Abstract: Disclosed herein are methods for inducing production of full-length progranulin (GRN) from a nucleotide encoding a GRN with a premature stop codon (GRN-PTC), comprising exposing the GRN-PTC to an aminoglycoside. Methods for inducing production of full-length progranulin (GRN) from a nucleotide encoding a GRN with a premature stop codon (GRN-PTC), comprising administering gentamicin or G418 is also disclosed.

Abstract: Provided herein, in some embodiments, are methods of purifying a nucleic acid preparation. The methods may comprise contacting a nucleic acid preparation comprising messenger ribonucleic acid with an RNase III enzyme that is immobilized on a solid support and binds to double-stranded RNA contaminants.

Abstract: The invention relates a recombinant measles virus plasmid capable of expressing a human telomerase reverse transcriptase (hTERT) protein fused at N-terminus with a protein enhancing addressing of the hTERT protein to proteasome. The invention further relates to a vaccine comprising said plasmid or particles rescued therefrom, and uses thereof, especially in preventing or treating a tumor in a patient.

Abstract: Perfusion media are disclosed providing excellent cell density, titer and product quality for production of a therapeutic protein in a perfusion process.

Abstract: The invention relates to a recombinant measles virus expressing a heterologous amino acid sequence derived from an antigen of a determined RNA virus, said recombinant measles virus being capable of eliciting a humoral and/or cellular immune response against measles virus or against said RNA virus or against both measles virus and against said RNA virus. It also relates to the use of said recombinant measles virus for the preparation of immunogenic composition.

Abstract: A pharmaceutical composition which has a plurality of lipid nanoparticles that has a mean particle size of between 80 nm and 160 nm and contains a modified mRNA encoding a polypeptide. The lipid nanoparticles include a cationic lipid, a neutral lipid, a cholesterol, and a PEG lipid. The mRNA contains a 5?-cap, 5?-UTR, N1-methyl-pseudouridine, a 3?-UTR, and a poly-A region with at least 100 nucleotides.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: The subject invention pertains to methods and apparatus for the production and purification of cell products, such as immunoglobulins. One aspect of the invention is an integrated cell culture and purification apparatus for the growth and maintenance of cells and the harvest and purification of cell products, such as immunoglobulins. Thus, the apparatus integrates a cell culture function with a purification function. Other aspects of the invention pertain to an automated method for producing immunogenic compositions such as vaccines.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: A peptide nucleic acid (PNA) referred to herein as Sweet-P, compositions comprising the methods of making the same, mid methods of using the same, are described.

Abstract: Provided herein are methods of culturing an adherent mammalian cell in a shake tube using a plurality of microcarriers and batch re-feed perfusion, and various methods that utilize these culturing methods.

Abstract: The present disclosure describes compositions including polynucleotides encoding polypeptides which have been chemically modified by replacing the uridines with 5-methoxy-uridines to improve the compositions, engagement with translational machinery, mRNA half-life, translation efficiency, immune evasion, protein production capacity, secretion efficiency, accessibility to circulation, protein half-life and/or modulation of a cell's status, protein half-life and/or modulation of a cell's status function, and/or activity.

Abstract: The present disclosure provides compositions and methods useful for tissue engineering including a composition having chemically modified RNA (cmRNA) encapsulated in or complexed with a non-viral delivery vehicle and a biocompatible, bioresorbable scaffold and methods of using the composition to regenerate, for example, bone tissue.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: The invention relates to compositions and methods for the preparation, manufacture and therapeutic use of polynucleotides, primary transcripts and mmRNA molecules.

Abstract: The present invention provides a cell capable of high-yield production of polypeptides and a method for producing the same. The present invention relates to a method for producing a cell capable of high-yield production of a desired polypeptide, wherein a strongly taurine transporter-expressing cell into which DNA encoding the desired polypeptide has been introduced is cultured in the presence of a high concentration of methotrexate and a cell capable of high-yield production of the desired polypeptide is selected from among surviving cells.

Abstract: The invention relates to a cDNA molecule which encodes the nucleotide sequence of the full length antigenomic (+)RNA strand of a measles virus (MV) originating from an approved vaccine strain. It also concerns the preparation of immunogenic compositions using said cDNA.

Abstract: The present invention provides a method capable of producing a natural or recombinant protein in high yield. The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses alanine aminotransferase and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide.

