Identifying A Library Member By Means Of A Tag, Label, Or Other Readable Or Detectable Entity Associated With The Library Member (e.g., Decoding Process, Etc.) Patents (Class 506/4)
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Patent number: 8211428Abstract: Methods for identifying modified proteases with modified substrate specificity or other properties are provided. The methods screen candidate and modified proteases by contacting them with a substrate, such as a serpin, an alpha macroglobulins or a p35 family protein or modified serpins and modified p35 family members or modified alpha macroglobulins, that, upon cleavage of the substrate, traps the protease by forming a stable complex. Also provided are modified proteases.Type: GrantFiled: July 5, 2007Date of Patent: July 3, 2012Assignees: Torrey Pines Institute for Molecular Studies, Catalyst Biosciences, Inc.Inventor: Edwin L. Madison
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Publication number: 20120142540Abstract: Methods and compositions for determining the nucleic acid sequence of polynucleotides that are at least 1500 nucleotides in length are provided.Type: ApplicationFiled: November 4, 2011Publication date: June 7, 2012Applicant: President and Fellows of Harvard CollegeInventors: Chao-ting Wu, George M. Church
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Publication number: 20120071329Abstract: The present invention provides a method for identifying a compound of interest by screening libraries of molecules which include an encoding oligonucleotide tag.Type: ApplicationFiled: March 18, 2011Publication date: March 22, 2012Applicant: GlaxoSmithKline LLCInventors: Barry Morgan, Stephen Hale, Christopher C. Arico-Muendel, Matthew Clark, Richard Wagner, David I. Israel, Malcolm L. Gefter, Dennis Benjamin, Nils Jakob Vest Hansen, Malcolm J. Kavarana, Steffen Phillip Creaser, George J. Franklin, Paolo A. Centrella, Raksha A. Acharya
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Publication number: 20120071330Abstract: A method for the rapid identification of a target nucleic acid sequence is provided, as well as corresponding devices, products and kits. Such methods are useful for the rapid detection, identification and/or quantification of target nucleic acid sequences associated with, for example, a pathogen.Type: ApplicationFiled: June 30, 2011Publication date: March 22, 2012Applicant: Stratos Genomics Inc.Inventors: Mark Stamatios Kokoris, Robert N. McRuer
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Publication number: 20120071331Abstract: Aspects of the present invention include methods and compositions for determining the number of individual polynucleotide molecules originating from the same genomic region of the same original sample that have been sequenced in a particular sequence analysis configuration or process. In these aspects of the invention, a degenerate base region (DBR) is attached to the starting polynucleotide molecules that are subsequently sequenced (e.g., after certain process steps are performed, e.g., amplification and/or enrichment). The number of different DBR sequences present in a sequencing run can be used to determine/estimate the number of different starting polynucleotides that have been sequenced. DBRs can be used to enhance numerous different nucleic acid sequence analysis applications, including allowing higher confidence allele call determinations in genotyping applications.Type: ApplicationFiled: September 20, 2011Publication date: March 22, 2012Inventors: James Casbon, Sydney Brenner, Robert Osborne, Conrad Lichtenstein
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Publication number: 20120046178Abstract: Provided herein are products and processes for detecting the presence or absence of multiple target nucleic acids. Certain methods include amplifying the target nucleic acids, or portion thereof; extending oligonucleotides that specifically hybridize to the amplicons, where the oligonucleotides include distinguishable labels and a capture agent; capturing the extended oligonucleotides to a solid phase via the capture agent; releasing and detecting the distinguishable label, and thereby determining the presence or absence of each target nucleic acid by the presence or absence of the distinguishable label.Type: ApplicationFiled: October 27, 2009Publication date: February 23, 2012Applicant: SEQUENOM, INC.Inventors: Dirk Johannes Van Den Boom, Christiane Honisch, Andrew Timms, Smita Chitnis
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Patent number: 8105982Abstract: The present method is an improved in vitro selection protocol that relies on magnetic separations for DNA aptamer production that is relatively easy and scalable without the need for expensive robotics. The ability of aptamers selected by this method to recognize and bind their target protein with high affinity and specificity, and detail their uses in a number of assays is also described. Specific TTF1 and His6 aptamers were selected using the method described, and shown to be useful for enzyme-linked assays, Western blots, and affinity purification.Type: GrantFiled: February 6, 2008Date of Patent: January 31, 2012Assignee: The Regents of the University of CaliforniaInventors: Sharon A. Doyle, Michael B. Murphy
