Recombinant Protein Expression in Mammalian Cells - Methods and Protocols

This fully updated volume explores notable developments in the field of mammalian cell-based recombinant protein production. Beginning with methods for transient recombinant protein production, the book continues with methods for stable cell pool generation, protein production using stable clonal cell lines, as well as high-throughput screening technologies for characterizing transient cell surface protein ectodomain expression and for identifying host genes involved in protein production. 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 practical, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols, Second Edition serves as an ideal guide for researchers investigatingprotein structure and function and accelerating the discovery of new therapeutic proteins.Table of ContentPEI-Mediated Transient Gene Expression in CHO Cells.- Transient Co-Expression of Membrane Protein Complexes in Mammalian Cells.- The Use of Baculovirus-Mediated Gene Expression in Mammalian Cells for Recombinant Protein Production.- Production and Purification of Adeno-Associated Viral Vectors (AAV) Using Orbitally Shaken HEK293 Cells.- Protein Expression via Transient Transfection of Mammalian Cells in a WAVE Bioreactor.- Small-Scale Cultivation and Transfection of ExpiCHO and HEK293E Cells in Single-Use Orbitally Shaken Bioreactors.- Recombinant Protein Production from Stable CHO Cell Pools.- A Versatile Method for Inducible Protein Production in 293 Cells via the PiggyBac Transposon System.- Recombinant CHO Cell Pool Generation Using PiggyBac Transposon System.- Stable Expression by Lentiviral Transduction of Cells.- Expression of Multispecific Antibodies.- Expression of Bispecific Antibodies Using Cellular Display.- Generation of Monoclonal Chinese hamster Ovary Cell Lines Using DISPENCELL-S3.- Enhancing Cell Line Stability by CRISPR/Cas9-Mediated Site-Specific Integration Based on Histone Modifications.- Bench-Scale Stirred-Tank Bioreactor for Recombinant Protein Production in Chinese Hamster Ovary (CHO) Cells in Suspension.- Application of the CRISPR/Cas9 Gene Editing Method for Modulating Antibody Fucosylation in CHO Cells.- Development of Responsive Promoters and Their Utilization for Stable CHO Sensor Cell Lines.- CRISPR-Deletion of miR-27 Impacts Recombinant Protein Production in CHO Cells.- High-Throughput Protein Expression Screening of Cell Surface Protein Ectodomains.- Genome Wide High Throughput RNAi Screening for Identification of Genes Involved in Protein Production.- Purification and Quality Control of Recombinant Proteins Expressed in Mammalian Cells: A Practical Review.This fully updated volume explores notable developments in the field of mammalian cell-based recombinant protein production. Beginning with methods for transient recombinant protein production, the book continues with methods for stable cell pool generation, protein production using stable clonal cell lines, as well as high-throughput screening technologies for characterizing transient cell surface protein ectodomain expression and for identifying host genes involved in protein production. 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 practical, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols, Second Edition serves as an ideal guide for researchers investigatingprotein structure and function and accelerating the discovery of new therapeutic proteins.Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts