Directed Molecular Evolution Of Macromolecules (e.g., Rna, Dna, Proteins, Etc.) Patents (Class 506/1)
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Publication number: 20140302998Abstract: The present application provides stable peptide-based Akt capture agents and the use thereof as detection, diagnosis, and treatment agents. The application further provides novel methods of developing stable peptide-based capture agents, including Akt capture agents, using iterative on-bead in situ click chemistry.Type: ApplicationFiled: July 11, 2012Publication date: October 9, 2014Inventors: James R. Heath, Arundhati Nag, Samir Das, Kaycie M. Deyle, Steven Wesley Millward, Paul Edward Kearney
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Patent number: 8835162Abstract: The invention provides a method of incorporating nonstandard amino acids into a protein by utilizing a modified aminoacyl-tRNA synthetase to charge the nonstandard amino acid to a modified tRNA, which forms strict Watson-Crick base-pairing with a codon that normally forms wobble base-pairing with natural tRNAs.Type: GrantFiled: December 28, 2012Date of Patent: September 16, 2014Assignee: California Institute of TechnologyInventors: Inchan Kwon, David Tirrell
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Publication number: 20140256557Abstract: The disclosure relates to a method of generating a diverse set of variants to screen improved and novel properties within the variant population, a system for creating the diverse set of variants, and the variant peptides.Type: ApplicationFiled: May 19, 2014Publication date: September 11, 2014Applicant: Codexis, Inc.Inventor: Richard Fox
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Publication number: 20140249035Abstract: The present invention generally relates to methods of rapidly and efficiently searching biologically-related data space. More specifically, the invention includes methods of identifying bio-molecules with desired properties, or which are most suitable for acquiring such properties, from complex bio-molecule libraries or sets of such libraries. The invention also provides methods of modeling sequence-activity relationships. As many of the methods are computer-implemented, the invention additionally provides digital systems and software for performing these methods.Type: ApplicationFiled: April 18, 2014Publication date: September 4, 2014Applicant: Codexis Mayflower Holdings, LLCInventor: Richard John Fox
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Publication number: 20140243209Abstract: The present invention provides a method for detecting an interaction, which method can solve not only the problem of false negatives but also the problem of false positives.Type: ApplicationFiled: September 28, 2012Publication date: August 28, 2014Applicant: The University of TokyoInventors: Etsuko Miyamoto, Shigeo Fujimori
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Publication number: 20140243208Abstract: The invention encompasses compositions and methods for selecting aptamers.Type: ApplicationFiled: July 23, 2012Publication date: August 28, 2014Applicant: Mediomics, LLCInventors: Yie-Hwa Chang, Ling Tian, Rongsheng Wang
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Publication number: 20140221216Abstract: The present invention provides methods for identifying bio-molecules with desired properties, or which are most suitable for acquiring such properties, from complex bio-molecule libraries or sets of such libraries. More specifically, some embodiments of the present invention provide methods for building sequence-activity models comprising multiplicative terms and using the models to guide directed evolution. In some embodiments, the sequence-activity models include one or more interaction terms, each of which including an interaction coefficient representing the contribution to activity of two or more defined residues. In some embodiments, the models describe relation between protein or nucleic acid sequences and protein activities. In some embodiments, the present invention also provides methods for preparing sequence-activity models, including but not limited to stepwise addition or subtraction techniques, Bayesian regression, ensemble regression and other methods.Type: ApplicationFiled: January 29, 2014Publication date: August 7, 2014Inventors: Gregory Allan Cope, Nicholas John Agard
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Publication number: 20140213460Abstract: Methods relating to obtaining libraries of YY1-binding long non-coding RNAs, libraries obtained thereby, and methods of use thereof.Type: ApplicationFiled: July 3, 2012Publication date: July 31, 2014Applicant: THE GENERAL HOSPITAL CORPORATIONInventors: Jeannie T. Lee, Yesu Jeon
