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Fundamentals of Recombinant Protein Production, Purification and Characterization

Fundamentals of Recombinant Protein Production, Purification and Characterization PDF Author: Deepti Yadav
Publisher: Elsevier
ISBN: 032398584X
Category : Science
Languages : en
Pages : 307

Book Description
Fundamentals of Recombinant Protein Production, Purification and Characterization is organized into nine chapters in a logical fashion that cover an introduction to recombinant proteins and expression in different host expression systems, extraction, purification and analysis of proteins. This important reference features protocols, along with the advantages and disadvantage of each expression hosts and characterization technique (presented in tabular format) and offers detailed coverage of all aspects of protein production and processing (upstream and downstream processing) in one place. Finally, the book ends with different characterization techniques. Production of recombinant proteins for biotechnological and therapeutic applications at a large scale is an essential need of mankind. With the huge application potential of therapeutic and industrial proteins, there has been increasing demand for effective and efficient bioprocessing strategies. Recent progress around recombinant DNA technologies and bioprocessing strategies has paved the way for efficient production of recombinant proteins. Important factors such as insolubility and cost of production need to be considered for large scale production of these recombinant proteins. - Includes step-by-step reproducible protocols while also providing updated information on the rationale and latest developments in expression systems - Can also be used as a handbook for protein expression and purification as expression systems and chromatographic methods are explained in detail - Consists of notes on troubleshooting from the eminent researchers in the field - Provides comprehensive information on protein production, purification and characterization in a single volume - Describes different purification methods for comparatively difficult to obtain proteins - Brings the topics of recombinant protein expression, purification and characterization together, thereby making it the first resource on how to solve problems with respect to upstream and downstream processing of heterologous proteins

Fundamentals of Recombinant Protein Production, Purification and Characterization

Fundamentals of Recombinant Protein Production, Purification and Characterization PDF Author: Deepti Yadav
Publisher: Elsevier
ISBN: 032398584X
Category : Science
Languages : en
Pages : 307

Book Description
Fundamentals of Recombinant Protein Production, Purification and Characterization is organized into nine chapters in a logical fashion that cover an introduction to recombinant proteins and expression in different host expression systems, extraction, purification and analysis of proteins. This important reference features protocols, along with the advantages and disadvantage of each expression hosts and characterization technique (presented in tabular format) and offers detailed coverage of all aspects of protein production and processing (upstream and downstream processing) in one place. Finally, the book ends with different characterization techniques. Production of recombinant proteins for biotechnological and therapeutic applications at a large scale is an essential need of mankind. With the huge application potential of therapeutic and industrial proteins, there has been increasing demand for effective and efficient bioprocessing strategies. Recent progress around recombinant DNA technologies and bioprocessing strategies has paved the way for efficient production of recombinant proteins. Important factors such as insolubility and cost of production need to be considered for large scale production of these recombinant proteins. - Includes step-by-step reproducible protocols while also providing updated information on the rationale and latest developments in expression systems - Can also be used as a handbook for protein expression and purification as expression systems and chromatographic methods are explained in detail - Consists of notes on troubleshooting from the eminent researchers in the field - Provides comprehensive information on protein production, purification and characterization in a single volume - Describes different purification methods for comparatively difficult to obtain proteins - Brings the topics of recombinant protein expression, purification and characterization together, thereby making it the first resource on how to solve problems with respect to upstream and downstream processing of heterologous proteins

Recombinant protein expression in microbial systems

Recombinant protein expression in microbial systems PDF Author: Eduardo A. Ceccarelli
Publisher: Frontiers E-books
ISBN: 2889192946
Category : Biotechnology
Languages : en
Pages : 103

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.

Current Protocols in Protein Science

Current Protocols in Protein Science PDF Author:
Publisher:
ISBN:
Category : Proteins
Languages : en
Pages :

Book Description


Protein Refolding

Protein Refolding PDF Author: George Georgiou
Publisher:
ISBN:
Category : SCIENCE
Languages : en
Pages : 248

Book Description
the refolding process is often the critical bottleneck in the production of high-value proteins, and recently acquired insights have yet to be translated into technological advantages. These proceedings bridge the gap between fundamental and applied studies, addressing such issues as in vivo protein folding, protein aggregation and inclusion body formation, elucidation of the folding pathway, characterization of folding intermediates, and practical considerations in protein renaturation. The symposium was part of the 199th ACS National Meeting, Boston, April 1990. Annotation copyrighted by Book News, Inc., Portland, OR

Pharmaceutical Biotechnology

Pharmaceutical Biotechnology PDF Author: Daan J. A. Crommelin
Publisher: CRC Press
ISBN: 9780415285018
Category : Medical
Languages : en
Pages : 456

Book Description
The field of pharmaceutical biotechnology is evolving rapidly. A whole new arsenal of protein pharmaceuticals is being produced by recombinant techniques for cancer, viral infections, cardiovascular and hereditary disorders, and other diseases. In addition, scientists are confronted with new technologies such as polymerase chain reactions, combinatorial chemistry and gene therapy. This introductory textbook provides extensive coverage of both the basic science and the applications of biotechnology-produced pharmaceuticals, with special emphasis on their clinical use. Pharmaceutical Biotechnology serves as a complete one-stop source for undergraduate pharmacists, and it is valuable for researchers and professionals in the pharmaceutical industry as well.