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Method for performing a multiplex immunoassay using label disassociation and an integrated substrate
Patent number: 8097421Abstract: The application relates to a method, system and device for performing biological assays. The method, system and device allow more accurate and specific detections of biomolecules in multiplex assays, such as immunoassays and DNA microarray assays. More specifically, the embodiments of the invention allow the detection of labels after their detachment or disassociation from a binding situation wherein interference from other labels or the background may reduce the accuracy of specificity of the detection. The embodiments of the invention further allow detection of individual labels.Type: GrantFiled: December 29, 2005Date of Patent: January 17, 2012Assignee: Intel CorporationInventor: Tae-Woong Koo -
Publication number: 20110301044Abstract: A system and method for imaging biological samples on multiple surfaces of a support structure are disclosed. The support structure may be a flow cell through which a reagent fluid is allowed to flow and interact with the biological samples. Excitation radiation from at least one radiation source may be used to excite the biological samples on multiple surfaces. In this manner, fluorescent emission radiation may be generated from the biological samples and subsequently captured and detected by detection optics and at least one detector. The detected fluorescent emission radiation may then be used to generate image data. This imaging of multiple surfaces may be accomplished either sequentially or simultaneously. In addition, the techniques of the present invention may be used with any type of imaging system. For instance, both epifluorescent and total internal reflection methods may benefit from the techniques of the present invention.Type: ApplicationFiled: August 12, 2011Publication date: December 8, 2011Applicant: ILLUMINA, INC.Inventors: Wenyi Feng, Jason Bryant, Dale Buermann
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Publication number: 20110301043Abstract: The present invention provides a method of predicting the risk of a patient for developing cutaneous adverse drug reaction to non-nucleoside reverse transcriptase inhibitors such as nevirapine by using HLA-B*3505 allele and/or polymorphisms in the CCHCR1 gene.Type: ApplicationFiled: December 11, 2009Publication date: December 8, 2011Applicants: MAHIDOL UNIVERSITY, RIKEN, DEPARTMENT OF MEDICAL SCIENCESInventors: Somnuek Sungkanuparph, Sasisopin Kiertiburanakul, Thanyachai Sura, Wasun Chantratita, Soranun Chantarangsu, Angkana Charoenyingwattana, Surakameth Mahasirimongkol, Michiaki Kubo, Taisei Mushiroda, Yusuke Nakamura
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Publication number: 20110301042Abstract: The invention generally relates to methods for sequencing a plurality of nucleic acids from different samples. In certain embodiments, methods of the invention provide contacting a nucleic acid duplex including a primer nucleic acid hybridized to a template nucleic acid with a polymerase enzyme in the presence of a first detectably labeled nucleotide under conditions that permit the polymerase to add nucleotides to the primer in a template-dependent manner, in which a unique oligonucleotide sequence is attached to the template nucleic acid so that the template nucleic acid may be differentiated from other template nucleic acid molecules, detecting a signal from the incorporated labeled nucleotide, and sequentially repeating the contacting and detecting steps at least once, wherein sequential detection of incorporated labeled nucleotide determines the sequence of the nucleic acid.Type: ApplicationFiled: November 11, 2009Publication date: December 8, 2011Applicant: HELICOS BIOSCIENCES CORPORATIONInventors: Kathleen Steinmann, Elaine Joseph, Marie Causey, John F. Thompson
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Patent number: 8017559Abstract: The present invention provides novel polypeptides having the scaffold structure of a C-type lectin-like domain (CTLD) and a randomized loop region for specifically binding a variety of target compounds and also provides nucleic acids encoding the polypeptides. The present invention further provides combinatorial CTLD libraries, methods for constructing the libraries, and methods for screening the libraries to identify and isolate the novel CTLD polypeptides. Specifically, the invention provides libraries of nucleic acids encoding polypeptides having a scaffold CTLD with a randomized loop region, as well as nucleic acid sequences, vectors, and methods for preparing and expressing the libraries. Exemplary nucleic acids useful in the combinatorial libraries are derived from tetranectin and other proteins having a CTLD.Type: GrantFiled: December 4, 2006Date of Patent: September 13, 2011Assignee: Anaphore, Inc.Inventors: Michael Etzerodt, Thor Las Holtet, Niels Jonas Heilskov Graversen, Hans Christian Thøgersen
