, Golgi apparatus, the plasma membrane, endosomes, etc. Using this approach, compounds that bind to a target peptide and alter its ability to traffic through the normal cellular pathway can be readily detected.Type: GrantFiled: October 21, 2013Date of Patent: July 12, 2016Assignee: DISCOVERX CORPORATIONInventors: Thomas S. Wehrman, Daniel Bassoni, William Raab
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Patent number: 8945853Abstract: Methods and compositions are provided to measure the binding of a test compound to a target peptide by measuring the effect of the compound on the abundance of the target peptide inside a cell. The target peptide may bind the test compound at an active site or an allosteric site, and it has been found that such binding may stabilize the target peptide against cellular degradation. The target peptide will preferably comprise a destabilizing mutation which shortens the half life of the target peptide within the cell, typically a mammalian cell. Test compounds, including small molecules, have been found to stabilize target peptides. Also provided are systems and kits for use in practicing the methods.Type: GrantFiled: February 5, 2013Date of Patent: February 3, 2015Assignee: DiscoveRx CorporationInventors: William Raab, Thomas S. Wehrman
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Publication number: 20140370522Abstract: Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between sub-cellular compartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing ?-galactosidase fragments are provided. These ?-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.Type: ApplicationFiled: March 24, 2014Publication date: December 18, 2014Applicant: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Helen M. Blau, Thomas S. Wehrman
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Patent number: 8865421Abstract: Methods and genetic constructs are provided for detecting the binding of nuclear hormone receptors to a coactivator/corepressor. The methods employ enzyme fragment complementation using fragments of ?-galactosidase as the detection system. Cells are transformed to express the large fragment of ?-galactosidase fused to a member of the complex with NHR for initiation of transcription and have it localized in the nucleus and to express the small fragment of ?-galactosidase fused to the nuclear hormone receptor for binding to the member upon stimulation with a ligand.Type: GrantFiled: June 23, 2009Date of Patent: October 21, 2014Assignee: DiscoveRx CorporationInventors: Thomas S. Wehrman, Chin Yee Loh, Mahesh Mathrubutham, Keith R. Olson
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Patent number: 8679832Abstract: Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between sub-cellular compartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing ?-galactosidase fragments are provided. These ?-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.Type: GrantFiled: September 20, 2011Date of Patent: March 25, 2014Assignee: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Helen M. Blau, Thomas S. Wehrman
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Publication number: 20140045194Abstract: Methods and materials are disclosed for use in an enzyme fragment complementation assay using complementary fragments of ?-galactosidase to study the trafficking of proteins in a cell. Compounds that bind to a target peptide have been found to affect protein folding and therefore trafficking. ?-Galactosidase fragments, an enzyme donor (ED) and an enzyme acceptor (EA), are fused to a target peptide and to an intracellular compartment protein, wherein the compartment is involved in intracellular trafficking. Contacting the cell with a compound that binds to the target peptide results in enhanced movement of the protein through the cellular trafficking pathway comprised of the endoplasmic reticulum, Golgi apparatus, the plasma membrane, endosomes, etc. Using this approach, compounds that bind to a target peptide and alter its ability to traffic through the normal cellular pathway can be readily detected.Type: ApplicationFiled: October 21, 2013Publication date: February 13, 2014Applicant: DiscoveRx CorporationInventors: Thomas S. Wehrman, Daniel Bassoni, William Raab
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Patent number: 8586294Abstract: Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between subcellular copartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing ?-galactosidase fragments are provided. These ?-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.Type: GrantFiled: May 18, 2005Date of Patent: November 19, 2013Assignee: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Helen M. Blau, Thomas S. Wehrman
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Patent number: 8569057Abstract: Methods and materials are disclosed for use in an enzyme fragment complementation assay using complementary fragments of ?-galactosidase to study the trafficking of proteins in a cell. Compounds that bind to a target peptide have been found to affect protein folding and therefore trafficking. ?-Galactosidase fragments, an enzyme donor (ED) and an enzyme acceptor (EA), are fused to a target peptide and to an intracellular compartment protein, wherein the compartment is involved in intracellular trafficking. Contacting the cell with a compound that binds to the target peptide results in enhanced movement of the protein through the cellular trafficking pathway comprised of the endoplasmic reticulum, Golgi apparatus, the plasma membrane, endosomes, etc. Using this approach, compounds that bind to a target peptide and alter its ability to traffic through the normal cellular pathway can be readily detected.Type: GrantFiled: June 22, 2012Date of Patent: October 29, 2013Assignee: DiscoveRx CorporationInventors: Thomas S. Wehrman, Daniel Bassoni, William Raab
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Publication number: 20120329075Abstract: Methods and materials are disclosed for use in an enzyme fragment complementation assay using complementary fragments of ?-galactosidase to study the trafficking of proteins in a cell. Compounds that bind to a target peptide have been found to affect protein folding and therefore trafficking. ?-Galactosidase fragments, an enzyme donor (ED) and an enzyme acceptor (EA), are fused to a target peptide and to an intracellular compartment protein, wherein the compartment is involved in intracellular trafficking. Contacting the cell with a compound that binds to the target peptide results in enhanced movement of the protein through the cellular trafficking pathway comprised of the endoplasmic reticulum, Golgi apparatus, the plasma membrane, endosomes, etc. Using this approach, compounds that bind to a target peptide and alter its ability to traffic through the normal cellular pathway can be readily detected.Type: ApplicationFiled: June 22, 2012Publication date: December 27, 2012Applicant: DISCOVERX CORPORATIONInventors: Thomas S. Wehrman, Daniel Bassoni, William Raab
