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Titlebook: Heterologous Protein Production in CHO Cells; Methods and Protocol Paula Meleady Book 2017 Springer Science+Business Media LLC 2017 CRISPR.

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Cloning of Single-Chain Antibody Variants by Overlap-Extension PCR for Evaluation of Antibody Expretions and assembled by a single overlap-extension PCR reaction. The amplified product is then cloned into a mammalian expression vector suitable for high-titer transient gene expression. This workflow can be applied to any antibody sequence by adapting the specific primer sequences to the antibody of choice.
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Improved CHO Cell Line Stability and Recombinant Protein Expression During Long-Term Culture,e stability of the recombinant CHO cells over long-term culture. These procedures include the following; western blotting, ELISA to evaluate protein production, real-time PCR to analyze plasmid and mRNA copy numbers, and fluorescent in situ hybridization (FISH) to assess the location of the inserted plasmid on host cell chromosomes.
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Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension,incubation at 31 °C with agitation by orbital shaking. We also describe an alternative method in which 90% of the pDNA is replaced by nonspecific (filler) DNA, and the production phase is performed at 31 °C in the presence of 0.25% N, N-dimethylacetamide (DMA).
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Selection of High-Producing Clones Using FACS for CHO Cell Line Development,powerful, high-throughput technique that facilitates multiparametric characterization and isolation of individual cell clones from heterogeneous populations. Here, we describe a FACS-based method for section of high-producing CHO cell clones.
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Characterization of Host Cell Proteins (HCPs) in CHO Cell Bioprocesses, while estimation of HCP clearance/presence can be achieved by comparing 2D-PAGE images of samples and by undertaking western blotting of 2D-PAGE analyzed samples. Here, we describe the analyses of HCP content using these methodologies.
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