Fed-batch fermentation : a practical guide to scalable recombinant protein production in escherichia coli
著者
書誌事項
Fed-batch fermentation : a practical guide to scalable recombinant protein production in escherichia coli
（Woodhead Publishing series in biomedicine, 42）
Woodhead Publishing, c2014
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注記
Includes bibliographical references and index
Also issued online
内容説明・目次
内容説明
Fed-batch Fermentation is primarily a practical guide for recombinant protein production in E. coli using a Fed-batch Fermentation process. Ideal users of this guide are teaching labs and R&D labs that need a quick and reproducible process for recombinant protein production. It may also be used as a template for the production of recombinant protein product for use in clinical trials. The guide highlights a method whereby a medium cell density - final Ods = 30-40 (A600) - Fed-batch Fermentation process can be accomplished within a single day with minimal supervision. This process can also be done on a small (2L) scale that is scalable to 30L or more. All reagents (media, carbon source, plasmid vector and host cell) used are widely available and are relatively inexpensive. This method has been used to produce three different protein products following cGMP guidelines for Phase I clinical studies.
目次
List of figures and tables
Figures
Tables
About the author
1: Introduction to fermentation
Abstract
1.1 A brief history of early fermentation and the discovery of DNA
1.2 The rise of biotechnology I
1.3 The rise of biotechnology II
2: Generation of a recombinant Escherichia coli expression system
Abstract
2.1 Plasmids
2.2 Cloning of foreign gene into plasmid
2.3 Transcription of gene into messenger RNA (mRNA)
2.4 Host cell
2.5 Transformation of E. coli
2.6 Making competent cells
2.7 Expression screening of transformed host cells
2.8 Bacterial growth preparation
2.9 Streaking and inoculating plates
2.10 Generation of a working cell bank (glycerol stock)
3: Recombinant fed-batch fermentation using Escherichia coli
Abstract
3.1 Growth kinetics of E. coli
3.2 Reactor kinetics
3.3 The bioreactor system
3.4 Set-up and performance of a 2 liter fed-batch fermentation
3.5 Analysis of fed-batch fermentation
3.6 Sample preparation for SDS PAGE
4: Escherichia coli produced recombinant protein: Soluble versus insoluble production
Abstract
4.1 Introduction
4.2 Translation from RNA into protein
4.3 The protein
4.4 Soluble protein expression in E. coli
4.5 The inclusion body
4.6 Isolation and solubilization of inclusion bodies
4.7 Ni purification of recombinant protein product
4.8 Ni purified protein
5: The future of Escherichia coli recombinant fermentation
Abstract
References
Index
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