Description
Garner G. Moulton has a Masters of Science in Biology from the Program of Cell and Molecular Biology at San Diego State University. He has been working in biotech and non-profit research for over 25 years, and has focussed specifically on protein purification and fermentation process development over the last 17 years. During the past 4 years he has been employed by the Infectious Disease Research Institute (IDRI) in Seattle, WA, USA, where he is a principal research associate in charge of the Process Development Department. He has developed many robust fermentation processes that can be adapted to many different recombinant products without having to re-optimize conditions of growth and expression. Numerous projects have been successfully transferred to a contract manufacturer for scale-up production and use in clinical trials. 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