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Publication number: 20140213459Abstract: The present invention relates to methods for improving the folding stability of antibodies, to antibodies with improved folding stability, nucleic acid and vectors encoding such antibodies, and to uses of such antibodies, nucleic acid and vectors.Type: ApplicationFiled: May 29, 2012Publication date: July 31, 2014Inventor: Roland Beckmann
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Publication number: 20140206846Abstract: The present invention relates to antibody-based dual targeting molecules, and to methods for generating such dual targeting molecules, including a library-based approach.Type: ApplicationFiled: May 29, 2012Publication date: July 24, 2014Inventor: Roland Beckmann
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Publication number: 20140200145Abstract: The invention relates to a method for metabolic evolution of a variant of a natural small aromatic molecule product of a metabolic pathway, by somatic in vivo assembly and recombination of said metabolic pathway employing a gene mosaic of at least one gene A, which comprises a) in a single step procedure (i) transforming a cell with at least one gene A having a sequence homology of less than 99.Type: ApplicationFiled: June 20, 2012Publication date: July 17, 2014Applicant: Eviagenics S.A.Inventors: Rudy Pandjaitan, Sarra Sebai, Alejándro Luque
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Patent number: 8759058Abstract: The present invention relates generally to hydrogen production for use in fuel cells, foodstuffs and chemical production, and more particularly, to biologically and photosynthetically produced hydrogen. Specifically, disclosed is a method for producing bacteria and green alga that can produce hydrogen in quantities that exceed four hundred percent of the hydrogen produced by green alga in nature; thus, producing organisms which can serve as hydrogen generators for fuel cells, chemical production and numerous other applications.Type: GrantFiled: February 24, 2012Date of Patent: June 24, 2014Inventors: Scott Plummer, Mark Plummer
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Patent number: 8741810Abstract: The present invention relates to a high efficiency method of expressing immunoglobulin molecules in eukaryotic cells. The invention is further drawn to a method of producing immunoglobulin heavy and light chain libraries, particularly using the trimolecular recombination method, for expression in eukaryotic cells. The invention further provides methods of selecting and screening for antigen-specific immunoglobulin molecules, and antigen-specific fragments thereof. The invention also provides kits for producing, screening and selecting antigen-specific immunoglobulin molecules. Finally, the invention provides immunoglobulin molecules, and antigen-specific fragments thereof, produced by the methods provided herein.Type: GrantFiled: August 17, 2007Date of Patent: June 3, 2014Assignee: University of RochesterInventors: Maurice Zauderer, Ernest S. Smith
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Publication number: 20140121115Abstract: New rare-cutting endonucleases, also called custom-made meganucleases, which recognize and cleave a specific nucleotide sequence, derived polynucleotide sequences, recombinant vector cell, animal, or plant comprising said polynucleotide sequences, process for producing said rare-cutting endonucleases and any use thereof, more particularly, for genetic engineering, antiviral therapy and gene therapy.Type: ApplicationFiled: December 21, 2012Publication date: May 1, 2014Applicant: CellectisInventors: Sylvain Arnould, Sylvia Bruneau, Jean-Pierre Cabaniols, Patrick Chames, Andre Choulika, Phillipe Duchateau, Jean-Charles Epinat, Agnes Gouble, Emmanuel Lacroix, Frederic Paques, Christophe Perez-Michaut, Julianne Smith, Davie Sourdive
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Publication number: 20140109249Abstract: The present invention relates to methods for the screening, identification and/or application of microorganisms of use in imparting beneficial properties to plants.Type: ApplicationFiled: October 10, 2013Publication date: April 17, 2014Applicant: Biodiscovery New Zealand LimitedInventors: Susan Jane Turner, Peter John Wigley
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Publication number: 20140100120Abstract: Provided herein are methods for a novel bead-based next-generation “X-aptamer” selection scheme that extends aptamer technology to include X-modified bases, thus resulting in X-aptamers, at any position along the sequence because the aptamers are chemically synthesized via a split-pool scheme on individual beads. Also provides are application to a wide range of commonly used DNA modifications, including, but not limited to, monothioate and dithioate backbone substitutions. This new class of aptamer allows chemical modifications introduced to any of the bases in the aptamer sequence as well as the phosphate backbones and can be extended to other carbohydrate-based systems.Type: ApplicationFiled: October 10, 2013Publication date: April 10, 2014Inventors: David G Gorenstein, Weiguo He, David E Volk, Miquel-Angel Elizondo-Riojas, Ross Durland, Johnnie Engelhardt