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Patent number: 7998904Abstract: Nature evolves biological molecules such as proteins through iterated rounds of diversification, selection, and amplification. The present invention provides methods, compositions, and systems for synthesizing, selecting, amplifying, and evolving non-natural molecules based on nucleic acid templates. The sequence of a nucleic acid template is used to direct the synthesis of non-natural molecules such as unnatural polymers and small molecules. Using this method combinatorial libraries of these molecules can be prepared and screened. Upon selection of a molecule, its encoding nucleic acid template may be amplified and/or evolved to yield the same molecule or related molecules for re-screening. The inventive methods and compositions of the present invention allow for the amplification and evolution of non-natural molecules in a manner analogous to the amplification of natural biopolymer such as polynucleotides and protein.Type: GrantFiled: May 31, 2005Date of Patent: August 16, 2011Assignee: President and Fellows of Harvard CollegeInventors: David R. Liu, Zev J. Gartner, Matthew W. Kanan
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Publication number: 20110166027Abstract: An interaction screening method for identifying binding moieties encapsulates candidate binding moieties in droplets with a first known moiety and a second known moiety. The candidate binding moieties are different in the different droplets. The method further comprises determining for one or more of the droplets whether the candidate binding moiety is bound to the first known moiety and/or the second known moiety. Optionally, the method further comprises segregating at least one droplet in which the candidate binding moiety is bound to the first known moiety or to the first and second known moiety.Type: ApplicationFiled: August 21, 2008Publication date: July 7, 2011Applicant: AFFOMIX CORPORATIONInventor: Michael P. Weiner
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Patent number: 7964534Abstract: The invention provides a method for identifying a ligand that binds to a macromolecular target. The methods involve (a) attaching an antenna moiety to a first ligand, wherein the ligand binds specifically to a macromolecular target; (b) providing a sample comprising the macromolecular target, the first ligand and a candidate second ligand under conditions wherein the first ligand and the macromolecular target form a bound complex; (c) detecting a subset of magnetization transfer signals between the antenna moiety of the first ligand and the second candidate ligand, wherein the signals are obtained from an isotope edited NOESY spectrum of the sample; thereby determining that the two ligands are proximal in a bound complex, and identifying a second ligand that binds to the macromolecular target.Type: GrantFiled: March 12, 2004Date of Patent: June 21, 2011Assignee: Triad Liquidating Company, LLCInventors: Mark Kelly, Hugo Villar, Jianqiang Wang, Min S. Lee, Yong Qin, Daniel S. Sem
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Publication number: 20110136676Abstract: The invention is related to nucleic acid arrays and methods of using nucleic acid arrays.Type: ApplicationFiled: October 25, 2010Publication date: June 9, 2011Inventor: Eric C. Greene
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Publication number: 20110118125Abstract: The present invention provides systems for assessing neonatal development and/or conditions by analyzing neonatal saliva RNA. Methods of identifying genes involved in neonatal development and/or conditions affecting neonates, are provided. Methods of determining a diagnosis of a neonate comprising detection of one or more differentially expressed genes are also provided.Type: ApplicationFiled: May 1, 2009Publication date: May 19, 2011Inventors: Jill Maron, Diana Bianchi, Kirby Johnson, Donna Slonim
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Publication number: 20110118148Abstract: The invention provides a method for identifying a ligand that binds to a macromolecular target. The methods involve (a) attaching an antenna moiety to a first ligand, wherein the ligand binds specifically to a macromolecular target; (b) providing a sample comprising the macromolecular target, the first ligand and a candidate second ligand under conditions wherein the first ligand and the macromolecular target form a bound complex; (c) detecting a subset of magnetization transfer signals between the antenna moiety of the first ligand and the second candidate ligand, wherein the signals are obtained from an isotope edited NOESY spectrum of the sample; thereby determining that the two ligands are proximal in a bound complex, and identifying a second ligand that binds to the macromolecular target.Type: ApplicationFiled: March 12, 2004Publication date: May 19, 2011Inventors: Mark Kelly, Hugo Villar, Jianqiang Wang, Min S. Lee, Yong Qin, Daniel S. Sem
