Recombinant Protein Expression in Mammalian Cells: Methods and Protocols

ISBN-13: 9781493993772 / Angielski / Miękka / 2019 / 311 str.

David L. Hacker

Spis treści

1.         Transient Gene Expression in Suspension HEK293-EBNA1 Cells

Denis L’Abbé, Louis Bisson, Christian Gervais, Eric Grazzini and Yves Durocher

 

2.         Transient Expression of Recombinant Membrane-eGFP Fusion Proteins in HEK293 Cells

Joanna Pieprzyk, Samuel Pazicky and Christian Löw

 

3.         PEI-Mediated Transient Gene Expression in CHO Cells

Yashas Rajendra

 

4.         Stable Expression by Lentiviral Transduction of Cells

Natascha Gödecke, Hansjörg Hauser and Dagmar Wirth

 

5.         Inducible Protein Production in 293 Cells using the piggyBac Transposon System

Iacovos P. Michael and Andras Nagy

 

6.         Recombinant CHO Cell Pool Generation using piggyBac Transposon System

Sowmya Balasubramanian

 

7.         Genome Engineering of Hybridomas to Generate Stable Cell Lines for Antibody Expression

Cristina Parola, Derek Mason, Andreas Zingg, Daniel Neumeier and Sai T. Reddy

 

8.         Protein Expression via Transient Transfection of Mammalian Cells in a WAVE Bioreactor

David I. Fisher

 

9.         CHO and HEK293 Cell Cultivation and Transfection in Single-Use Orbitally Shaken Bioreactors

David L. Hacker, Laurence Durrer and Soraya Quinche

 

10.       Bench-Scale Stirred-Tank Bioreactor for Recombinant Protein Production in Chinese Hamster Ovary Cells in Suspension

Dominique Monteil and Jeffrey Kuan

 

11.       Continuous and Integrated Expression and Purification of Recombinant Antibodies

Sebastian Vogg, Moritz Wolf and Massimo Morbidelli

 

12.       High Throughput Transfection of HEK293 Cells for Transient Protein Production

Tia A. Arena, Peter D. Harms and Athena W. Wong

 

13.       Microfluidic Transfection for High-Throughput Mammalian Protein Expression

Kristina Woodruff and Sebastian J. Maerkl

 

14.       Genome Wide High Throughput RNAi Screening for Identification of Genes Involved in Protein Production

Sarah Inwood, Michael Betenbaugh, Madhu Lal and Joseph Shiloach

 

15.       Targeting miRNAs with CRISPR/Cas9 to Improve Recombinant Protein Production of CHO Cells

Kevin Kellner, Ankur Solanki, Thomas Amann, Nga T Lao and Niall Barron

 

16.       Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells

Qiong Wang, Cheng-Yu Chung, Julian N. Rosenberg, Geng Yu and Michael J. Betenbaugh

 

17.       Scalable Production and Purification of Adeno-Associated Viral Vectors (AAV)

Daniel Blessing, Nicole Déglon and Bernard L. Schneider

 

18.       Considerations in the Use of Codon Optimization for Recombinant Protein Expression 

Vincent P. Mauro and Stephen A. Chappell

 

19.       Versatile Cell-free Protein Synthesis Systems based on Chinese Hamster Ovary Cells

Lena Thoring and Stefan Kubick

Blurb

This detailed volume explores advances in vector design, DNA delivery, cell cultivation, host cell engineering, and bioprocess optimization within the study of recombinant protein expression in mammalian cells. The majority of the protocols employ either Chinese hamster ovary cells (CHO) or human embryonic kidney 293 cells (HEK293), the workhorses of the field, as the production host;  however, the methods can be adapted to other mammalian hosts under the appropriate cell-specific conditions. 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.

Authoritative and convenient, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols aims to aid researchers in building on our knowledge of protein structure and function and to speed the discovery of new therapeutic proteins.

Chapter 19 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com