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Publication number: 20140094373Abstract: The invention provides methods for simultaneously amplifying multiple nucleic acid regions of interest in one reaction volume as well as methods for selecting a library of primers for use in such amplification methods. The invention also provides library of primers with desirable characteristics, such as minimal formation of amplified primer dimers or other non-target amplicons.Type: ApplicationFiled: October 2, 2013Publication date: April 3, 2014Applicant: Natera, Inc.Inventors: Berhard Zimmermann, Matthew Hill, Philippe Lacroute, Michael Dodd
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Publication number: 20140056959Abstract: The present invention relates to optimized aptamers and methods of using these aptamers.Type: ApplicationFiled: August 12, 2013Publication date: February 27, 2014Applicant: University of Iowa Research FoundationInventors: Paloma Giangrande, Francis Miller, William Thiel
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Publication number: 20130345064Abstract: The invention provides systems, methods, reagents, apparatuses, vectors, and host cells for the continuous evolution of nucleic acids. For example, a lagoon is provided in which a population of viral vectors comprising a gene of interest replicates in a stream of host cells, wherein the viral vectors lack a gene encoding a protein required for the generation of infectious viral particles, and wherein that gene is expressed in the host cells under the control of a conditional promoter, the activity of which depends on a function of the gene of interest to be evolved. Some aspects of this invention provide evolved products obtained from continuous evolution procedures described herein. Kits containing materials for continuous evolution are also provided.Type: ApplicationFiled: December 22, 2011Publication date: December 26, 2013Applicant: President and Fellows of Harvard CollegeInventors: David R. Liu, Kevin Michael Esvelt, Jacob Charles Carlson
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Publication number: 20130316910Abstract: An object of the invention is to provide a peptide having a stabilized secondary structure. The present invention provides a peptide having a secondary structure stabilized by a crosslinked structure and containing at least one combination of a special amino acid of the formula (I): (wherein, (A) represents a single bond or a linking group having, in the main chain thereof, from 1 to 10 atoms; (B) represents a group containing at least one ? bond; (C) represents a hydrogen atom or an alkyl group which may be substituted with a substituent; and X represents a group substitutable by a substitution reaction with a sulfanyl group) and an amino acid having, in the side chain thereof, a sulfanyl group; and having the crosslinked structure formed through a thioether bond between the side chain of the special amino acid residue and the sulfanyl group.Type: ApplicationFiled: December 5, 2011Publication date: November 28, 2013Applicant: The University of TokyoInventors: Hiroaki Suga, Takashi Higuchi
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Publication number: 20130281303Abstract: The present invention relates to methods for efficiently generating recombinant monoclonal antibodies derived from B cells of a non-human host which has been immunochallenged with one or more target antigens. The methods comprise the steps of identifying and isolating B cell that bind to the antigen by FACS, and recombining and enriching for thousands of cells to create a B cell library. Related products and methods, such as methods of producing expression libraries, are also disclosed.Type: ApplicationFiled: December 28, 2011Publication date: October 24, 2013Applicant: BioAtla, LLCInventor: Jay M. Short
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Publication number: 20130274112Abstract: Disclosed is a method that includes: (i) providing a plurality of initial nucleic acid cassettes that include: a) a first coding region encoding a first immunoglobulin variable domain, b) a second coding region encoding a second immunoglobulin variable domain, and c) a ribosomal binding site disposed between the first and second coding regions for translation of the second polypeptide in a first expression system, wherein the first and second coding regions are in the same translational orientation; (ii) modifying each nucleic acid cassette of the plurality in a single reaction mixture so that it is functional in a second expression system, wherein the first and second region remain physically attached during the modifying; (iii) introducing each modified nucleic acid cassette into a mammalian cell to produce a mixture of transfected cells; and (iv) expressing each modified nucleic acid cassette in the transfected cells.Type: ApplicationFiled: April 22, 2013Publication date: October 17, 2013Inventors: Henricus R J. Hoogenboom, Jurgen Mullberg, Robert C. Ladner