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Patent number: 7935659Abstract: Combinations, called matrices with memories, of matrix materials with remotely addressable or remotely programmable recording devices that contain at least one data storage unit are provided. The matrix materials are those that are used in as supports in solid phase chemical and biochemical syntheses, immunoassays and hybridization reactions. The matrix materials may additionally include fluophors or other luminescent moieties to produce luminescing matrices with memories. The data storage units are non-volatile antifuse memories or volatile memories, such as EEPROMS, DRAMS or flash memory. By virtue of this combination, molecules and biological particles, such as phage and viral particles and cells, that are in proximity or in physical contact with the matrix combination can be labeled by programming the memory with identifying information and can be identified by retrieving the stored information. Combinations of matrix materials, memories, and linked molecules and biological materials are also provided.Type: GrantFiled: December 14, 2004Date of Patent: May 3, 2011Assignee: Nexus Biosystems, Inc.Inventors: Michael P. Nova, Andrew E. Senyei, Zahra Parandoosh, Gary S. David, Xiao-Yi Xiao
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Publication number: 20110082046Abstract: The present invention relates to a method for the manipulation for an identification purpose of a microcarrier comprising the steps of: (a) an identification purpose step of the microcarrier; and (b) a positioning and orientation step prior to or during the identification purpose step, wherein the identification purpose step is a detection step for the detection of an encoded microcarrier having an anisotropy in its shape. The invention further relates to an apparatus for the manipulation for identification purposes of a microcarrier comprising means for identification purposes such as a microscoper and means for the positioning and orientation of the microcarriers.Type: ApplicationFiled: September 22, 2010Publication date: April 7, 2011Applicant: Universiteit GentInventors: Marc Jan René LEBLANS, Emmanuel Marie Paul Ernest Gustin, Christiaan Hubert Simon Roelant, Stefaan Cornelis De Smedt, Joseph Demeester, Kevin Braeckmans
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Patent number: 7915201Abstract: The invention relates to a method for synthesising a bifunctional complex comprising an encoded molecule and an identifier polynucleotide identifying the chemical entities having participated in the synthesis of the encoded molecule, said method comprising the steps of i) providing a) at least one template comprising one or more codons capable of hybridising to an anti-codon, wherein said template is optionally associated with one or more chemical entities, and b) a plurality of building blocks each comprising an anti-codon associated with one or more chemical entities, and ii) hybridising the anti-codon of one or more of the provided building blocks to the template, iii) covalently linking said anti-codons and/or linking the at least one template with the anti-codon of at least one building block, thereby generating an identifier polynucleotide capable of identifying chemical entities having participated in the synthesis of the encoded molecule, iv) separating the template from one or more of the anti-codonsType: GrantFiled: March 22, 2004Date of Patent: March 29, 2011Assignee: Nuevolution A/SInventors: Thomas Franch, Soeren Nyboe Jacobsen, Torben Ravn Rasmussen, Soeren Neve, Henrik Pedersen, Alex Haahr Gouliaev
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Patent number: 7902122Abstract: A method for detecting a target biological material using DNA barcodes is provided. The method is for detecting a target biological material (e.g., DNA) by using DNA barcodes by which a trace amount of target biological material can be detected in a rapid and economic manner without performing polymerase chain reaction (PCR). The method is characterized by the use of magnetic particles and polymer particles coated with DNA barcodes to sense a trace amount of a biological material (e.g., DNA).Type: GrantFiled: June 6, 2007Date of Patent: March 8, 2011Assignee: Electronics and Telecommunications Research InstituteInventors: Sang Don Jung, Myung Ae Chung, Hyo Bong Hong
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Patent number: 7902121Abstract: T cells are profiled with respect to their expression of antigen receptor. The cells are arrayed on a planar or three-dimensional substrate through binding to immobilized or partially diffused MHC-antigen complexes. The cells may further be characterized with respect to their ability to respond to external stimulus in the microenvironment. External stimuli include cell-cell interactions, response to factors, and the like.Type: GrantFiled: May 27, 2004Date of Patent: March 8, 2011Assignee: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Daniel Shin-Yu Chen, Yoav Soen, Daniel Lewis Kraft, Patrick O. Brown, Mark Davis