Abstract: The present invention provides a method capable of producing a natural or recombinant protein at low cost. The present invention relates to a method of producing a polypeptide, comprising culturing a cell which strongly expresses a taurine transporter and has a transferred DNA encoding a desired polypeptide and thereby allowing the cell to produce the polypeptide. Hamster taurine transporter, a DNA encoding the same, a recombinant vector and a transformed cell are also provided.

Abstract: This invention relates to a method for producing a protein of interest, comprising introducing a protein expression vector which comprises a gene fragment a gene fragment comprising a DNA encoding a protein of interest and a selectable marker gene and transposon sequences at both terminals of the gene fragment, into a suspension mammalian cell; integrating the gene fragment inserted between a pair of the transposon sequences, into a chromosome of the mammalian cell to obtain a mammalian cell capable of expressing the protein of interest; and suspension-culturing the mammalian cell; and a suspension mammalian cell capable of expressing the protein of interest.

Abstract: The subject invention pertains to methods and apparatus for the production and purification of cell products, such as immunoglobulins. One aspect of the invention is an integrated cell culture and purification apparatus for the growth and maintenance of cells and the harvest and purification of cell products, such as immunoglobulins. Thus, the apparatus integrates a cell culture function with a purification function. Other aspects of the invention pertain to an automated method for producing immunogenic compositions such as vaccines.

Abstract: The instant invention relates to modulated lysine variant species compositions comprising a protein, e.g., an antibody, or antigen-binding portion thereof, and methods, e.g., cell culture and/or protein purification methods, for producing such modulated lysine variant species compositions. Methods for using such compositions to treat a disorder, e.g., a disorder in which TNF? is detrimental, are also provided.

Abstract: The present invention provides biomatrix scaffolds, a tissue extract enriched for extracellular matrix components and bound growth factors, cytokines and hormones, and methods of making and using same.

Abstract: The present invention relates to cells capable of expressing IDO, nucleic acid constructs for expression of IDO, cells comprising said constructs and methods of utilizing said cells in the treatment of diseases. In particular the present invention relates to cells which express IDO in the absence of exposure to IFN-gamma, and to their use in preparation and/or generation of immunomodulatory cells specific for an antigen.

Abstract: An improved system for large scale production of polypeptides in cell culture is provided. In accordance with the present invention, cells expressing a polypeptide of interest are grown in media that contain copper, glutamate or both. The use of such a system allows production of polypeptides in which misfolding and/or aggregation are reduced, and in which total glycosylation is increased. Polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical, agricultural or other commercial compositions.

Abstract: A method of inducing a specific immune response in a mammal, comprising: providing a first composition comprising isolated ubiquitinylated proteins in solution in the absence of membrane bound organelles, the isolated ubiquitinylated proteins comprising one or more specific antigens, and further comprising a threshold quantity of polyubiquitinylated short-lived proteins and polyubiquitinylated defective ribosomal products. The isolated ubiquitinylated proteins are affinity-purified from tumor-derived cells grown in culture, the tumor-derived cells being inhibited from degrading ubiquitinylated proteins via the proteasome while being grown in culture. In this way, highly immunogenic short-lived proteins and defective ribosomal products may be loaded onto dendritic cells for cross-presentation and priming of antigen-specific T cells restricted by either classical or non-classical MHC.

Abstract: The invention provides a process for generating an avian cell line of duck origin by a process comprising: —cultivating embryonic avian cells in cell culture medium, preferably comprising fetal calf serum (FBS) for more than 40 passages while reducing the concentration of FBS to 1-2% vol/vol FBS, —transferring passaged cells into cell culture medium having 0% FBS, —resulting in the generation of a non-embryonic avian cell line suitable for non-adherent growth, i.e. for growth in suspended culture.

Abstract: The present invention pertains to methods of preventing and eliminating trisulfide bonds in proteins such as antibodies. In one embodiment, trisulfide bonds in proteins are converted to disulfide bonds as part of chromatographic purification procedures. In another embodiment, the formation of trisulfide bonds in proteins is inhibited by implementation of methods described herein during the cell culture production of such proteins. In another embodiment, monoclonal antibodies are produced by the methods described herein.

