Hacker Recombinant Protein Expression in Mammalian Cells

Methods and Protocols
Erscheinungsjahr 2018
ISBN: 978-1-4939-8730-6
Verlag: Springer US
Format: PDF
Kopierschutz: 1 - PDF Watermark

E-Book, Englisch, Band 1850, 311 Seiten, eBook

Reihe: Methods in Molecular Biology

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.

Zielgruppe

Professional/practitioner

Autoren/Hrsg.

Hacker, David L.

Inhaltsverzeichnis

Transient Gene Expression in Suspension HEK293-EBNA1 Cells.- Transient Expression of Recombinant Membrane-eGFP Fusion Proteins in HEK293 Cells.- PEI-Mediated Transient Gene Expression in CHO Cells.- Stable Expression by Lentiviral Transduction of Cells.- Inducible Protein Production in 293 Cells using the piggyBac Transposon System.- Recombinant CHO Cell Pool Generation using piggyBac Transposon System.- Genome Engineering of Hybridomas to Generate Stable Cell Lines for Antibody Expression.- Protein Expression via Transient Transfection of Mammalian Cells in a WAVE Bioreactor.- CHO and HEK293 Cell Cultivation and Transfection in Single-Use Orbitally Shaken Bioreactors.- Bench-Scale Stirred-Tank Bioreactor for Recombinant Protein Production in Chinese Hamster Ovary Cells in Suspension.- Continuous and Integrated Expression and Purification of Recombinant Antibodies.- High Throughput Transfection of HEK293 Cells for Transient ProteinProduction.- Microfluidic Transfection for High-Throughput Mammalian Protein Expression.- Genome Wide High Throughput RNAi Screening for Identification of Genes Involved in Protein Production.- Targeting miRNAs with CRISPR/Cas9 to Improve Recombinant Protein Production of CHO Cells.- Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells.- Scalable Production and Purification of Adeno-Associated Viral Vectors (AAV) .- Considerations in the Use of Codon Optimization for Recombinant Protein Expression.- Versatile Cell-free Protein Synthesis Systems based on Chinese Hamster Ovary Cells.