Fundamentals of Protein Biotechnology

Fundamentals of Protein Biotechnology PDF Author: Stein
Publisher: Routledge
ISBN: 1351446355
Category : Medical
Languages : en
Pages : 336

Book Description
Fills a gap between the existing studies of proteins, which tend to be highly technical and geared toward the practicing protein chemist, and biochemistry textbooks, which focus on general principles. Scientists cover a dozen topics by presenting fundamental principles, an overview, and the practica

Protein Analysis and Purification

Protein Analysis and Purification PDF Author: I.M. Rosenberg
Publisher: Springer Science & Business Media
ISBN: 1475711085
Category : Mathematics
Languages : en
Pages : 453

Book Description
This book is designed to be a practical progression of experimental techniques an investigator may follow when embarking on a biochemical project. The protocols may be performed in the order laid out or may be used inde pendently. The aim of the book is to assist a wide range of researchers. from the novice to the frustrated veteran, in the choice and design of experiments that are to be performed to provide answers to specific questions. The manual describes standard techniques that have been shown to work, as well as some newer ones that are beginning to prove important. By following the promi nently numbered steps. you can work your way through any protocol. whether it's a new technique or a task you've done before for which you need a quick review or updated methodology. This manual will assist the experimentalist in designing properly controlled experiments. There will be no advice for dealing with specific pieces of equip ment other than encouragement to read the manual, if you can find it. Through out all manipulations try to be objective. Be on the lookout for unexpected findings. You will learn the most from unexpected results. and they are often the beginning of the next project. It is never possible to record too much in your lab notebook. Do not get discouraged. Remember, things will not always run smoothly.

Guide to Protein Purification

Guide to Protein Purification PDF Author: Richard R Burgess
Publisher: Academic Press
ISBN: 0080923178
Category : Science
Languages : en
Pages : 915

Book Description
Guide to Protein Purification, Second Edition provides a complete update to existing methods in the field, reflecting the enormous advances made in the last two decades. In particular, proteomics, mass spectrometry, and DNA technology have revolutionized the field since the first edition's publication but through all of the advancements, the purification of proteins is still an indispensable first step in understanding their function. This volume examines the most reliable, robust methods for researchers in biochemistry, molecular and cell biology, genetics, pharmacology and biotechnology and sets a standard for best practices in the field. It relates how these traditional and new cutting-edge methods connect to the explosive advancements in the field. This "Guide to" gives imminently practical advice to avoid costly mistakes in choosing a method and brings in perspective from the premier researchers while presents a comprehensive overview of the field today. - Gathers top global authors from industry, medicine, and research fields across a wide variety of disciplines, including biochemistry, genetics, oncology, pharmacology, dermatology and immunology - Assembles chapters on both common and less common relevant techniques - Provides robust methods as well as an analysis of the advancements in the field that, for an individual investigator, can be a demanding and time-consuming process

Flow Cytometry

Flow Cytometry PDF Author: Alice Longobardi Givan
Publisher: John Wiley & Sons
ISBN: 1118688392
Category : Science
Languages : en
Pages : 309

Book Description
Flow cytometry continually amazes scientists with its ever-expanding utility. Advances in flow cytometry have opened new directions in theoretical science, clinical diagnosis, and medical practice. The new edition of Flow Cytometry: First Principles provides a thorough update of this now classic text, reflecting innovations in the field while outlining the fundamental elements of instrumentation, sample preparation, and data analysis. Flow Cytometry: First Principles, Second Edition explains the basic principles of flow cytometry, surveying its primary scientific and clinical applications and highlighting state-of-the-art techniques at the frontiers of research. This edition contains extensive revisions of all chapters, including new discussions on fluorochrome and laser options for multicolor analysis, an additionalsection on apoptosis in the chapter on DNA, and new chapters onintracellular protein staining and cell sorting, including high-speed sorting and alternative sorting methods, as well as traditional technology. This essential resource: Assumes no prior knowledge of flow cytometry Progresses with an informal, engaging lecture style from simpleto more complex concepts Offers a clear introduction to new vocabulary, principles of instrumentation, and strategies for data analysis Emphasizes the theory relevant to all flow cytometry, with examples from a variety of clinical and scientific fields Flow Cytometry: First Principles, Second Edition provides scientists, clinicians, technologists, and students with the knowledge necessary for beginning the practice of flow cytometry and for understanding related literature.

Insoluble Proteins

Insoluble Proteins PDF Author: Elena García-Fruitós
Publisher: Humana Press
ISBN: 9781493922048
Category : Science
Languages : en
Pages : 0

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.