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Publication number: 20130274113Abstract: The present invention relates to a multiplex microfluidic device for selection of nucleic acid aptamers and a method for high-throughput selection of nucleic acid aptamers using the same, and more particularly to a multiplex microfluidic device (SELEX lap-on-a-chip) that uses an improved multiplex platform in place of the development of an aptamer for a single target and to a method for high throughput selection of aptamers using the same together with high-throughput sequencing. A multiplex microfluidic device according to the present invention can simultaneously detect aptamers for a plurality of targets, and it can greatly increase the screening throughput and greatly shorten the process time compared to conventional multiplex techniques.Type: ApplicationFiled: October 5, 2011Publication date: October 17, 2013Applicant: PCL, INC.Inventors: So Youn Kim, Ji-Young Ahn, Minjoung Jo, Tae Kyung Kim
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Publication number: 20130267676Abstract: The present invention provides a fibronectin type III (Fn3) molecule, wherein the Fn3 contains a stabilizing mutation. The present invention also provides Fn3 polypeptide monobodies, nucleic acid molecules encoding monobodies, and variegated nucleic acid libraries encoding such monobodies. Also provided are methods of preparing a Fn3 polypeptide monobody, and kits to perform the methods.Type: ApplicationFiled: August 2, 2012Publication date: October 10, 2013Inventor: Shohei Koide
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Publication number: 20130266516Abstract: The present invention relates to an aptamer comprising a nucleic acid comprising, or consisting of: the sequence ACUGU CCCAG UAUGA CGCGA CUGCU UAGGU GGGAU GUUUC CCAUG CCUCG (SEQ ID NO: 1), or a sequence comprising, or consisting of, at least 25 consecutive nucleotides in a sequence having at least 80% identity with SEQ ID NO: 1, with the proviso that a nucleic acid consisting of this sequence binds to the LAR protein.Type: ApplicationFiled: December 15, 2011Publication date: October 10, 2013Applicant: COMMISSARIAT A L'ENERGIE ATOMIQUE ET AUX ENERGIESInventors: Frederic Duconge, Agnes Cibiel
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Publication number: 20130244245Abstract: The invention relates to orthogonal pairs of tRNAs and aminoacyl-tRNA synthetase that can incorporate unnatural amino acid into proteins produced in eubacterial host cells such as E. coli, or in a eukaryotic host such as a yeast cell. The invention provides, for example but not limited to, novel orthogonal synthetases, methods for identifying and making the novel synthetases, methods for producing proteins containing unnatural amino acids, and translation systems.Type: ApplicationFiled: June 8, 2012Publication date: September 19, 2013Applicant: The Scripps Research InstituteInventors: Peter G. Schultz, Lital Alfonta, Johnathan R. Chittuluru, Alexander Deiters, Dan Groff, Daniel Summerer, Meng-Lin Tsao, Jiangyun Wang, Ning Wu, Jianming Xie, Huaqiang Zeng, Mohammad Seyedsayamdost, James Turner
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Patent number: 8525533Abstract: A conductivity detector detects the electrical conductivity of a fluid under analysis for determining chemical or physical properties of the fluid that are related its electrical properties. Such conductivity detectors may find use in, for example, hemodialysis systems for analyzing the effectiveness of the hemodialysis treatment. In an aspect, to improve accuracy of the conductivity measurements, the detector utilizes four-wire resistance measurement methods. In another aspect, to avoid fouling or contamination of the electrodes, the detector utilizes capacitively-coupled contactless conductivity detection (C4D) methods so that the electrodes are physically unconnected to the fluid contained in a fluid chamber. In a possible further aspect, the fluid chamber may be a disposable component removable from the electrodes. The conductivity detector can include other features such as calibration circuits and features for electrically isolating the fluid under detection from the fluid in the rest of the system.Type: GrantFiled: September 16, 2010Date of Patent: September 3, 2013Assignee: Fresenius Medical Care Holdings, Inc.Inventor: Thomas A. Sullivan
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Publication number: 20130190189Abstract: The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising of the steps of: (a) compartmentalising genetic elements into microcapsules; (b) expressing the genetic elements to produce their respective gene products within the microcapsules; (c) sorting the genetic elements which produce the gene product having a desired activity. The invention enables the in vitro evolution of nucleic acids by repeated mutagenesis and iterative applications of the method of the invention.Type: ApplicationFiled: January 3, 2013Publication date: July 25, 2013Applicant: Medical Research CouncilInventor: Medical Research Council
