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Heterologous Protein Production in CHO Cells

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Cover of 'Heterologous Protein Production in CHO Cells'

Table of Contents

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    Book Overview
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    Chapter 1 Strategies and Considerations for Improving Expression of “Difficult to Express” Proteins in CHO Cells
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    Chapter 2 Glycoengineering of CHO Cells to Improve Product Quality
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    Chapter 3 Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension
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    Chapter 4 Cloning of Single-Chain Antibody Variants by Overlap-Extension PCR for Evaluation of Antibody Expression in Transient Gene Expression
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    Chapter 5 Anti-Apoptosis Engineering for Improved Protein Production from CHO Cells
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    Chapter 6 Conditional Knockdown of Endogenous MicroRNAs in CHO Cells Using TET-ON-SanDI Sponge Vectors
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    Chapter 7 Application of CRISPR/Cas9 Genome Editing to Improve Recombinant Protein Production in CHO Cells
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    Chapter 8 Improved CHO Cell Line Stability and Recombinant Protein Expression During Long-Term Culture
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    Chapter 9 Selection of High-Producing Clones Using FACS for CHO Cell Line Development
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    Chapter 10 The ‘Omics Revolution in CHO Biology: Roadmap to Improved CHO Productivity
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    Chapter 11 A Bioinformatics Pipeline for the Identification of CHO Cell Differential Gene Expression from RNA-Seq Data
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    Chapter 12 Filter-Aided Sample Preparation (FASP) for Improved Proteome Analysis of Recombinant Chinese Hamster Ovary Cells
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    Chapter 13 Phosphopeptide Enrichment and LC-MS/MS Analysis to Study the Phosphoproteome of Recombinant Chinese Hamster Ovary Cells
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    Chapter 14 Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture
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    Chapter 15 Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells
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    Chapter 16 Characterization of Host Cell Proteins (HCPs) in CHO Cell Bioprocesses
Attention for Chapter 3: Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension
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Chapter title
Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension
Chapter number 3
Book title
Heterologous Protein Production in CHO Cells
Published in
Methods in molecular biology, January 2017
DOI 10.1007/978-1-4939-6972-2_3
Pubmed ID
Book ISBNs
978-1-4939-6971-5, 978-1-4939-6972-2
Authors

Yashas Rajendra, Sowmya Balasubramanian, David L. Hacker

Editors

Paula Meleady

Abstract

We describe a one-liter transfection of suspension-adapted Chinese hamster ovary (CHO-DG44) cells using polyethyleneimine (PEI) for DNA delivery. The method involves transfection at a high cell density (5 × 10(6) cells/mL) by direct addition of plasmid DNA (pDNA) and PEI to the culture and subsequent 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).

Mendeley readers

Mendeley readers

The data shown below were compiled from readership statistics for 15 Mendeley readers of this research output. Click here to see the associated Mendeley record.

Geographical breakdown

Country Count As %
Unknown 15 100%

Demographic breakdown

Readers by professional status Count As %
Student > Ph. D. Student 3 20%
Student > Bachelor 3 20%
Researcher 3 20%
Student > Master 2 13%
Student > Doctoral Student 1 7%
Other 0 0%
Unknown 3 20%
Readers by discipline Count As %
Biochemistry, Genetics and Molecular Biology 8 53%
Agricultural and Biological Sciences 2 13%
Materials Science 1 7%
Chemistry 1 7%
Unknown 3 20%