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Publication number: 20110034340Abstract: The present invention discloses systems and methods for minimizing or eliminating steps in immunosorbent assays for single and multiple analytes by eliminating both the need to attach target molecules to the test well and the need to remove unbound antibodies through rinsing. The immunosorbent assay (ISA) is utilized for a single analyte or target and includes the step of mixing the immunologic molecules with the sample and detection. The present invention further discloses an immunosorbent assay for multiple analytes (ISAMA) for testing a plurality of analytes or targets in a single well using a modified ISA test wherein different tags are attached to different antibody pairs. Alternate embodiments use multiple types of scavenger antigens with corresponding elimination of the need for scavenger antibodies. The present invention discloses various types of test wells for the rapid and simultaneous testing of fluids for a plurality of components and a methodology for a continuous immunosorbent assay.Type: ApplicationFiled: August 2, 2010Publication date: February 10, 2011Inventors: Derrick Charbonnet, Norman Scott Evans
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Publication number: 20100317531Abstract: Nucleosides and nucleotides are disclosed that are linked to detectable labels via a cleavable linker group.Type: ApplicationFiled: July 13, 2010Publication date: December 16, 2010Inventors: Shankar Balasubramanian, Colin Barnes, Xiaohai Liu
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Publication number: 20100298152Abstract: The present invention is a method for measuring the amount of at least one molecule in a biological sample, the method comprising a) combining the sample, or a derivative thereof, with one or more aptamers and allowing one or more molecules in the sample to bind to the aptamer(s); b) separating bound from unbound molecules; and c) quantifying the molecule(s) bound to the or each aptamer, wherein quantification of the bound molecule(s) is carried out by sequencing at least part of the or each aptamer. Uses of and products derived from the method are also contemplated.Type: ApplicationFiled: October 10, 2008Publication date: November 25, 2010Applicant: PRONOTA NVInventors: Clive Gavin Brown, Koen Kas, Sven Agnes Jan Eyckerman
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Publication number: 20100293662Abstract: A group of genes including GhCIR1 from cotton (Gossypium hirsutum), and AtCIR1 and AtCIR2 from Arabidopsis thaliana promote shoot regeneration in plants even in the absence of cytokinin. In the presence of cytokinin, the genes significantly improve transformation efficiency. The genes can be used as an enhancer as well as a selectable marker of transformation in plants. The proteins encoded by the novel genes have a homeodomain (HD) at the N-terminus and a highly divergent domain at the C-terminus. The proteins share a common structural motif.Type: ApplicationFiled: February 13, 2006Publication date: November 18, 2010Applicant: TEMASEK LIFE SCIENCES LABORATORYInventors: Lianghui Ji, Lin Cai, Nam-Hai Chua
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Publication number: 20100173786Abstract: The present invention relates to a method for absolute quantification of polypeptides.Type: ApplicationFiled: June 2, 2008Publication date: July 8, 2010Applicant: COMMISSARIAT A L'ENERGIE ATOMIQUEInventors: Virginie Brun, Alain Dupuis, Jerome Garin
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Publication number: 20100173787Abstract: The present invention describes a method for identifying one or more of a plurality of sequences differing by one or more single base changes, insertions, deletions, or translocations in a plurality of target nucleotide sequences. The method includes a ligation phase, a capture phase, and a detection phase. The ligation phase utilizes a ligation detection reaction between one oligonucleotide probe, which has a target sequence-specific portion and an addressable array-specific portion, and a second oligonucleotide probe, having a target sequence-specific portion and a detectable label. After the ligation phase, the capture phase is carried out by hybridizing the ligated oligonucleotide probes to a solid support with an array of immobilized capture oligonucleotides at least some of which are complementary to the addressable array-specific portion. Following completion of the capture phase, a detection phase is carried out to detect the labels of ligated oligonucleotide probes hybridized to the solid support.Type: ApplicationFiled: March 16, 2010Publication date: July 8, 2010Applicant: CORNELL RESEARCH FOUNDATION, INC.Inventors: Francis BARANY, George BARANY, Robert P. HAMMER, Maria KEMPE, Herman BLOK, Monib ZIRVI
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Publication number: 20100152050Abstract: An imaging system for exciting and measuring fluorescence on or in samples comprising fluorescent materials (e.g. fluorescent labels, dyes or pigments). In one embodiment, a device is used to detect fluorescent labels on nucleic acid. In a preferred embodiment, the device is configured such that fluorescent labels in a plurality of different DNA templates are simultaneously detected.Type: ApplicationFiled: February 12, 2010Publication date: June 17, 2010Inventors: Steven Gordon, Steven Auger, Alan Schinazi