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Publication number: 20130184160Abstract: The present disclosure encompasses oligonucleotide aptamers selectively binding a target glycosylated polypeptide or protein, and having biased affinity for the glycan through a boronic acid linked to a nucleosidic base of a nucleotide(s). The disclosure further encompasses methods for isolating an aptamer(s) selectively binding a target glycosylated polypeptide, where, from a population of randomized oligonucleotides that have at least one nucleotide having a boronic acid label linked to a base, is selected a first subpopulation of aptamers binding to the target glycosylated polypeptide or protein. This subpopulation is then amplified without using boronic acid-modified TTP, and amplification products not binding to a target glycosylated polypeptide or protein are selected. The second subpopulation of aptamers is then amplified using boronic acid-modified TTP to provide a population of boronic acid-modified aptamers capable of selectively binding to a glycosylation site of a target polypeptide or protein.Type: ApplicationFiled: July 17, 2008Publication date: July 18, 2013Inventors: Binghe Wang, Minyong Li, Zhen Huang, Na Lin
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Publication number: 20130178368Abstract: The invention describes a method for isolating one or more genetic elements encoding a gene product having a desired activity, comprising the steps of: (a) compartmentalising genetic elements into microcapsules; and (b) sorting the genetic elements which express the gene product having the desired activity; wherein at least one step is under microfluidic control. The invention enables the in vitro evolution of nucleic acids and proteins by repeated mutagenesis and iterative applications of the method of the invention.Type: ApplicationFiled: December 5, 2012Publication date: July 11, 2013Inventors: Andrew David Griffiths, David Weitz, Darren Link, Keunho Ahn, Jerome Bibette
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Publication number: 20130164271Abstract: The present invention relates to a method for preparing an expression vector encoding a tailored recombinase, which tailored recombinase is capable of recombining asymmetric target sequences within the long terminal repeat (LTR) of proviral DNA of a plurality of retrovirus strains inserted into the genome of a host cell, as well as to the obtained expression vector, cells transfected with this, expressed recombinase and pharmaceutical compositions comprising the expression vector, cells and/or recombinase. Pharmaceutical compositions are useful, e.g., in treatment and/or prevention of retrovirus infection. In particular, asymmetric target sequences present in a plurality of HIV strains are disclosed, as well as tailored recombinases capable of combining these sequences (Tre 3.0 and 4.0) and expression vectors encoding them.Type: ApplicationFiled: May 27, 2011Publication date: June 27, 2013Applicants: MAX-PLANCK-GESELLSCHAFT ZUR FOEDERUG DER WISSENSCH E.V., HENRICH-PETTE-INSTITUT LEIBNZ-INSTITUT FOR EXPERIM VIROLOGIE-STIFTUNG BInventors: Joachim Hauber, Jan Chemnitz, Frank Buchholz, Janet Chusainow
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Patent number: 8470164Abstract: Methods and apparatus relating to very large scale FET arrays for analyte measurements. ChemFET (e.g., ISFET) arrays may be fabricated using conventional CMOS processing techniques based on improved FET pixel and array designs that increase measurement sensitivity and accuracy, and at the same time facilitate significantly small pixel sizes and dense arrays. Improved array control techniques provide for rapid data acquisition from large and dense arrays. Such arrays may be employed to detect a presence and/or concentration changes of various analyte types in a wide variety of chemical and/or biological processes. In one example, chemFET arrays facilitate DNA sequencing techniques based on monitoring changes in the concentration of inorganic pyrophosphate (PPi), hydrogen ions, and nucleotide triphosphates.Type: GrantFiled: June 25, 2009Date of Patent: June 25, 2013Assignee: Life Technologies CorporationInventors: Jonathan Rothberg, Wolfgang Hinz, Kim Johnson, James Bustillo, John Leamon, Jonathan Schultz
