Heterologous Protein Production in CHO Cells Methods and Protocols

This detailed new edition explores the use of Chinese hamster ovary (CHO) cells in the production of therapeutic protein products. Beyond updates on earlier methodologies, the book also delves into the genetic manipulation of CHO cells for recombinant protein production, analysis of CHO cells using proteomic and metabolomic approaches, as well as methods for the characterization of recombinant protein products, such as glycosylation and host cell protein analysis. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. 

Authoritative and up-to-date, Heterologous Protein Production in CHO Cells: Methods and Protocols, Second Edition is an ideal guide for researchers working to enhance and accelerate CHO productive capabilities in the coming decades.

Serum-Free and Protein-Free Media for the Cultivation of Recombinant Chinese Hamster Ovary Cells (CHO) Cell Lines.- Large-Scale Transient Transfection of Suspension-Adapted Chinese Hamster Ovary Cells for the Production of the Trimeric SARS-CoV-2 Spike Protein.- Splicing by Overlap Extension PCR for the Production of Fusion Proteins.- Codon and Signal Peptide Optimization to Enhance Therapeutic Antibody Production from CHO Cells.- Application of CRISPR/Cas9 Genome Editing to Improve Recombinant Protein Production in CHO Cells.- Conditional Knockdown of Endogenous MicroRNAs in CHO Cells Using TET-ON-SanDI Sponge Vectors.- MiRNA Chaining for Efficient Stable Overexpression to Improve Protein Quantity and Quality in CHO Cells.- Anti-Apoptosis Engineering for Improved Protein Production from CHO Cells.- The ‘Omics Revolution in CHO Biology: Roadmap to Improved CHO Productivity.- Recent Advancements in Proteomic Sample Preparation from Recombinant Chinese Hamster Ovary Cells.- Separation and Purification of CHO Secretome and Extracellular Vesicles for Proteome Analysis.- Phosphopeptide Enrichment and LC-MS/MS Analysis to Study the Phosphoproteome of Recombinant Chinese Hamster Ovary Cells.- LC-MS/MS Analysis to Study the Ubiquitin-Modified Proteome of Recombinant Chinese Hamster Ovary Cells.- Metabolomics of Chinese Hamster Ovary Cells.- Analytical Tools for HCP Detection, Identification, and Quantitation.- Quantitative High-Throughput and High-Sensitivity Glycoprofile Analysis of Therapeutic Glycoproteins Using HILIC-FLD and ESI-MS.