Abstract: The present invention relates to polynucleotides comprising a first nucleic acid sequence for a chromatin element, which is capable of enhancing expression, and at least one second nucleic acid sequence comprising a curved origin motif. Furthermore, the invention relates to a host cell, a non-human transgenic organism, a vector and a kit comprising the aforementioned polynucleotide. Moreover, the invention relates to methods for expressing a polynucleotide of interest.

Abstract: Acellular compositions for treating inflammation, comprising two or more of IL1-ra, sTNF-R1, sTNF-RII, IGF-I, EGF, HGF, PDGF-AB, PDGF-BB, VEGF, TGF-?1, and sIL-1RII. Components of the acellular compositions may be derived from biologic materials, such as blood clots and urine. Components may also be obtained from cell cultures.

Abstract: The present invention relates to a medium for the cultivation of eukaryotic cells, the medium comprising as (an) additive(s) DMSO, N-acetylmannosamine (NAcMan), N-acetylglucosamine (NAcGlc), or any combination of two or more of these additives, including the combination of NAcMan and NAcGlc.

Abstract: The purpose of the present invention is to develop a method for producing a large amount of an insoluble aggregate that is equivalent to an insoluble aggregate formed in the brain of a patient. A method of producing an insoluble aggregate of a neurodegenerative-disease-related protein according to the present invention comprises the steps of: (1) introducing an insoluble fraction originated from the brain of a neurodegenerative disease patient into a cultured cell in which the neurodegenerative-disease-related protein can be expressed in a constitutive manner; (2) culturing the cultured cell into which the insoluble fraction has been introduced; and (3) extracting separating an insoluble fraction from the cultured cell. Optionally, the method may additionally comprise a step of amplifying the insoluble aggregate of the neurodegenerative-disease-related protein in the cultured cell.

Abstract: The present invention relates to methods and systems for optimization of dilution of a viscous starting material to isolate and/or concentrate the product of interest from the starting source material such that the process minimizes the volume of diluent and the total volume of the waste stream generated during the process as well as maximizing the yield of desired product. The system employs cross-flow filtration modules with sub-channels that are equidistant to the inlet and outlet of said modules and such modules are characterized by optimal channel height, optimal transmembrane pressure, etc., which are selected in order to achieve the best combination of product quality and production yield.

Abstract: Differentiated cholinergic cells having motor neuron-like morphology and increased sensitivity to botulinum neurotoxin are provided herein. Methods of using such differentiated cells for detecting neurotoxin are also provided.

Abstract: A method is disclosed for releasing the transcriptional regulation caused by a repeated sequence in a gene, a kit therefor and so on to thereby establish a system capable of producing a protein in a large amount.

Abstract: This invention relates to a cell culture process for the production of polypeptides in mammalian CHO cells characterized by one or more temperature and pH shifts which are adjusted in respect to their timing and step size to reduce cell death, increase product yield and improve product quality.

Abstract: [Summary] The present invention relates to a protein production accelerating agent that has enabled to largely increase the produced amount of a desired protein by adding polysaccharides to a medium for animal cells containing a serum or serum alternative, and a production method of a protein using a medium containing the protein production accelerating agent.

Abstract: Complexes that contain RSV F ectodomain polypeptides and methods for making the complexes are disclosed. The RSV F ectodomain polypeptides can be in the prefusion form.

Abstract: A serum-free C3A clonal cell line and methods for generating the same are provided. The C3A cell line has a reduced doubling time in serum-free medium compared to a corresponding C3A cell line from which it is derived. Methods using the cells of the serum-free C3A clonal cell line for the production, expression and recovery of harvestable polypeptides, screening compounds for metabolic activity, studying enteric disease and for use in a bio-artificial liver device are also provided.

Abstract: The present invention provides methods of improving the levels and stability of expression of interleukin-12 family cytokine polypeptides by expressing the alpha and beta subunits of the polypeptides at their determined relative molar ratios that increase the levels and stability of expression of the heterodimer, e.g., in comparison to heterodimer expressed at an equimolar ratio.