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Patent number: 7718575Abstract: A method of selecting a test molecule that binds to HLA-DP4 by (i) incubating HLA-DP4 with the test molecule and a labeled peptide of formula (I) Z1X1X2X3X4X5X6X7X8X9Z2 thereby forming respective complexes, wherein Z1 and Z2, which may be identical or different, are each either zero or from 1 to 100 amino acids; X1 is an aromatic or hydrophobic amino acid, or S; X6 is an aromatic or hydrophobic amino acid, or C; X9 is an aromatic or hydrophobic amino acid, or C, D, Q, S, T, or E; and X2, X3, X4, X5, X7and X8 are each an amino acid; (ii) separating the respective complexes formed; (iii) detecting the HLA-DP4/labeled peptide complexes; and (iv) selecting the test molecule that exhibits a binding activity of IC50<1000 nM, which corresponds to the concentration of the test molecule that inhibits 50% of the competitive HLA-DP4 binding of the labeled peptide.Type: GrantFiled: October 17, 2002Date of Patent: May 18, 2010Assignees: Commissariat a l'Energie Atomique, Sedac TherapeuticsInventors: Bernard Maillere, Florence Castelli, Cécile Buhot, Bertrand Georges
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Patent number: 7713915Abstract: A method is provided for constructing, identifying and using new therapeutic or diagnostic proteins capable of binding to a target cell. The new proteins are derived by mutating a binding subunit of a wild type heteromeric cytotoxic protein to create a library of microorganism clones producing mutant proteins which are then screened for their ability to specifically bind to and kill a target cell.Type: GrantFiled: December 8, 1998Date of Patent: May 11, 2010Assignee: Molecular Templates, Inc.Inventors: Jean Gariepy, Mark Robert Bray
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Publication number: 20100113283Abstract: The present invention provides multiplexed methods for analyzing polynucleotides associated with sample tags. The multiplexed information is deconvoluted by single-molecule and more generally single-particle detection methods. In particular, a method for determining nucleic acid sequence information is provided.Type: ApplicationFiled: December 4, 2009Publication date: May 6, 2010Inventor: Michael Paul Strathmann
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Patent number: 7704925Abstract: The present invention in one aspect relates to a method for synthesizing a bifunctional complex comprising a molecule and an identifier polynucleotide identifying at least some of the chemical entities which have participated in the synthesis of the molecule in accordance with the methods of the present invention. The invention also relates to a library of different bifunctional complexes. The library of the invention can be used e.g. for identifying drug leads. Furthermore, the present invention is based on the principle that chemical entities initially provided on a building block oligonucleotide (i.e. a building block having an oligonucleotide part which is linked to a chemical entity) can be brought into reactive proximity without the use of a template comprising a set of covalently linked codons.Type: GrantFiled: March 22, 2005Date of Patent: April 27, 2010Assignee: Nuevolution A/SInventors: Alex Haahr Gouliaev, Henrik Pedersen
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Publication number: 20100081576Abstract: A method of sample analysis is provided. In certain embodiments, the method may comprise: a) contacting under primer extension conditions a genomic sample comprising a test genome with a set of at least ten sequence-specific primers that are complementary to sites in only one strand of a reference chromosomal region, to produce primer extension products, and b) analyzing the primer extension products to identify a chromosomal rearrangement in the test genome.Type: ApplicationFiled: October 1, 2008Publication date: April 1, 2010Inventors: Robert A. Ach, Carsten Carstens, Bernd Buehler
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Publication number: 20100075858Abstract: The invention provides coding compositions comprising mixtures of coding oligonucleotides and methods of using such compositions to code samples. The compositions and methods are useful for identifying, verifying, or authenticating any type of sample, whether the sample is biological or non-biological.Type: ApplicationFiled: May 22, 2009Publication date: March 25, 2010Applicant: GENVAULT CORPORATIONInventors: James C. DAVIS, Mitchell D. EGGERS, Rafael IBARRA, John SADLER, David WONG, Syrus M. JAFFE, Michael SAGHBINI, Michael HOGAN