Abstract: The invention relates to a cDNA molecule which encodes the nucleotide sequence of the full length antigenomic (+)RNA strand of a measles virus (MV) originating from an approved vaccine strain. It also contains the preparation of immunogenic compositions using said cDNA.

Abstract: A cell culture medium with high content of choline chloride is provided. The cell culture media further comprise only moderate amounts of amino acids, in particular the amount of glutamine in the cell culture media is limited. The cell culture media can be used for large scale production of polypeptides using cell cultures. The cell culture media with high content of choline chloride are particularly suitable for fed-batch cell culture whereby cell viabilities stay at a higher level for a longer time and high polypeptide titers although limited amounts of amino acids are used.

Abstract: Provided herein are methods for stable integration and/or expression of one or more recombinant polynucleotides in a host cell. The recombinant polynucleotides are typically integrated into the host genome at some native chromosomal integration sites. The integration can be mediated by homologous recombination or by using a hybrid recombinase targeting the specific chromosomal locations. The native chromosomal integration sites in the host cells, which support stable integration and strong transcription activities of foreign genes, are present within or adjacent to specific genes in the CHO genome, the ankyrin 2 gene (Ank2), cleavage and polydenylation specific factor 4 gene (Cpsf4), C-Mos gene, and Nephrocystin-1/Mal gene. Also provided are methods and nucleic acid molecules for inserting site-specific recombination sequences (chromosomal landing pads) into these specific chromosomal locations, engineered host cells containing chromosomal landing pads, methods and compositions (e.g., kits) therefore.

Abstract: Methods for producing a protein extract from cells, such as cells containing viral proteins, are provided. In general terms, the methods involve: increasing the pH of the cells to a pH of at least about pH 10.0 to produce an intermediate composition, and then, in the presence of a non-ionic detergent, neutralizing the pH of the intermediate composition to produce the protein extract. Kits and compositions for practicing the subject methods are also provided.

Abstract: Antagonists of human proprotein convertase subtilisin-kexin type 9 (“PCSK9”) are disclosed. The disclosed antagonists are effective in the inhibition of PCSK9 function and, accordingly, present desirable antagonists for use in the treatment of conditions associated with PCSK9 activity. The present invention also discloses nucleic acid encoding said antagonists, vectors, host cells, and compositions comprising the antagonists. Methods of making PCSK9-specific antagonists as well as methods of using the antagonists for inhibiting or antagonizing PCSK9 function are also disclosed and form important additional aspects of the present disclosure.

Abstract: The present invention relates to a process for producing a desired polypeptide using rat cells. Specifically, the present invention relates to a process for producing the polypeptide which comprises culturing rat cells such as YB2/3HL.P2.G11.16Ag.20 (hereinafter referred to as YB2/0), preferably rat cells to which a recombinant DNA comprising DNA encoding a desired polypeptide such as an immunologically functional molecule is introduced, in a medium which does not contain serum (hereinafter referred to as a serum-free medium). Among the desired polypeptides obtained by the process of the present invention, an antibody obtained by using a transformant of YB2/0 has a high antibody-dependent cell-mediated cytotoxic activity (hereinafter sometimes referred to as ADCC activity) and is useful as a pharmaceutical agent.

Abstract: The present invention relates to skin care compositions, including cosmeceuticals, for topical application, and more particularly, a skin cream, comprising exosomes and cell culture medium conditioned by cells grown in two-dimensional culture. Also included are methods of making and using such compositions and kits comprising the skin cream therein.

Abstract: The present invention provides an Avian adeno-associated virus (AAAV) virus and vectors and particles derived therefrom. In addition, the present invention provides methods of delivering a nucleic acid to a cell using the AAAV vectors and particles. Methods of isolating the AAAV are provided.

Abstract: The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.

Abstract: The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.

Abstract: A protein transduction method for efficiently delivery of exogenous proteins into mammalian cells is invented, which has the capability of targeting different cellular compartments and protection from degradation of the delivered proteins from cellular proteases. A composition for treat proteins has cation reagents, lipids and enhancers in a carrier. The method can be used in a number of ways including: production of large quantities of properly folded, post-translationally modified proteins using mammalian cell machinery, a in-cell fluorescence spectroscopy and imaging using small molecule fluorophores and a in-cell NMR spectroscopy using living mammalian cells. The method permits cell biology at atomic resolution that is physiologically and pathological relevant and permits protein therapy to treat human diseases. The method can also be used to deliver exogenous protein inside mammalian cells, wherein the exogenous proteins follow a similar secretion pathway as that of the endogenous protein.