Abstract: The present invention describes methods and processes for the production of proteins by animal cell or mammalian cell culture. In one aspect, the methods comprise the growth of cells in a growth factor/protein/peptide free medium. In another aspect, the methods comprise the addition of growth factors during the production phase. The methods sustain a high viability of the cultured cells, and can yield an increased end titer of protein product, and a high quality of protein product.

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: Provided is an antibody production method whereby it is possible to repeatedly acquire antibodies produced by fish without killing the fish. Specifically provided is an antibody production method whereby it is possible to repeatedly acquire the antibodies produced by a fish, without killing the fish, by administering antigens to fish that have blisters.

Abstract: In accordance with the present invention, there are provided fully human monoclonal antibodies against human cytotoxic T-lymphocyte antigen 4 (CTLA-4). Nucleotide sequences encoding and amino acid sequences comprising heavy and light chain immunoglobulin molecules, particularly contiguous heavy and light chain sequences spanning the complementarity determining regions (CDRs), specifically from within FR1 and/or CDR1 through CDR3 and/or within FR4, are provided. Further provided are antibodies having similar binding properties and antibodies (or other antagonists) having similar functionality as antibodies disclosed herein.

Abstract: The invention relates to a process for the culturing of cells by continuous perfusion culturing of a cell culture comprising cell culture medium and cells, wherein cell culture medium is added to the cell culture, the cell culture is circulated over a filter module comprising hollow fibers resulting in an outflow of liquid having a lower cell density than the cell culture and the flow within the filter module is an alternating tangential flow. Preferably, culture medium is added at a particular perfusion rate and/or biomass is removed form the culture at least once. The method is especially suitable for the culturing of aggregating cells. The invention also relates to such a process wherein a biological substance, preferably an antibody, is produced by the cells, which biological substance may be further purified in downstream processing.

Abstract: The present invention relates to a method of controlling quality and quantity of posttranslational modification of a recombinantly produced polypeptide/protein (glycoprotein), wherein the posttranslational modification affects the glycosylation profile and/or the acidic variants profile, as manifested in CEX profiles, wherein the polypeptide/protein (glycoprotein) production is in eukaryotic host cells, the method comprising the following steps: a) cultivating the eukaryotic cells in a suitable medium under conditions which allow the expression of the polypeptide/protein, wherein the content of the dissolved CO2 (pCO2) in the medium is at a first value during the initial growth phase of the eukaryotic cells, allowing the eukaryotic cells to grow, and b) increasing or decreasing the content of the dissolved CO2 (pCO2) in the medium during the production phase of the eukaryotic cells to a second value.

Abstract: The present invention relates to methods of host cell transduction utilizing ecotropic retroviral vector particles. The retroviral vector particle may comprise an envelope of Friend murine leukaemia virus, in particular the envelope encoded by molecular clone PVC-211 and the host cell may be engineered to recombinantly express the Reel receptor. The retroviral vector particles and methods of the invention can be used to introduce expressible polynucleotide sequences of interest into host cells with high efficiency. This results in protein production methods with higher yield (mg/L) and a reduction in manufacturing costs that could be used in a range of applications including for example, the production of therapeutic proteins, vaccines and antibodies.

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: This invention provides a TNFR2 expression-inducing composition including as an active ingredient a peptide having TNFR2 expression-inducing activity, and a method for producing cells that express TNFR2 selectively by use of the composition. The cell production method provided by this invention includes: culturing at least one species of cells capable of expressing TNF receptor 2, and supplying the cells with a synthetic peptide consisting of a nuclear localization signal sequence (NLS) or a nucleolar localization signal sequence (NoLS) to enhance TNFR2 expression in the cells.

Abstract: The present disclosure relates to methods of decreasing lactate production in cell culture using divalent transitional metallic salts. The present disclosure also relates to a method of producing polypeptide by adding divalent transitional metallic salt to the cell culture medium for reducing lactate accumulation followed by fermenting and recovering the polypeptide.

