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This detailed volume presents a series of protocols dealing with different aspects of inclusion body (IB) processing, from cloning procedures to purification of refolded product. Commencing with chapters on upstream processing, looking into different expression strategies for IB production, the book continues with downstream applications, highlighting early protein purification and subsequent analytics, as well as success stories of IB-based processes. 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, Inclusion Bodies: Methods and Protocols serves as an ideal resource for facilitating diverse aspects of IB processing.
List of contents
Inclusion Bodies: Status Quo and Perspectives.- Thermoinducible E. coli for Recombinant Protein Production in Inclusion Bodies.- High-Throughput Expression of Inclusion Bodies on an Automated Platform.- Design, Production, and Characterization of Catalytically-Active Inclusion Bodies.- Bidirectional Promoter Libraries Enable the Balanced Co-Expression of Two Target Genes in E. coli.- Inclusion Body Production in Fed-Batch and Continuous Cultivation.- Avoiding the All-or-None Response in Gene Expression during E. coli Continuous Cultivation Based on the On-Line Monitoring of Cell Phenotypic Switching Dynamics.- Optimization of Inclusion Body Formation and Purification in Multi-Well Plates.- High Pressure Homogenization for Inclusion Body Isolation.- Development of Solubilization and Refolding Buffers.- Unit Operation-Spanning Investigation of the Redox System.- Using High Pressure and Alkaline pH for Refolding.- Intensification of Inclusion Body Processing viaTemperature-Based Refolding.- Refolding of Proteins Expressed as Inclusion Bodies in E. coli.- Infrared Spectroscopy for Structure Analysis of Protein Inclusion Bodies.- The Purification of Heme Peroxidases from Escherichia coli Inclusion Bodies: A Success Story Shown by the Example of Horseradish Peroxidase.- Production of -Synuclein Fibrillar Specific scFv from Inclusion Bodies.- Method for Inclusion Bodies Production via E. coli Host System: rGCSF as Model Biotherapeutic Protein.- Antimicrobial Applications of Inclusion Bodies.