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Publication number: 20100069250Abstract: Disclosed is a method for accurately determining the number of template molecules in a library of nucleic acids (e.g., DNA) to be sequenced. The method does not require large amounts of the DNA sample, nor does it require the preparation of a standard curve. The method is especially applicable to methodologies for “sequencing by synthesis,” where quantitation of the starting library is important. The method uses quantitative real time PCR, especially digital PCR, which measures the number of individual molecules in a sample. The present method particularly may use a microfluidic device for running large numbers of PCR reactions. Each PCR reaction is monitored in real time by a primer/probe combination. The forward primer is adapted to contain a sequence not on the adapter but which corresponds to a probe sequence. A short probe which generates fluorescence during the PCR process is used.Type: ApplicationFiled: August 14, 2009Publication date: March 18, 2010Applicant: The Board of Trustees of the Leland Stanford Junior UniversityInventors: Richard Allen White, III, Stephen R. Quake, Hei-Mun Christina Fan, Paul Blainey
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Patent number: 7666817Abstract: The present invention provides compositions including peptide display scaffolds that present at least one candidate peptide and at least one detectable moiety in at least one of the N-terminal and C-terminal candidate peptide presenting domains that when expressed in a cell are accessible at a surface of the cell outermembrane. In addition, the present invention also provides kits and methods for screening a library of cells presenting the candidate peptides in peptide display scaffolds to identify a ligand for an enzyme.Type: GrantFiled: August 30, 2006Date of Patent: February 23, 2010Assignee: The Regents of the University of CaliforniaInventors: Patrick Sean Daugherty, Kevin T. Boulware
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Publication number: 20100016170Abstract: A method for identifying gene cluster is disclosed. The method may be used for identifying gene clusters involved in the biosynthesis of natural products. A small insert library of DNA fragments of genomic DNA and a large insert library of DNA fragments of genomic DNA are prepared. Fragments in the small insert library are sequenced and compared by homology comparison under computer control to a database containing genes, gene fragments or proteins known to be involved in the biosynthesis of microbial natural products. Fragments having similar structure to genes, gene fragments or proteins known to be involved in the biosynthesis of naturally occurring metabolites are used as probes to screen the large insert library of genomic DNA to detect gene clusters involved in the biosynthesis of microbial natural products.Type: ApplicationFiled: May 14, 2007Publication date: January 21, 2010Applicant: Thallion Pharmaceuticals, Inc.Inventors: Chris M. Farnet, Alfredo Staffa, Emmanuel Zazopoulos
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Publication number: 20090318299Abstract: A method for high throughput screening of probes is described. These probes are useful for characterization and measurement of unbound metabolites in a fluid sample, particularly characterization and measurement of levels of unbound free fatty acids. By practice of the disclosed invention, a profile of unbound metabolites can be determined for an individual which can be used to determine the individual's relative risk for disease such as stroke, cardiac disease and cancer.Type: ApplicationFiled: August 4, 2009Publication date: December 24, 2009Applicant: FFA Sciences LLCInventors: ALAN MARC KLEINFELD, ANDREW HENRY HUBER, JAMES PATRICK KAMPF, THOMAS KWAN, BAOLONG ZHU
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Publication number: 20090286688Abstract: Methods disclosed herein relate to identification of nucleotides in a nucleotide sequence.Type: ApplicationFiled: July 20, 2009Publication date: November 19, 2009Inventors: Mark Chee, David R. Walt
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Publication number: 20090280993Abstract: A method of analyzing an accuracy of a sequence of a probe nucleic acid immobilized in a microarray includes providing a microarray in which regions where a probe nucleic acid is immobilized on a substrate are arrayed, hybridizing the probe nucleic acid with a target nucleic acid that is complementary to the probe nucleic acid to form a hybridization product, wherein the target nucleic acid is labeled with a detectable signal material on at least one end, reacting the hybridization product with an enzyme to remove the detectable signal material from the target nucleic acid, wherein the at least one end of the target nucleic acid remains unpaired with the probe nucleic acid, measuring a residual signal generated from the resultant enzyme reaction product, comparing the measured signal value with a signal value generated from a control group experiment; and analyzing an accuracy of the probe nucleic acid sequence.Type: ApplicationFiled: February 13, 2009Publication date: November 12, 2009Inventor: Joowon Rhee