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Patent number: 8211655Abstract: A method for determining ligand activation of receptors using cells expressing genetic constructs of a fusion protein of at least a binding domain of an auxiliary protein and a fragment of ?-galactosidase, a fusion protein of an endosome-associated protein and a complementary fragment of ?-galactosidase, and a wild-type receptor. The receptors are characterized by binding to the auxiliary protein-binding domain upon activation by an agonist and then endocytosing associated with an endosome to which the endosome-associated protein binds. Cells are incubated with a candidate ligand followed by lysis with a lysing medium comprising a substrate for the ?-galactosidase. The enzyme product is then detected as a measure of the activation of the receptor.Type: GrantFiled: February 11, 2010Date of Patent: July 3, 2012Assignee: Discoverx CorporationInventors: Thomas S. Wehrman, William Raab, Chin Yee Loh
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Patent number: 8148110Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, using at least two inactive, weakly-complementing ?-lactamase fragments are provided. The invention allows detection of such interactions in eukaryotic and mammalian cells, in situ or in vitro. Detection of molecular interactions in mammalian cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. Methods provided utilize novel compositions comprising fusion proteins between molecules of interest and inactive, weakly-complementing ?-lactamase fragments. Association of the molecules of interest brings the corresponding complementary ?-lactamase fragments into close enough proximity for complementation to occur and ?-lactamase activity to be observed. The invention is useful in the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.Type: GrantFiled: December 26, 2002Date of Patent: April 3, 2012Assignees: The Board of Trustees of the Leland Stanford Junior University, KaloBios, Inc.Inventors: Helen M. Blau, Robert F. Balint, Thomas S. Wehrman, Jeng-Horng Her
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Publication number: 20120077204Abstract: Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between sub-cellular compartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing ?-galactosidase fragments are provided. These ?-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.Type: ApplicationFiled: September 20, 2011Publication date: March 29, 2012Applicant: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Helen M. Blau, Thomas S. Wehrman
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Publication number: 20100203555Abstract: A method for determining ligand activation of receptors using cells expressing genetic constructs of a fusion protein of at least a binding domain of an auxiliary protein and a fragment of ?-galactosidase, a fusion protein of an endosome-associated protein and a complementary fragment of ?-galactosidase, and a wild-type receptor. The receptors are characterized by binding to the auxiliary protein-binding domain upon activation by an agonist and then endocytosing associated with an endosome to which the endosome-associated protein binds. Cells are incubated with a candidate ligand followed by lysis with a lysing medium comprising a substrate for the ?-galactosidase. The enzyme product is then detected as a measure of the activation of the receptor.Type: ApplicationFiled: February 11, 2010Publication date: August 12, 2010Applicant: DISCOVERX CORPORATIONInventors: Thomas S. Wehrman, William Raab, Chin Yee Loh
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Publication number: 20100151496Abstract: Methods and genetic constructs are provided for detecting the binding of nuclear hormone receptors to a coactivator/corepressor. The methods employ enzyme fragment complementation using fragments of ?-galactosidase as the detection system. Cells are transformed to express the large fragment of ?-galactosidase fused to a member of the complex with NHR for initiation of transcription and have it localized in the nucleus and to express the small fragment of ?-galactosidase fused to the nuclear hormone receptor for binding to the member upon stimulation with a ligand.Type: ApplicationFiled: June 23, 2009Publication date: June 17, 2010Applicant: DiscoveRx CorporationInventors: Thomas S. Wehrman, Chin Yee Loh, Mahesh Mathrubutham, Keith R. Olson
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Publication number: 20100120063Abstract: Sensitive assays for candidate compounds affecting GPCR activity are provided using a cell containing fusion proteins comprising a first fusion protein comprising (a) a target GPCR fused to a small fragment of ?-galactosidase through a linker comprising a phosphorylation site or (b) a GPCR or a protein of interest, where the GPCR and protein of interest form a complex and one of them is fused to the small fragment of ?-galactosidase; and a second fusion protein comprising arrestin fused to a large fragment of ?-galactosidase. In (a), the affinity of the small and large fragments is optimized based on the background to signal ratio and the absolute signal observed. The assay is performed using a ?-galactosidase substrate that provides a detectable optical signal.Type: ApplicationFiled: October 10, 2009Publication date: May 13, 2010Applicant: DISCOVERX CORPORATIONInventors: Daniel Bassoni, Keith R. Olson, Thomas S. Wehrman
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Publication number: 20030175836Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, using at least two inactive, weakly-complementing &bgr;-lactamase fragments are provided. The invention allows detection of such interactions in eukaryotic and mammalian cells, in situ or in vitro. Detection of molecular interactions in mammalian cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. Methods provided utilize novel compositions comprising fusion proteins between molecules of interest and inactive, weakly-complementing &bgr;-lactamase fragments. Association of the molecules of interest brings the corresponding complementary &bgr;-lactamase fragments into close enough proximity for complementation to occur and &bgr;-lactamase activity to be observed. The invention is useful in the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.Type: ApplicationFiled: December 26, 2002Publication date: September 18, 2003Inventors: Helen M. Blau, Robert F. Balint, Thomas S. Wehrman, Jeng-Horng Her