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Publication number: 20130137582Abstract: A method for determining an efficiency of target enrichment from a DNA library, includes: adding a negative control sequence and a positive control sequence to the DNA library, or picking a negative control sequence and/or a positive control sequence from the library; determining a pre-capture amount of the negative control sequence and a pre-capture amount of the positive control sequence; performing enrichment of a target sequence from the DNA library using a bait sequence to produce a post-capture library; determining a post-capture amount of the negative control sequence and a post-capture amount of the positive control sequence in the post-capture library; and determining the efficiency of the target enrichment, based on the post-capture amount of the positive control sequence, the post-capture amount of the negative control sequence, the pre-capture amount of the positive control sequence, and the pre-capture amount of the negative control sequence.Type: ApplicationFiled: November 30, 2011Publication date: May 30, 2013Applicant: AGILENT TECHNOLOGIES, INC.Inventors: Joseph Hoang Hai Ong, Scott Happe
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Publication number: 20130116417Abstract: Described herein are oligosaccharide-oligonucleotide conjugates useful as vaccines against one or more human or veterinary therapeutic indications, and methods of synthesizing and identifying them. The conjugates may be identified using non-human antibodies as binding targets, thereby expanding the power and scope of the invention. Efficacious conjugates may be identified through an iterative.Type: ApplicationFiled: June 10, 2011Publication date: May 9, 2013Applicant: BRANDEIS UNIVERSITYInventors: Isaac J. Krauss, Lizbeth K. Hedstrom, Iain S. MacPherson
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Publication number: 20130116125Abstract: The present invention is relevant to proteins and novel methods of protein evolution. The present invention further relates to methods of identifying and mapping mutant polypeptides formed from, or based upon, a template polypeptide.Type: ApplicationFiled: March 4, 2011Publication date: May 9, 2013Applicant: BIOATLA, LLCInventor: Jay M. Short
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Publication number: 20130090246Abstract: The invention relates to a method for generating a gene mosaic by somatic in vivo recombination, comprising: e) in a single step procedure (vii) transforming a cell with at least one gene A having a sequence homology of less than 99.5% to another gene to be recombined that is an integral part of the cell genome or presented in the framework of a genetic construct, (viii) recombining said genes, (ix) generating a gene mosaic of the genes at an integration site of a target genome, wherein said at least one gene A has a single flanking target sequence either at the 5? end or 3? end anchoring to the 5? or 3? end of said integration site, and f) selecting clones comprising the gene mosaic, as well as a method of producing a diversity of gene mosaics and gene assembly.Type: ApplicationFiled: April 8, 2011Publication date: April 11, 2013Applicant: EVIAGENICS S.A.Inventors: Rudy Pandjaitan, Alejandro Luque
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Publication number: 20130085072Abstract: Described herein are methods that combine phage and yeast display to create polyclonal antibodies that are renewable, and when amplified over 100 million fold, maintain diversity without loss of representation of any of the antibodies present. The antibody representation remains essentially constant, as confirmed by deep sequencing. The provided methods allow generation, use and propagation of polyclonal antibodies, without concern that representation is lost. Furthermore, because the derivation of the polyclonal pool is carried out in vitro using phage and yeast display, it is possible in various embodiments to eliminate reactivities that are considered undesirable. Additionally, the polyclonal pool can be enriched for higher affinity antibodies.Type: ApplicationFiled: September 28, 2012Publication date: April 4, 2013Applicant: LOS ALAMOS NATIONAL SECURITY, LLCInventor: LOS ALAMOS NATIONAL SECURITY, LLC
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Publication number: 20130059732Abstract: Recombinant adeno-associated viral (AAV) capsid proteins are provided. Methods for generating the recombinant adeno-associated viral capsid proteins and a library from which the capsids are selected are also provided.Type: ApplicationFiled: August 24, 2012Publication date: March 7, 2013Applicant: THE BOARD OF TRUSTEES OF THE LELAND STANFORD JUNIOR UNIVERSITYInventors: Leszek Lisowski, Mark A. Kay