Abstract: The invention relates to an immunogenic composition comprising a recombinant vector characterized in that it comprises a polynucleotide comprising the cis-acting central initiation region (cPPT) and the cis-acting termination region (CTS), these regions being of retroviral or retroviral-like origin, said vector comprising in addition a defined nucleotide sequence (transgene or sequence of interest) and regulatory signals of retrotranscription, expression and encapsidation of retroviral or retroviral-like origin, wherein the composition is capable of inducing or of stimulating a cell-mediated response for instance a CTL (Cytotoxic T Lymphocytes) response or a CD4 response, against one or several epitopes encoded by the transgene sequence present in the vector.

Abstract: The invention relates to a bioreactor (1) for cell culture on a three-dimensional substrate, comprising a culture chamber (2), the inner walls of which form a vertical duct, preferably, tapered, with a diameter that widens regularly form the duct inlet to the duct outlet, means (3, 4) enabling the culture medium to flow in said vertical duct. The invention also relates to the advantageous use of these bioreactors in tissue engineering, for the production of tissue grafts, notably a bone or cartilage graft.

Abstract: The present invention relates to methods for the production of heat shock protein complexes for use in vaccine compositions. In particular, there is provided a method for increasing the level and immunogenicity of heat shock protein complexes produced in cells by subjecting the cells to specific stress inducing stimuli. The invention further extends to the use of heat shock protein complexes produced according to the methods of the invention in the preparation of vaccine compositions for the prevention and treatment of infectious diseases and cancerous conditions.

Abstract: Various embodiments of the invention provide methods of treating cancer. Many embodiments provide methods of treating cancer using stem cells. In one embodiment the method comprises removing cancer cells from a patient and culturing the cancer cells in the presence of stem cells under conditions such that the stem cells differentiate into stem cell-derived cancer cells (SCDCC) which can comprise stem cell derived non-invasive and/or invasive cancer cells. An amount of the SCDCC is introduced into the patient sufficient to induce the patient's immune system to produce antibodies which inhibit or destroy the patient's cancer cells. The SCDCC may also be genetically modified to produce various immune stimulating proteins which enhance the patient's immune response to the cancer cells and improves the efficacy of the SCDNIC in treating the patient's cancer.

Abstract: A method of enhancing the formation of extracellular matrix in culture. Cells in culture secrete most of the collagen into the media as unprocessed procollagen, i.e., the cells do not convert procollagen to collagen. In contrast, normal extracellular matrix deposition involves procollagen processing to collagen, fibril assembly and deposition into the cell layer to form a collagenous extracellular matrix. The addition of certain growth factors and the addition of a thin layer of a certain volume exclusion agent on top of the cells dramatically enhances the conversion of procollagen to collagen and will increase the amount of collagen and extracellular matrix associated with the cells. This invention advances bioengineering of connective tissues for medical applications that require an extensive and functional extracellular matrix with high tensile strength such as those in the cornea stroma, skin, tendons, ligaments, articular cartilage and the intervertebral disks.

Abstract: The invention provides compositions and methods for producing an immunogenic agent from a host cell. In various embodiments, the immunogenic agent is a polypeptide, an antigen, a virus particle, or a vaccine In one aspect, the invention provides for a method for producing an immunogenic agent from a host cell. The method generally comprises contacting the cell with a RNA effector molecule, a portion of which is complementary to a target gene, maintaining the cell in a large-scale bioreactor for a time sufficient to modulate expression of the target gene, wherein the modulation enhances production of the immunogenic agent from the cell, and isolating the immunogenic agent from the cell.

Abstract: A glycosyl transferase from Chinese hamster and related methods are described.

Abstract: The disclosure provides methods and compositions useful for culturing stem cell including embryonic stem cells, adult stem cells, and embryonic germ cells.

Abstract: Mice, embryos, cells, and tissues having a restricted immunoglobulin heavy chain locus and an ectopic sequence encoding one or more ADAM6 proteins are provided. In various embodiments, mice are described that have humanized endogenous immunoglobulin heavy chain loci and are capable of expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof that is functional in a male mouse. Mice, embryos, cells, and tissues having an immunoglobulin heavy chain locus characterized by a single human VH gene segment, a plurality of human DH gene segments and a plurality of human JH gene segments and capable expressing an ADAM6 protein or ortholog or homolog or functional fragment thereof are also provided.

