Recombinant protein expression in microbial systems
Recombinant protein expression in microbial systems

Author: Eduardo A. Ceccarelli

Publisher: Frontiers E-books

Published: 2014-10-02

Total Pages: 103

ISBN-13: 2889192946

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With the advent of recombinant DNA technology, expressing heterologous proteins in microorganisms rapidly became the method of choice for their production at laboratory and industrial scale. Bacteria, yeasts and other hosts can be grown to high biomass levels efficiently and inexpensively. Obtaining high yields of recombinant proteins from this material was only feasible thanks to constant research on microbial genetics and physiology that led to novel strains, plasmids and cultivation strategies. Despite the spectacular expansion of the field, there is still much room for progress. Improving the levels of expression and the solubility of a recombinant protein can be quite challenging. Accumulation of the product in the cell can lead to stress responses which affect cell growth. Buildup of insoluble and biologically inactive aggregates (inclusion bodies) lowers the yield of production. This is particularly true for obtaining membrane proteins or high-molecular weight and multi-domain proteins. Also, obtaining eukaryotic proteins in a prokaryotic background (for example, plant or animal proteins in bacteria) results in a product that lack post-translational modifications, often required for functionality. Changing to a eukaryotic host (yeasts or filamentous fungi) may not be a proper solution since the pattern of sugar modifications is different than in higher eukaryotes. Still, many advances in the last couple of decades have provided to researchers a wide variety of strategies to maximize the production of their recombinant protein of choice. Everything starts with the careful selection of the host. Be it bacteria or yeast, a broad list of strains is available for overcoming codon use bias, incorrect disulfide bond formation, protein toxicity and lack of post-translational modifications. Also, a huge catalog of plasmids allows choosing for different fusion partners for improving solubility, protein secretion, chaperone co-expression, antibiotic resistance and promoter strength. Next, controlling culture conditions like temperature, inducer and media composition can bolster recombinant protein production. With this Research Topic, we aim to provide an encyclopedic account of the existing approaches to the expression of recombinant proteins in microorganisms, highlight recent discoveries and analyze the future prospects of this exciting and ever-growing field.

Production of Recombinant Proteins
Production of Recombinant Proteins

Author: Gerd Gellissen

Publisher: John Wiley & Sons

Published: 2006-03-06

Total Pages: 429

ISBN-13: 3527604413

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While the choices of microbial and eukaryotic expression systems for production of recombinant proteins are many, most researchers in academic and industrial settings do not have ready access to pertinent biological and technical information since it is normally scattered throughout the scientific literature. This book closes the gap by providing information on the general biology of the host organism, a description of the expression platform, a methodological section -- with strains, genetic elements, vectors and special methods, where applicable -- as well as examples of proteins produced with the respective platform. The systems thus described are well balanced by the inclusion of three prokaryotes (two Gram-negatives and one Gram-positive), four yeasts, two filamentous fungi and two higher eukaryotic cell systems -- mammalian and plant cells. Throughout, the book provides valuable practical and theoretical information on the criteria and schemes for selecting the appropriate expression platform, the possibility and practicality of a universal expression vector, and on comparative industrial-scale fermentation, with the production of a recombinant Hepatitis B vaccine chosen as an industrial example. With a foreword by Herbert P. Schweizer, Colorado State University, USA: "As a whole, this book is a valuable and overdue resource for a varied audience. It is a practical guide for academic and industrial researchers who are confronted with the design of the most suitable expression platform for their favorite protein for technical or pharmaceutical purposes. In addition, the book is also a valuable study resource for professors and students in the fields of applied biology and biotechnology."

Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology
Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology