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Publication number: 20130053543Abstract: A peptide clearing agent is provided for clearance of a conjugate of an enzyme and a binding molecule which binds specifically at a target location from a non-target location in a subject. The peptide clearing agent binds the active site of the enzyme. The peptide also binds to the asialoglycoprotein receptor expressed by hepatic cells to facilitate clearance through the liver. The peptide may be glycosylated to facilitate clearance through the liver by binding to hepatic cells expressing an asialoglyco-protein receptor. Typically, the peptide prevents or inhibits enzyme activity upon binding to the enzyme and is not substantially modified by the enzyme activity. The peptide may be based upon the dipeptide amino-naphthoic acid (ANA)-glutamate (GIu) and may comprise the amino acid sequence serine (Ser)-Alanine (Ala)-amino-naphthoic acid (ANA)-glutamate (GIu). In such cases, the enzyme of interest is typically CPG2.Type: ApplicationFiled: September 23, 2010Publication date: February 28, 2013Applicant: MOLOGIC LTDInventors: Paul James Davis, James Alexander Schouten
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Publication number: 20130045871Abstract: The present invention provides systems, vectors and methods for isolation of enhanced ligand-binding proteins from combinatorial libraries displayed on the inner membrane of a host cell.Type: ApplicationFiled: March 18, 2011Publication date: February 21, 2013Applicant: CORNELL UNIVERSITYInventors: Matthew DeLisa, Hyung-Kwon Lim, Amy Karlsson
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Publication number: 20130040823Abstract: Disclosed is a method for obtaining a bifunctional complex comprising a display molecule part and a coding part, wherein a nascent bifuntional complex comprising a chemical reaction site and a priming site for enzymatic addition of a tag is reacted at the chemical reaction site with one or more reactants, and provided with respective tag(s) identifying the reactant(s) at the priming site is using one or more enzymes.Type: ApplicationFiled: April 25, 2012Publication date: February 14, 2013Applicant: NUEVOLUTION A/SInventors: Per-Ola FRESKGARD, Thomas FRANCH, Alex Haahr GOULIAEV, Mikkel Dybro LUNDORF, Jakob FELDING, Eva Kampmann OLSEN, Anette HOLTMANN, Soeren Nyboe JAKOBSEN, Christian Klarner SAMS, Sanne Schroeder GLAD, Kim Birkebaek JENSEN, Henrik PEDERSEN
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Publication number: 20130034879Abstract: A DNA polymerase mutant comprising a Taq DNA polymerase amino acid sequence with a mutation at one or more of the following selected amino acid positions: E189K, E230K, E507K, H28R, L30R, G38R, F73V, H75R, E76A, E76G, E76K, E90K, K206R, E315K, A348V, L351F, A439T, D452N, G504S, E507A, D551N, L552R, I553V, D578N, H676R, Q680R, D732G, E734G, E734K, F749V; wherein the polymerase mutant exhibits relative to wild-type DNA polymerase increased polymerase speed, increased affinity to DNA substrate and/or increased resistance to a DNA polymerase inhibitor; and wherein, when the mutation is E507K in combination with two or more further mutations or the mutation is Q680R in combination with four or more further mutations, at least one of the further mutations is at one of the selected amino acid positions; and when the mutation is I553V, this is not in combination with D551S.Type: ApplicationFiled: August 2, 2012Publication date: February 7, 2013Applicant: Fermentas UABInventors: Remigijus Skirgaila, Agne Tubeleviciute, Renata Rimseliéné, Sigitas Burinskas
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Publication number: 20130029850Abstract: The invention is drawn to a novel method useful for screening. In particular, the present invention provides methods for competitive differential screening. In some preferred embodiments, the present invention provides methods for competitive differential screening that facilitate the identification of tight binders. In some preferred embodiments, the agents used in the methods of the present invention comprise tight and weak binders. In other embodiments, the present invention provides methods that utilize competitive binders that recognize and bind targets, but with binding that is less strong than that of binders of interest.Type: ApplicationFiled: July 30, 2012Publication date: January 31, 2013Applicant: DANISCO US INC.Inventors: Anthony G. Day, David A. Estell, Paul T. Jedrzejewski, Christopher J. Murray
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Publication number: 20130005582Abstract: Method of preparation of restriction endonucleases, particularly those exhibiting the desired sequential specificity consists in that a fluorescence-marked DNA probe is used for screening a library of mutants, preferably in IVC format, and/or using other high-performance screening (HTS) technique, which is attained through expression of proteins included in the library of mutants in a cell-free system in the presence and by means of the DNA probe, and proteins thus obtained, resulting from expression of clones from the library, degrade the DNA probe, if their substrate specificity matches the searched one, the degradation of the DNA probe being detected as a disappearance of the FRET phenomenon between fluorescence markers included in the probe, and then microcompartments in which the FRET phenomenon ceases to occur, are separated from the remaining ones using Fluorescence Activated Cell Sorter (FACS) and/or other equipment for HTS analysis, and then DNA coding clones capable of degrading the probe are amplifType: ApplicationFiled: September 28, 2010Publication date: January 3, 2013Inventor: Michal Lower
