Heterologous Protein Production in Cho Cells: Methods and Protocols

ISBN-13: 9781493969715 / Angielski / Twarda / 2017 / 252 str.

Paula Meleady

Opis

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.

Spis treści

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 Rørdam 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