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Publication number: 20090264300Abstract: Disclosed is a method for obtaining a bifunctional complex comprising a molecule linked to a single stranded identifier oligonucleotide, wherein a nascent bifunctional complex comprising a chemical reaction site and a priming site for enzymatic addition of a tag is a) reacted at the chemical reaction site with one or more reactants, and b) reacted enzymatically at the priming site with one or more tag(s) identifying the reactant(s).Type: ApplicationFiled: December 1, 2006Publication date: October 22, 2009Applicant: Nuevolution A/SInventors: Thomas Franch, Mikkel Dybro Lundorf, Soren Nyboe Jacobsen, Eva Kampmann Olsen, Anne Lee Andersen, Anette Holtmann, Anders Holm Hansen, Anders Malling Sorensen, Anne Goldbech, Daen de Leon, Ditte Kievsmose Kaldor, Frank Abildgaard Slok, Gitte Nystrup Husemoen, Johannes Dolberg, Kim Birkebaek Jensen, Lene Pedersen, Mads Norregaard-Madsen, Michael Anders Godskesen, Sanne Schroder Glad, Sore Neve, Thomas Thisted, Tine Titilola Akinleminu Kronborg, Christian Klarner Sams, Jakob Felding, Per-Ola Freskgard, Alex Haahr Gouliaev, Henrik Pedersen
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Publication number: 20090233855Abstract: Compositions and methods for modulating the activation of nuclear factor ?B (NF-?B) are provided. The compositions comprise one or more agents that modulate ubiquitination of phosphorylated I?B? and/or I?B? Such compositions may be used for treating diseases associated with NF-?B activation. Modulating agents include human E3 ubiquitin ligases, antibodies thereto and variants thereof, as well as related proteins.Type: ApplicationFiled: August 8, 2008Publication date: September 17, 2009Inventors: Anthony M. MANNING, Frank Mercurio, Sharon Amit, Yinon Ben-Neriah, Matti Davis, Ada Hatzubai, Avraham Yaron, Irit Alkalay, Aaron Ciechanover
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Patent number: 7585815Abstract: Methods, compositions, and kits for the cell-free production and analysis of proteins are provided. The invention allows for the production of proteins from prokaryotic sequences or eukaryotic sequences, including human cDNAs using PCR and IVT methods and detecting the proteins through fluorescence or immunoblot techniques. This invention can be used to identify optimized PCR and WT conditions, codon usages and mutations. The methods are readily automated and can be used for high throughput analysis of protein expression levels, interactions, and functional states.Type: GrantFiled: July 1, 2004Date of Patent: September 8, 2009Assignee: Lawrence Livermore National Security, LLCInventors: Peter T. Beernink, Matthew A. Coleman, Brent W. Segelke
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Publication number: 20090221428Abstract: The invention relates to improved methods of identifying the genomic regions to which a protein of interest binds, and in particular, to methods that apply to stem cells such as but not limited to; embryonic stem cells and adult stem cells. The invention also provides methods of identifying agents which modulate differentiation of stem cells. The invention also provides methods of defining the differentiation potential of a cell and of designing array oligonucleotides.Type: ApplicationFiled: September 7, 2006Publication date: September 3, 2009Inventors: Richard A. Young, Tong Ihn Lee, Matthew Guenther, Laurie A. Boyer
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Publication number: 20090221429Abstract: It is intended to provide a method for detecting a target nucleic acid with a specific base sequence existing in a sample mixture with high specificity and sensitivity by utilizing the formation of a hybrid with a complementary strand as a detection principle, and a nucleic acid for the detection.Type: ApplicationFiled: November 17, 2006Publication date: September 3, 2009Applicant: JAPAN SCIENCE AND TECHNOLOGY AGENCYInventors: Kenzo Fujimoto, Yoshinaga Yoshimura, Shinzi Ogasawara
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Publication number: 20090215633Abstract: Method for the detection the presence or absence of one or more target sequences in one or more samples based on oligonucleotide ligation assays with a variety of ligatable probes containing identifiers and the subsequent identification of the identifiers in the amplicons or ligated probes using high throughput sequencing technologies.Type: ApplicationFiled: March 1, 2007Publication date: August 27, 2009Inventors: Michael Josephus Theresia Van Eijk, Rene Cornelis Josephus Hogers
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Publication number: 20090209430Abstract: The present invention provides a method for combining the advantages of encoded molecule fragments made by split and mix synthesis with the advantages of template directed synthesis of molecules.Type: ApplicationFiled: November 22, 2005Publication date: August 20, 2009Inventor: Peter Birk Rasmussen
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Patent number: 7566685Abstract: The invention relates to methods and compositions utilizing diphtheria toxin for screening purposes. The invention is particularly useful in screening for modulators of IgE synthesis, secretion and switch rearrangement.Type: GrantFiled: March 28, 2006Date of Patent: July 28, 2009Assignee: Rigel Pharmacauticals, Inc.Inventor: Todd M. Kinsella