In the field of therapeutic antibody production, diversification of fed-batch strategies is flourishing in response to the market demand. All manufacturing approaches tend to follow the generally accepted dogma of increasing titer since it directly increases manufacturing output. While titer is...
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Cleavage-Independent HIV-1 Trimers From CHO Cell Lines Elicit Robust Autologous Tier 2 Neutralizing Antibodies
Native flexibly linked (NFL) HIV-1 envelope glycoprotein (Env) trimers are cleavage-independent and display a native-like, well-folded conformation that preferentially displays broadly neutralizing determinants. The NFL platform simplifies large-scale production of Env by eliminating the need to...
Read MoreKeywords: recombinant protein expression HIV-1 immunogenicity viral fusion proteins adjuvants vaccines
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MAR-Mediated Transgene Integration into Permissive Chromatin and increased expression involve an SD-MMEJ-like DNA repair Pathway.
Untargeted plasmid integration into mammalian cell genomes remains a poorly understood and inefficient process. The formation of plasmid concatemers and their genomic integration has been ascribed either to non-homologous end-joining (NHEJ) or homologous recombination (HR) DNA repair pathways....
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Use of human MAR elements to improve retroviral vector production.
Retroviral vectors have many favorable properties for gene therapies, but their use remains limited by safety concerns and/or by relatively lower titers for some of the safer self-inactivating (SIN) derivatives. In this study, we evaluated whether increased production of SIN retroviral vectors can...
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Epigenetic regulatory elements: recent advances in understanding their mode of action and use for recombinant protein production in mammalian cells.
Successful generation of high producing cell lines requires the generation of cell clones expressing the recombinant protein at high levels and the characterization of the clones’ ability to maintain stable expression levels. The use of cis-acting epigenetic regulatory elements that improve this...
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Epigenetic regulatory elements: Recent advances in understanding their mode of action and use for recombinant protein production in mammalian cells.
Successful generation of high producing cell lines requires the generation of cell clones expressing the recombinant protein at high levels and the characterization of the clones’ ability to maintain stable expression levels. The use of cis-acting epigenetic regulatory elements that improve this...
Read MoreKeywords: Epigenetics Matrix attachment region Recombinant protein production Transgene silencing Ubiquitously-acting chromatin opening element
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CHO cell engineering to prevent polypeptide aggregation and improve therapeutic protein secretion.
The ability to efficiently produce recombinant proteins in a secreted form is highly desirable and cultured mammalian cells such as CHO cells have become the preferred host as they secrete proteins with human-like post-translational modifications. However, attempts to express high levels of...
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MAR elements and transposons for improved transgene integration and expression.
Reliable and long-term expression of transgenes remain significant challenges for gene therapy and biotechnology applications, especially when antibiotic selection procedures are not applicable. In this context, transposons represent attractive gene transfer vectors because of their ability to...
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Rapid Production of Functional Proteins of a Combinatorial IgG Library in CHO Cells
Recombinant DNA (rDNA) technologies provide a wide range of tools for producing a broad array of recombinant proteins. Since the early 1970s, the biotechnology industry has harnessed those tools — together with genetic engineering and genomics — for developing new classes of innovative and...
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High-level transgene expression by homologous recombination-mediated gene transfer.
Gene transfer and expression in eukaryotes is often limited by a number of stably maintained gene copies and by epigenetic silencing effects. Silencing may be limited by the use of epigenetic regulatory sequences such as matrix attachment regions (MAR). Here, we show that successive transfections...
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