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Publication number: 20130005581Abstract: Disclosed is a composition comprising the nucleic acid and a chemical compound, said composition forming a star structure defining 3 or more stems extending from a reaction center. The stems are formed by a nucleic acid duplex and the chemical compound has been formed in the reaction center as the reaction product of 3 or more chemical groups. The advantage of the composition is that a close proximity is provided between the chemical groups in the reaction center, thereby promoting a reaction. The invention also relates to a method for preparation of the composition. The advantage of the method is that it does not require the pre-synthesis of a large number of templates and that it is not dependent upon codon/anti-codon recognition for an encoded molecule to be formed.Type: ApplicationFiled: May 17, 2012Publication date: January 3, 2013Applicant: VIPERGEN APSInventors: Nils Jokob Vest Hansen, Peter Blakskjaer, Margit Haahr Hansen, Lars Kolster Petersen, Tara Renee Heitner
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Publication number: 20120329067Abstract: Provided herein are zinc linger nucleases having altered, arid in particular, improved catalytic activity and methods of generating such nucleases. Accordingly, there are provided methods for identifying improved catalytic activity of a ZFN by expressing a mutated zinc finger nuclease in a cell containing a reporter construct with a toxic gene, and a zinc finger nuclease cleavage site that is recognized by the ZFN. Survival of the cell is positively correlated with catalytic activity of the ZFN; thus, libraries of mutated ZFKs may be selected for altered catalytic activity based on relative survival rates, Methods of using identified ZFNs are also provided.Type: ApplicationFiled: January 21, 2011Publication date: December 27, 2012Inventors: Carlos F. Barbas III, Jing Guo
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Publication number: 20120316071Abstract: Provided herein is a rational method of affinity maturation to evolve the activity of an antibody or portion thereof based on the structure/affinity or activity relationship of an antibody. The resulting affinity matured antibodies exhibit improved or optimized binding affinity for a target antigen.Type: ApplicationFiled: November 4, 2010Publication date: December 13, 2012Inventors: Vaughn Smider, Helen Hongyuan Mao
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Publication number: 20120295793Abstract: The invention provides for methods of generating modified polynucleotide libraries by inserting and/or deleting at least three nucleotide residues in polynucleotide sequences. Theses methods may be used with other methods of gene modification such as gene shuffling. The invention further provides methods of directed molecular evolution using the modified polynucleotide libraries produced by these methods.Type: ApplicationFiled: January 21, 2011Publication date: November 22, 2012Applicant: ProteusInventors: Christophe Ullman, Laurent Fourage
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Publication number: 20120264616Abstract: The present invention concerns a method of directing evolution of a target polynucleotide of interest for obtaining variants of this target polynucleotide, as well as a method to generate genetic variability by preparing a cell library. This invention also relates to a method to isolate or to screen variants of a polynucleotide or variants of a protein able to impact the phenotype of a cell or to confer a desired phenotype to target cells, and to identify theses polynucleotide variants or protein variants responsible for this phenotype.Type: ApplicationFiled: November 10, 2010Publication date: October 18, 2012Applicants: UNIVERSITE PIERRE ET MARIE CURIE (PARIS 6), CENTRE NATIONAL DE LA RECHERCHE SCIENTIFIQUEInventors: Matteo Negroni, Sarah Gallois-Montbrun, Paola Rossolillo, Vincenzo Di Bartolo, Gilles Uze, Etienne Simon-Loriere, Roland Marquet, Valérie Vivet-Boudou
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Publication number: 20120258865Abstract: This disclosure relates to a method of generating conditionally active biologic proteins from wild type proteins, in particular therapeutic proteins, which are reversibly or irreversibly inactivated at the wild type normal physiological conditions. For example, evolved proteins are virtually inactive at body temperature, but are active at lower temperatures.Type: ApplicationFiled: June 14, 2012Publication date: October 11, 2012Applicant: BioAtla, LLCInventors: Jay M. Short, Hwai Wen Chang, Gerhard Frey