Author: Otto-Wilhelm Merten

Publisher: Springer Science & Business Media

Published: 2001-11-30

Total Pages: 434

ISBN-13: 9780792371373

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The general field of fundamental and applied biotechnology becomes increasingly important for the production of biologicals for human and veterinary use, by using prokaryotic and eukaryotic microorganisms. The papers in the present book are refereed articles compiled from oral and poster presentations from the EFB Meeting on Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology, which was organized in Semmering/A from 5th to 8th October 2000. A special feature of this meeting was the comparison of different classes of host cells, mainly bacteria, yeasts, filamentous fungi, and animal cells, which made obvious that many physiological features of recombinant protein formation, like cell nutrition, stress responses, protein folding and secretion, or genetic stability, follow similar patterns in different expression systems. This comparative aspect is by far the point of most interest because such comparisons are rarely done, and if they are done, their results are most often kept secret by the companies who generated them. Audience: Presently, a comparable book does not exist because the compiling of manuscripts from all fields of biotechnology (prokaryotic as well as eukaryotic, up to animal cell biotechnology) is not done in general. This particularity makes this book very interesting for postgraduate students and professionals in the large field of biotechnology who want to get a more global view on the current state of the expression of recombinant biologicals in different host cell systems, the physiological problems associated with the use of different expression systems, potential approaches to solve such difficulties by metabolic engineering or the use of other host cells, and the cooperation between process development and strain improvement, which is crucial for the optimisation of both the production strain and the process. This book should be in every library of an institution/organization involved in biotechnology.

Expression Systems
Expression Systems

Author: Michael Dyson

Publisher: Scion Publishing Ltd

Published: 2007-09-15

Total Pages: 311

ISBN-13: 1907904409

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1. Expression strategy (Michael Dyson) 2. Protein expression in Escherichia coli (Rosalind Kim) 3. Expression engineering of synthetic antibodies using ribosome display (Matthew DeLisa and Lydia M. Contreras Martinez) 4. Refolding proteins from inclusion bodies (Renaud Vincentelli) 5. Selection of protein variants with improved expression using GFP-derived folding and solubility reporters (Geoffrey Waldo and Stéphanie Cabantous) 6. Protein expression in the wheat germ cell-free system (Yaeta Endo and Tatsuya Sawasaki) 7. Saccharomyces cerevisiae ; A microbial eukaryotic expression system (Christine Lang) 8. Expression of proteins in Pichia pastoris (Geoff and Joan Lin-Cereghino and Wilson Leung) 9. Improved baculovirus expression vectors (Linda King, Richard Hitchman and Robert Possee) 10. Transient transfection of insect cells for rapid expression screening and protein production (Robert Novy et al.) 11. Generation of stable CHO cell lines for protein expression (Zhijian Lu et al.) 12. Transient expression in HEK293-EBNA1 cells (Yves Durocher, Roseanne Tom and Louis Bisson) 13. Nisin- and subtilin-controlled gene expression systems for Gram-positive bacteria (Oscar Kuipers and Jan Kok) 14. Protein expression using lentiviral vectors (Bernard Massie, Renald Gilbert and Sophie Broussau) 15. Expression in mammalian cells using BacMam viruses (Yu-Chen Hu and Hsiao-Ping Lee) List of suppliers;Index

Horizons in Bioprocess Engineering
Horizons in Bioprocess Engineering

Author: Ravindra Pogaku

Publisher: Springer Nature

Published: 2019-10-12

Total Pages: 354

ISBN-13: 3030290697

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This book is divided into four parts that outline the use of science and technology for applications pertaining to chemical and bioprocess engineering. The book endeavors to help academia, researchers, and practitioners to use the principles and tools of Chemical and Bioprocess Engineering in a pertinent way, while attempting to point out the novel thoughts associated with the brain storming concepts encountered. As an example, the ability to use case studies appropriately is more important, to most practitioners.

Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology
Recombinant Protein Production with Prokaryotic and Eukaryotic Cells. A Comparative View on Host Physiology

Author: Otto-Wilhelm Merten

Publisher: Springer Science & Business Media

Published: 2013-04-17

Total Pages: 396

ISBN-13: 9401597499

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More then 20 years have passed now since the first recombinant protein producing microorganisms have been developed. In the meanwhile, numerous proteins have been produced in bacteria, yeasts and filamentous fungi, as weIl as higher eukaryotic cells, and even entire plants and animals. Many recombinant proteins are on the market today, and some of them reached substantial market volumes. On the first sight one would expect the technology - including the physiology of the host strains - to be optimised in detail after a 20 year's period of development. However, several constraints have limited the incentive for optimisation, especially in the pharmaceutical industry like the urge to proceed quickly or the requirement to define the production parameters for registration early in the development phase. The additional expenses for registration of a new production strain often prohibits a change to an optimised strain. A continuous optimisation of the entire production process is not feasible for the same reasons.

