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Full 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.
Contents
Strategies and Considerations for Improving Expression of 'Difficult to Express' Proteins in CHO Cells.- Glycoengineering of CHO Cells to Improve Product Quality.- Large-Scale Transient Transfection of Chinese Hamster Ovary Cells in Suspension.- Cloning of Single-Chain Antibody Variants by Overlap-Extension PCR for Evaluation of Antibody Expression in Transient Gene Expression.- Anti-Apoptosis Engineering for Improved Protein Production from CHO Cells.- Conditional Knockdown of Endogenous MicroRNAs in CHO Cells using TET-ON-SanDI Sponge Vectors.- Application of CRISPR/Cas9 Genome Editing to Improve Recombinant Protein Production in CHO Cells.- Improved CHO Cell Line Stability and Recombinant Protein Expression During Long-Term Culture.- Selection of High-Producing Clones using FACS for CHO Cell Line Development.- The 'Omics Revolution in CHO Biology: Roadmap to Improved CHO Productivity.- A Bioinformatics Pipeline for the Identification of CHO Cell Differential Gene Expression from RNA-Seq Data.- Filter-Aided Sample Preparation (FASP) for Improved Proteome Analysis of Recombinant Chinese Hamster Ovary Cells.- Phosphopeptide Enrichment and LC-MS/MS Analysis to Study the Phosphoproteome of Recombinant Chinese Hamster Ovary Cells.- Engineer Medium and Feed for Modulating N-Glycosylation of Recombinant Protein Production in CHO Cell Culture.- Glycosylation Analysis of Therapeutic Glycoproteins Produced in CHO Cells.- Characterization of Host Cell Proteins (HCPs) in CHO Cell Bioprocesses.
