Heterologous Protein Production in CHO Cells

Heterologous Protein Production in CHO Cells : Methods and Protocols

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Description

This volume discusses protocols that cover genetic manipulation of Chinese hamster ovary (CHO) cells for recombinant protein production, and protocols for the characterization of CHO cells using 'omic approaches. This book also explores methods that discuss the genome editing tool, CRISPR/Cas9, and the characterization of recombinant protein products, such as glycosylation and host cell protein analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Comprehensive and cutting-edge, Heterologous Protein Production in CHO Cells: Methods and Protocols is a valuable resource for scientists and researchers who are interested in further studying cell production in CHO cells.
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Product details

  • Hardback | 252 pages
  • 178 x 254 x 16mm | 917g
  • Totowa, NJ, United States
  • English
  • 1st ed. 2017
  • 30 Tables, color; 28 Illustrations, color; 7 Illustrations, black and white; X, 252 p. 35 illus., 28 illus. in color.
  • 1493969714
  • 9781493969715
  • 1,252,700

Back cover copy

This volume discusses protocols that cover genetic manipulation of Chinese hamster ovary (CHO) cells for recombinant protein production, and protocols for the characterization of CHO cells using 'omic approaches. This book also explores methods that discuss the genome editing tool, CRISPR/Cas9, and the characterization of recombinant protein products, such as glycosylation and host cell protein analysis. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Comprehensive and cutting-edge, Heterologous Protein Production in CHO Cells: Methods and Protocols is a valuable resource for scientists and researchers who are interested in further studying cell production in CHO cells.
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Table of contents

Preface...
Table of Contents...
Contributing Authors...



1. Strategies and Considerations for Improving Expression of 'Difficult to Express' Proteins in CHO Cells


Christina S. Alves and Terrence M. Dobrowsky



2. Glycoengineering of CHO Cells to Improve Product Quality


Qiong Wang, Bojiao Yin, Cheng-Yu Chung, and Michael J. Betenbaugh



3. Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension


Yashas Rajendra, Sowmya Balasubramanian, and David L. Hacker



4. Cloning of Single-Chain Antibody Variants by Overlap-Extension PCR for Evaluation of Antibody Expression in Transient Gene Expression


Patrick Mayrhofer and Renate Kunert



5. Anti-Apoptosis Engineering for Improved Protein Production from CHO Cells


Eric Baek, Soo Min Noh, and Gyun Min Lee



6. Conditional Knockdown of Endogenous MicroRNAs in CHO Cells using TET-ON-SanDI Sponge Vectors


Alan Costello, Nga Lao, Martin Clynes, and Niall Barron



7. Application of CRISPR/Cas9 Genome Editing to Improve Recombinant Protein Production in CHO Cells
Lise Marie Grav, Karen Julie la Cour Karottki, Jae Seong Lee, and Helene Faustrup Kildegaard



8. Improved CHO Cell Line Stability and Recombinant Protein Expression During Long-Term Culture


Zeynep Betts and Alan J. Dickson



9. Selection of High-Producing Clones using FACS for CHO Cell Line Development


Clair Gallagher and Paul S. Kelly



10. The 'Omics Revolution in CHO Biology: Roadmap to Improved CHO Productivity


Hussain Dahodwala and Susan T. Sharfstein



11. A Bioinformatics Pipeline for the Identification of CHO Cell Differential Gene Expression from RNA-Seq Data


Craig Monger, Krishna Motheramgari, John McSharry, Niall Barron, and Colin Clarke



12. Filter-Aided Sample Preparation (FASP) for Improved Proteome Analysis of Recombinant Chinese Hamster Ovary Cells
Orla Coleman, Michael Henry, Martin Clynes, and Paula Meleady



13. Phosphopeptide Enrichment and LC-MS/MS Analysis to Study the Phosphoproteome of Recombinant Chinese Hamster Ovary Cells
Michael Henry, Orla Coleman, Prashant, Martin Clynes, and Paula Meleady



14. Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture


Yuzhou Fan, Helene Faustrup Kildegaard, and Mikael Rordam Andersen



15. Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells


Sara Carillo, Stefan Mittermayr, Amy Farrell, Simone Albrecht, and Jonathan Bones



16. Characterization of Host Cell Proteins (HCPs) in CHO Cell Bioprocesses


Catherine E.M. Hogwood, Lesley M. Chiverton, and C. Mark Smales
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