Abstract: The invention relates generally to activatable antibodies that include a masking moiety (MM), a cleavable moiety (CM), and an antibody (AB) that specifically binds to epidermal growth factor receptor (EGFR), and to methods of making and using these anti-EGFR activatable antibodies in a variety of therapeutic, diagnostic and prophylactic indications.

Abstract: A method for fabricating a micro-organ device comprises providing a microscale support having one or more microfluidic channels and one or more micro-chambers for housing a micro-organ and printing a micro-organ on the microscale support using a cell suspension in a syringe controlled by a computer-aided tissue engineering system, wherein the cell suspension comprises cells suspended in a solution containing a material that functions as a three-dimensional scaffold. The printing is performed with the computer-aided tissue engineering system according to a particular pattern. The micro-organ device comprises at least one micro-chamber each housing a micro-organ; and at least one microfluidic channel connected to the micro-chamber, wherein the micro-organ comprises cells arranged in a configuration that includes microscale spacing between portions of the cells to facilitate diffusion exchange between the cells and a medium supplied from the at least one microfluidic channel.

Abstract: While a desired protein is prepared by culturing an animal cell that produces the protein to cause the protein to be produced, level of heterogeneity components of the protein is modulated by performing the culture at a normal culture temperature for a certain period and then continuing the culture at a culture temperature lowered to 25-35° C.

Abstract: The present invention discloses isolated nucleic acid molecules encoding a hyperimmune serum reactive antigen or a fragment thereof as well as hyperimmune serum reactive antigens or fragments thereof from E. faecalis, methods for isolating such antigens and specific uses thereof.

Abstract: The present invention relates to the field of cell culture technology and relates to methods of replicating/cloning cells, preferably cell lines which are important for the production of biopharmaceuticals. The invention also relates to methods of preparing proteins using cells that have been obtained and replicated by single cell deposition and compositions which make it possible to replicate individual cells. By using IGF particularly in conjunction with HSA in the culture medium after recloning, the recloning efficiency and hence the quantity of clones obtained can be increased significantly.

Abstract: The invention relates to a process for the culturing of cells by continuous perfusion culturing of a cell culture comprising cell culture medium and cells, wherein cell culture medium is added to the cell culture, the cell culture is circulated over a filter module comprising hollow fibers resulting in an outflow of liquid having a lower cell density than the cell culture and the flow within the filter module is an alternating tangential flow. Preferably, culture medium is added at a particular perfusion rate and/or biomass is removed form the culture at least once. The method is especially suitable for the culturing of aggregating cells. The invention also relates to such a process wherein a biological substance, preferably an antibody, is produced by the cells, which biological substance may be further purified in downstream processing.

Abstract: The present invention relates to a process for the production of a haemostasis protein by continuous perfusion culturing of a cell culture in suspension, said cell culture expressing said haemostasis protein into said culture suspension, wherein the cell culture flows across a filter module, which filter module leads to a harvest port, the filter module having a mesh size of from 0.1 to 2.9 ?m allowing passage across of the haemostasis protein and wherein the flow across the filter module is an alternating tangential flow. The invention also relates to a protein produced by the process of the invention.

Abstract: The present invention is directed to a method of producing compositions including embryonic proteins. The method includes culturing cells under hypoxic conditions on a biocompatible three-dimensional surface in vitro. The culturing method produces both soluble and non-soluble fractions, which may be used separately or in combination to obtain physiologically acceptable compositions useful in a variety of medical and therapeutic applications.

Abstract: The present invention provides a novel expression vector which comprises a gene of interest, a nuclear anchoring element, and at least one inverted repeat element, preferably two inverted repeat elements. The expression vector is an episomal vector capable of transfecting a mammalian cell. The present invention further provides a method for enhancing gene expression by transfecting the expression vector to a mammalian cell, preferably a human cell.