Abstract: A use of a signal peptide for producing a recombinant polypeptide of interest in an expression system, the signal peptide includes at least 12 amino acids of formula (I): (X1)iX2X3X4SX5X6X7, wherein: X1 is a peptide containing from 3 to 6 amino acids, i equal to 0 or 1, X2 is a peptide containing from 3 to 9 hydrophobic amino acids, X3 is a peptide containing from 3 to 5 amino acids, the peptide including at least 3 contiguous or non-contiguous leucines X4 is a peptide containing from 2 to 5 amino acids chosen from Ala, Thr, Ser, Gln, Ile, Met, X5 is Ala or Val, X6 is Gln, Asn or His, X7 is Ala or Cys, provided that when the signal peptide originates from a natural precursor of a specific protein, the polypeptide of interest is different from the protein.

Abstract: The present invention relates to an eukaryotic expression vector comprising a nucleotide sequence encoding for the human long pentraxin PTX3 protein under the control of an effective promoter and a nucleotide sequence encoding for a selectable marker, recombinant human cell able to provide expression of proteins encoded by the vector and method for the production of the human long pentraxin PTX3 protein.

Abstract: A bioscaffold made from an isolated cardiac fibroblast-derived 3-dimensional extracellular matrix (ECM) is disclosed. The bioscaffold can be used as an epicardial patch for the delivery of therapeutic cells into myocardial tissue. Methods of making the 3-dimensional extracellular matrix using cultured cardiac fibroblasts are also disclosed.

Abstract: The present invention provides, among other things methods of increasing cell density, viability and/or titer in a cell culture including steps of adding a composition comprising iron to the cell culture.

Abstract: The present invention relates to methods of producing a Factor VIII polypeptide in mammalian cell cultures.

Abstract: A method of mRNA production for use in transfection is provided, that involves in vitro transcription of PCR generated templates. This RNA can efficiently transfect different kinds of cells. This approach results in increased efficiency (fidelity and productivity) of mRNA synthesis and is less time consuming because it does not require cloning, and also consequently eliminates the unwanted errors and effects related to RNA made on DNA templates obtained with cloning techniques. The results of transfection of RNAs demonstrate that RNA transfection can be very effective in cells that are exceedingly difficult to transfect efficiently with DNA constructs. The method can be used to deliver genes into cells not- or only poorly transfectable for DNA, in vitro and in vivo.

Abstract: An assembly capable of capturing and purifying expressed biological products during or at the end of a bioreaction cycle is disclosed wherein a binding resin is kept separated from the contents of the bioreactor allowing capturing, harvesting and purification of biological products in a bioreactor; the invention additionally provides means of removing undesirable metabolic products as well as provides for efficient loading of chromatography columns.

Abstract: The present invention pertains to methods of preventing and eliminating trisulfide bonds in proteins such as antibodies. In one embodiment, trisulfide bonds in proteins are converted to disulfide bonds as part of chromatographic purification procedures. In another embodiment, the formation of trisulfide bonds in proteins is inhibited by implementation of methods described herein during the cell culture production of such proteins. In another embodiment, monoclonal antibodies are produced by the methods described herein.

Abstract: Lentiviral vectors modified at the 5? LTR or both the 5? and 3? LTR's are useful in the production of recombinant lentivirus vectors. Such vectors can be produced in the absence of a functional tat gene. Multiple transformation of the host cell with the vector carrying the transgene enhances virus production.

Abstract: Disclosed is a method for introducing a gene into a target cell using a retrovirus vector, which is simple and highly efficient. Specifically disclosed is a method for introducing a gene into a target cell using a retrovirus vector, which comprises the steps of (a) placing a liquor containing a retrovirus vector having a foreign gene carried thereon into a culture vessel on which a retrovirus-binding substance has been immobilized, and incubating the liquor at a temperature lower than 25° C. for 4 hours or longer, thereby producing a culture vessel having the retrovirus vector bound thereto, and (b) adding a target cell to the culture vessel that has been produced in step (a) and incubating the culture vessel. The gene introduction method is simple and highly efficient, and is useful particularly in the fields of medicine, cell technology, gene technology, embryologic technology and the like.