A5000736115- Viral Infectious Diseases and Gene Expression in Insects
- Monoclonal and Polyclonal Antibodies Research
- Virus-based gene therapy research
- Protein purification and stability
- CRISPR and Genetic Engineering
- RNA and protein synthesis mechanisms
- Viral Infections and Immunology Research
- Molecular Biology Techniques and Applications
- Transgenic Plants and Applications
- Bacteriophages and microbial interactions
- RNA Interference and Gene Delivery
- RNA Research and Splicing
- CAR-T cell therapy research
- Gene Regulatory Network Analysis
AstraZeneca (United Kingdom)
2020-2023
Granta Design (United Kingdom)
2017
Comprehensive de novo-design of complex mammalian promoters is restricted by unpredictable combinatorial interactions between constituent transcription factor regulatory elements (TFREs). In this study, we show that modular binding sites do not function cooperatively can be identified analyzing host cell expression profiles, and subsequently testing cognate TFRE activities in varying homotypic heterotypic promoter architectures. TFREs displayed position-insensitive, additive within a...
Real-time quantitative PCR (qPCR) is the standard method for determination of relative changes in mRNA transcript abundance. Analytical accuracy, precision and reliability are critically dependent on selection internal control reference genes. In this study, authors have identified optimal genes that can be utilised universally qPCR analysis CHO cell mRNAs. Initially, transcriptomic datasets were analysed to identify eight endogenous exhibited high expression stability across four distinct...
To successfully engineer mammalian cells for a desired purpose, multiple recombinant genes are required to be coexpressed at specific and optimal ratio. In this study, we hypothesized that synthetic promoters varying in transcriptional activity could used create single multigene expression vectors coexpressing predictable relative stoichiometry. A library of 27 constructs was created comprising three discrete fluorescent reporter gene units fixed series, each under the control either...
ABSTRACT Monoclonal antibodies (mAbs) contain short N‐terminal signal peptides on each individual polypeptide that comprises the mature antibody, targeting them for export from cell in which they are produced. The peptide is cleaved heavy chain (Hc) and light (Lc) after translocation to ER prior secretion. This process generally highly efficient, producing a high proportion of correctly Hc Lc polypeptides. However, mis‐cleavage can occur, resulting truncation or elongation at N‐terminus Lc....
The output from protein biomanufacturing systems is a function of total host cell biomass synthetic capacity and recombinant production per unit biomass. In this study, we describe how these two properties can be simultaneously optimized via design product-specific combination DNA parts to maximize flux through the pathway use chassis with an increased capability synthesize both product Using secreted alkaline phosphatase (SEAP) in Chinese hamster ovary cells as our example, demonstrate...
The manufacture of bispecific antibodies by Chinese hamster ovary (CHO) cells is often hindered lower product yields compared to monoclonal antibodies. Recently, reactive oxygen species have been shown negatively impact antibody production. By contrast, strategies boost cellular antioxidant capacity appear be beneficial for recombinant protein expression. With this in mind, we generated a novel hydrogen peroxide evolved host using directed cell evolution. Here demonstrate that has heritable...
Aggregation of therapeutic bispecific antibodies negatively affects the yield, shelf-life, efficacy and safety these products. Pairs stable Chinese hamster ovary (CHO) cell lines produced two difficult-to-express with different levels aggregated product (10-75% aggregate) in a miniaturised bioreactor system. Here, transcriptome analysis was used to interpret biological causes for aggregation identify strategies improve yield quality. Differential expression- gene set revealed upregulated...
High levels of protein expression are key to the successful development and manufacture a therapeutic antibody. Here, we describe two related antibodies, Ab001 Ab008, where shows markedly lower level relative Ab008 when stably expressed in Chinese hamster ovary cells. We use single-gene vectors structural analysis show that reduced titer is associated with VL CDR2 Ab001. adopted approaches improve First, used mutagenesis change single amino-acid residues back equivalent but this resulted...
When expressing complex biotherapeutic proteins, traditional expression plasmids and methods may not always yield sufficient levels of high-quality product. High-strength viral promoters commonly used for recombinant protein (rProtein) production in mammalian cells allow maximal expression, but provide limited scope to alter their transcription dynamics. However, synthetic designed tunable transcriptional activity offer a plasmid engineering approach more precisely regulate product quality,...
Abstract Control of mammalian recombinant protein expression underpins the in vitro manufacture and vivo performance all biopharmaceutical products. However, routine optimization levels these applications is hampered by a paucity genetic elements that function predictably across varying molecular formats host cell contexts. Herein, we describe synthetic components are specifically built to simplify bioindustrial cassette design processes. Synthetic G-quadruplex with sequence feature...
Introns are included in genes encoding therapeutic proteins for their well-documented function of boosting expression. However, mis-splicing introns recombinant immunoglobulin (IgG) heavy chain (HC) transcripts can produce amino acid sequence product variants. These variants affect quality; therefore, purification process optimization may be needed to remove them, or if they cannot removed, then in-depth characterization must carried out understand effects on biological activity. In this...