Abstract: The present invention is directed to a method of producing compositions including embryonic proteins. The method includes culturing cells under hypoxic conditions on a biocompatible three-dimensional surface in vitro. The culturing method produces both soluble and non-soluble fractions, which may be used separately or in combination to obtain physiologically acceptable compositions useful in a variety of medical and therapeutic applications.

Abstract: Disclosed are methods and compositions for early diagnosis, monitoring and treatment of retinal dystrophy, age-related macular degeneration, Bardet-Biedel syndrome, Bassen-kornzweig syndrome, best disease, choroidema, gyrate atrophy, congenital amourosis, refsun syndrome, stargardt disease and Usher syndrome. In particular, the invention relates to a protein, termed “Rdcvf1,” that is differentially transcribed and expressed in subjects suffering from retinal dystrophies and the like, such as retinal dystrophy and age-related macular degeneration compared with nonsufferers, antibodies which recognize this protein, and methods for diagnosing such conditions.

Abstract: Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder. Additionally, the medium is formulated with a pharmaceutically acceptable carrier as a vehicle for internal administration, applied directly to a food item or product, or formulated with a salve or ointment for topical applications. Also, the medium may be further processed to concentrate or reduce one or more factors or components contained within the medium.

Abstract: The present invention relates to the production of recombinant glycoproteins or antibodies that have an improved glycosylation profile and effector functions such as ADCC and/or CDC. The present invention is in particular related to the production of glycoproteins or antibodies having a valuable glycosylation profile, especially a low fucose level and/or a high oligomannose level and/or presence of sialic acid to the glycans.

Abstract: The present invention relates to a pharmaceutical composition for the prevention or treatment of immune disorders and inflammatory diseases, comprising stem cells that are generated by culturing stem cells expression Nucleotide-binding Oligomerization Domain protein 2 (NOD2) with a NOD2 agonist or a culture thereof.

Abstract: Novel products comprising conditioned cell culture medium compositions and methods of use are described. The conditioned cell medium compositions of the invention may be comprised of any known defined or undefined medium and may be conditioned using any eukaryotic cell type. Once the cell medium of the invention is conditioned, it may be used in any state. Physical embodiments of the conditioned medium include, but are not limited to, liquid or solid, frozen, lyophilized or dried into a powder. Additionally, the medium is formulated with a pharmaceutically acceptable carrier as a vehicle for internal administration, applied directly to a food item or product, or formulated with a salve or ointment for topical applications. Also, the medium may be further processed to concentrate or reduce one or more factors or components contained within the medium.

Abstract: An in vitro process for the preparation of antibodies of the IgG type, comprising the steps of: i. challenging dendritic cells obtained from a donor with an antigen against which the antibodies to be prepared are directed; ii. challenging CD4+ T cells obtained from the same donor with fragments of the antigen presented by the dendritic cells from step L, with the proviso that the dendritic cells are ones essentially not secreting interferon-? (IFN-?) and interleukin 12 (IL-12), to generate antigen-specific CD4+ Th2 cells; iii. challenging with the antigen a B cell population obtained from the same donor and including a sufficient proportion of naive B cells; and iv. contacting the antigen-specific CD4+ Th2 cells according to ii. with at least a fraction of the B cell population from iii. to form an antigen-specific plasma cell; v. immortalizing said antigen-specific plasma cell; and vi. isolating the IgG antibodies formed by said antigen-specific plasma cell.

Abstract: Described are means and methods for removing a proteolytic cleavage site from a protein, the method comprising providing a cell that expresses pre-mRNA encoding the protein with an anti-sense oligonucleotide that induces skipping of the exonic sequence that encodes the proteolytic cleavage site, and allowing translation of mRNA produced from the pre-mRNA.

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 provides an in vitro method to predict the effect of an exogenous element on an animal or human organism. This method is based on the analysis of variations of an organelle behaviour represented by at least two characteristics in isolated cells induced by the contact with the exogenous element and the determination of the cluster membership of the exogenous element which is representative of the effect of said element. Analyzed characteristics are selected from the group consisting of the motility, the morphology, the relationship with the cell cytoskeleton and the membrane permeability of said organelle. This method may be used to predict the efficacy or the toxicity of an element, such as a drug.