Recombinant protein expression in microbial systems PDF Download
Are you looking for read ebook online? Search for your book and save it on your Kindle device, PC, phones or tablets. Download Recombinant protein expression in microbial systems PDF full book. Access full book title Recombinant protein expression in microbial systems by Eduardo A. Ceccarelli. Download full books in PDF and EPUB format.
Author: Eduardo A. Ceccarelli
Publisher: Frontiers E-books
ISBN: 2889192946
Category : Biotechnology
Languages : en
Pages : 103
Get Book
Book Description
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.
Author: Eduardo A. Ceccarelli
Publisher: Frontiers E-books
ISBN: 2889192946
Category : Biotechnology
Languages : en
Pages : 103
Get Book
Book Description
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.
Author: Gerd Gellissen
Publisher: John Wiley & Sons
ISBN: 3527604413
Category : Science
Languages : en
Pages : 429
Get Book
Book Description
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."
Author: Varsha Gupta
Publisher: Springer
ISBN: 9811008752
Category : Technology & Engineering
Languages : en
Pages : 527
Get Book
Book Description
This book explores the journey of biotechnology, searching for new avenues and noting the impressive accomplishments to date. It has harmonious blend of facts, applications and new ideas. Fast-paced biotechnologies are broadly applied and are being continuously explored in areas like the environmental, industrial, agricultural and medical sciences. The sequencing of the human genome has opened new therapeutic opportunities and enriched the field of medical biotechnology while analysis of biomolecules using proteomics and microarray technologies along with the simultaneous discovery and development of new modes of detection are paving the way for ever-faster and more reliable diagnostic methods. Life-saving bio-pharmaceuticals are being churned out at an amazing rate, and the unraveling of biological processes has facilitated drug designing and discovery processes. Advances in regenerative medical technologies (stem cell therapy, tissue engineering, and gene therapy) look extremely promising, transcending the limitations of all existing fields and opening new dimensions for characterizing and combating diseases.
Author: Brigitte Gasser
Publisher: Humana Press
ISBN: 9781493990238
Category : Science
Languages : en
Pages : 431
Get Book
Book Description
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.
Author: Otto-Wilhelm Merten
Publisher: Springer Science & Business Media
ISBN: 9780792371373
Category : Medical
Languages : en
Pages : 434
Get Book
Book Description
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.
Author: A. van Broekhoven
Publisher: Springer Science & Business Media
ISBN: 0306468859
Category : Technology & Engineering
Languages : en
Pages : 454
Get Book
Book Description
The present book entitled “Novel Frontiers in the Production of Compounds for Biomedical Uses” can perhaps be placed in its best perspective by the Shakespearean character in The Tempest who exclaimed" What’s past is prologue”. Indeed, this compilation of some of the outstanding presentations in the field of biomedicine made at th the 9 European Congress on Biotechnology (Brussels, Belgium, July 11-15, 1999) not only reflects the achievements of the recent past, but provides a privileged glimpse of the biotechnology that is emerging in the first decade of the new Millennium. It is becoming increasingly apparent that biotechnology is offering biomedicine novel approaches and solutions to develop a sorely needed new generation of biopharmaceuticals. This is all the more necessary because in recent years, new diseases have emerged with extraordinary lethality in all corners of the globe, while age-related chronic illnesses have filled the gap wherever biomedicine has made successful inroads. The rise of antibiotic resistance also poses major threats to public health. Thus, as disease patterns evolve, the rational development of new drugs is becoming urgent, not only for the clinical outcome of patients, but also in optimising the allocation of scarce health care resources through the use of cost-effective productions methods. It is in response to all these challenges that biotechnology offers new strategies that go beyond the more traditional approaches. By the mid-1990’s, the number of recombinant products approved annually for therapeutic use reached double digits. With the advent of the genomics revolution.
Author:
Publisher: Academic Press
ISBN: 0323901476
Category : Science
Languages : en
Pages : 462
Get Book
Book Description
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
Author: Elena García-Fruitós
Publisher: Humana
ISBN: 9781493946754
Category : Science
Languages : en
Pages : 0
Get Book
Book Description
With insolubility proving to be one of the most crippling bottlenecks in the protein production and purification process, this volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Insoluble Proteins: Methods and Protocols includes chapters that describe not only the recombinant protein production in different expression systems but also different purification and characterization methods to finally obtain these difficult-to-obtain proteins. Beginning with protein production methods using both prokaryotic and eukaryotic expression systems, the book continues with purification protocols using insoluble proteins, the characterization of insoluble proteins, as well as a general overview of interesting applications of insoluble 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. Comprehensive and practical, Insoluble Proteins: Methods and Protocols aims to provide the scientific community with detailed and reliable state-of-the-art protocols that are used in order to successfully produce and purify recombinant proteins prone to aggregate.
Author: Joseph M. Fernandez
Publisher: Elsevier
ISBN: 0080532357
Category : Science
Languages : en
Pages : 480
Get Book
Book Description
Gene Expression Systems: Using Nature for the Art of Expression offers detailed information on a wide variety of gene expression systems from an array of organisms. It describes several different types of expression systems including transient, stable, viral, and transgenic systems. Each chapter is written by a leader in the field. The book includes timelines and examples for each expression system, and provides an overview of the future of recombinant protein expression. Provides detailed information on expression systems Covers a variety of promoters and host organisms enabling researchers to tailor protocols to their specific needs Includes timelines and examples Compares pros and cons of each method
Author: Michael Dyson
Publisher: Scion Publishing Ltd
ISBN: 1907904409
Category : Science
Languages : en
Pages : 311
Get Book
Book Description
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