Recombinant Protein Expression: Prokaryotic hosts and cell-free systems
Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

Author:

Publisher: Academic Press

Published: 2021-10-29

Total Pages: 462

ISBN-13: 0323901476

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Recombinant Protein Expression, Part A, Volume 659 in the Methods in Enzymology series, highlights new advances in the field with this new volume presenting interesting chapters on Multiplexed analysis protein: Protein interactions of polypeptides translated in Leishmania cell-free system, MultiBac system and its applications, performance and recent, Production of antibodies in Shuffle, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to enhance transcription in yeast, Designing hybrid-promoter architectures by engineering cis-acting DNA sites to deregulate transcription in yeast, Antibody or protein-based vaccine production in plants, Cell-free protein synthesis, Plant-based expression of biologic drugs, and much more. Additional sections cover the Use of native mass spectrometry to guide detergent-based rescue of non-native oligomerization by recombinant proteins, Advancing overexpression and purification of recombinant proteins by pilot optimization through tandem affinity-buffer exchange chromatography online with native mass spectrometry, Method for High-Efficiency Fed-batch cultures of recombinant Escherichia coli, Method to transfer Chinese hamster ovary (CHO) shake flask experiments to the ambr® 250, and Expression of recombinant antibodies in Leishmania tarentolae. Provides the authority and expertise of leading contributors from an international board of authors Presents the latest release in the Methods in Enzymology serial Updated release includes the latest information on Recombinant Protein Expression

Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing
Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing

Author: Ganapathy Subramanian

Publisher: John Wiley & Sons

Published: 2021-12-20

Total Pages: 404

ISBN-13: 3527347690

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Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing Explore new trends in continuous biomanufacturing with contributions from leading practitioners in the field With the increasingly widespread acceptance and investment in the ??technology, the last decade has demonstrated the utility of continuous ??processing in the pharmaceutical industry. In Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing, distinguished biotechnologist Dr. Ganapathy Subramanian delivers a comprehensive exploration of the potential of the continuous processing of biological products and discussions of future directions in advancing continuous processing to meet new challenges and demands in the manufacture of therapeutic products. A stand-alone follow-up to the editor’s Continuous Biomanufacturing: Innovative Technologies and Methods published in 2017, this new edited volume focuses on critical aspects of process intensification, process control, and the digital transformation of biopharmaceutical processes. In addition to topics like the use of multivariant data analysis, regulatory concerns, and automation processes, the book also includes: Thorough introductions to capacitance sensors to control feeding strategies and the continuous production of viral vaccines Comprehensive explorations of strategies for the continuous upstream processing of induced microbial systems Practical discussions of preparative hydrophobic interaction chromatography and the design of modern protein-A-resins for continuous biomanufacturing In-depth examinations of bioprocess intensification approaches and the benefits of single use for process intensification Perfect for biotechnologists, bioengineers, pharmaceutical engineers, and process engineers, Process Control, Intensification, and Digitalisation in Continuous Biomanufacturing is also an indispensable resource for chemical engineers seeking a one-stop reference on continuous biomanufacturing.

Recombinant Protein Production in Yeast
Recombinant Protein Production in Yeast

Author: Brigitte Gasser

Publisher: Humana Press

Published: 2019-02-09

Total Pages: 431

ISBN-13: 9781493990238

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This volume provides an overview of the main yeast production platforms currently used and future yeast cell factories for recombinant protein production. Chapters detail approaches of genetic and metabolic engineering, co-factor containing proteins and virus-like particles, glycoproteins, and post-translational modifications of proteins. 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. Authoritative and cutting-edge, Recombinant Protein Production in Yeast: Methods and Protocols aims to provide state of the art background and methods for protein producing yeast platforms, as well as case studies for special applications.

Protein Expression Technologies
Protein Expression Technologies

Author: François Baneyx

Publisher: Garland Science

Published: 2004

Total Pages: 546

ISBN-13: 9780954523251

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Advances in protein expression technologies have mushroomed in recent years. In this book current and emerging expression technologies are reviewed. Reviews of the molecular genetics of expression systems in various organisms are presented. Topics covered include: Expression of extremophilic proteins; expression in E. coli, Bacillus spp., Saccharomyces cerevisiae, and methylotrophic yeasts; insect cell expression and the baculovirus system; and Chinese Hamster Ovary (CHO) cell lines for large-scale protein production. Also covered are two emerging expression systems, Methylobacterium extorquens AM1 and Caulobacter crescentus.