Advanced Technologies for Protein Complex Production and Characterization

Advanced Technologies for Protein Complex Production and Characterization
Author: M. Cristina Vega
Publisher: Springer
Total Pages: 378
Release: 2016-05-10
Genre: Science
ISBN: 3319272160
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This book presents advanced expression technologies for the production of protein complexes. Since complexes lie at the heart of modern biology, the expression, purification, and characterization of large amounts of high-quality protein complexes is crucial for the fields of biomedicine, biotechnology, and structural biology. From co-expression in E. coli, yeast, mammalian and insect cells to complex reconstitution from individual subunits, this book offers useful insights and guidance for successful protein expressionists. Across several sections readers will discover existing opportunities for the production of protein complexes in bacterial systems (including membrane proteins and cell-free co-expression), methylotrophic and non-methylotrophic yeasts, protozoa (Leishmania terantolae and Dictyostelium discoideum), baculovirus-infected insect cells, mammalian cells, plants and algae. Complex reconstitution from individually purified subunits or subcomplexes is discussed as a complementary strategy. A last section introduces briefly some of the biophysical and structural characterization techniques for macromolecular complexes using state-of-the-art solution scattering and nuclear magnetic resonance. This work is a guided tour over some of the most powerful and successful protein expression technologies, with a focus on co-expression and high-throughput applications. It is addressed to everyone interested in the production and characterization of macromolecular complexes, from university students who want an accessible description of the major co-expression systems to researchers in biomedicine and the life sciences seeking for an up-to-date survey of available technologies.

Advanced Technologies for Protein Complex Production and Characterization

Advanced Technologies for Protein Complex Production and Characterization
Author: M. Cristina Vega
Publisher: Springer
Total Pages: 0
Release: 2024-03-22
Genre: Science
ISBN: 9783031521928
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This edited volume discusses the identification, discovery, characterization, structure determination and modeling of multicomponent macromolecular complexes, and as such, it fully complements the first volume (ISBN 978-3-319-27214-6), which targeted methods of recombinant production of protein complexes. This book is divided in 8 sections offering a selection of technologies widely used in the characterization of protein and protein-nucleic acid complexes for different purposes and at different scales. From native electrophoresis methods, that are accessible to any reasonably well-equipped laboratory, to the sophisticated setup required for structure determination by cryo-electron microscopy or X-ray crystallography, this book contains a wide variety of clearly explained analytic and preparative approaches, connected with the production techniques developed on the previous volume. The readers will find an integral connection between this book and the first volume, which ensures a comprehensive and updated discussion of the main topics of the discipline. Taken together, these volumes constitute a cohesive and authoritative source of the research on multicomponent macromolecular complexes. In here, we focus on characterization of protein complexes in the broadest sense, which is not typically covered in other sources. Moreover, all chapters are carefully written by world renowned scientists and active researchers, making this volume ideal, not only as a reference source, but also as a companion book for the daily laboratory work. This book is aimed for a wide range of scientists, from science students to experienced researchers, working on protein and protein-nucleic acid complexes, who need a thorough understanding of protein production and complex characterization.

Advanced Technologies for Protein Complex Production and Characterization

Advanced Technologies for Protein Complex Production and Characterization
Author: M. Cristina Vega
Publisher: Springer Nature
Total Pages: 217
Release: 2024
Genre: Biochemistry
ISBN: 3031521935
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Zusammenfassung: This edited volume discusses the identification, discovery, characterization, structure determination and modeling of multicomponent macromolecular complexes, and as such, it fully complements the first volume (ISBN 978-3-319-27214-6), which targeted methods of recombinant production of protein complexes. This book is divided in 8 sections offering a selection of technologies widely used in the characterization of protein and protein-nucleic acid complexes for different purposes and at different scales. From native electrophoresis methods, that are accessible to any reasonably well-equipped laboratory, to the sophisticated setup required for structure determination by cryo-electron microscopy or X-ray crystallography, this book contains a wide variety of clearly explained analytic and preparative approaches, connected with the production techniques developed on the previous volume. The readers will find an integral connection between this book and the first volume, which ensures a comprehensive and updated discussion of the main topics of the discipline. Taken together, these volumes constitute a cohesive and authoritative source of the research on multicomponent macromolecular complexes. In here, we focus on characterization of protein complexes in the broadest sense, which is not typically covered in other sources. Moreover, all chapters are carefully written by world renowned scientists and active researchers, making this volume ideal, not only as a reference source, but also as a companion book for the daily laboratory work. This book is aimed for a wide range of scientists, from science students to experienced researchers, working on protein and protein-nucleic acid complexes, who need a thorough understanding of protein production and complex characterization.

Fundamentals of Recombinant Protein Production, Purification and Characterization

Fundamentals of Recombinant Protein Production, Purification and Characterization
Author: Deepti Yadav
Publisher: Elsevier
Total Pages: 307
Release: 2024-09-27
Genre: Science
ISBN: 032398584X
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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

Approaches to the Purification, Analysis and Characterization of Antibody-Based Therapeutics

Approaches to the Purification, Analysis and Characterization of Antibody-Based Therapeutics
Author: Allan Matte
Publisher: Elsevier
Total Pages: 222
Release: 2020-09-07
Genre: Business & Economics
ISBN: 0081030193
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Approaches to the Purification, Analysis and Characterization of Antibody-Based Therapeutics provides the interested and informed reader with an overview of current approaches, strategies and considerations relating to the purification, analytics and characterization of therapeutic antibodies and related molecules. While there are obviously other books published in and around this subject area, they seem to be either older (c.a. year 2000 publication date) or are more limited in scope. The book will include an extensive bibliography of the published literature in the respective areas covered. It is not, however, intended to be a how-to methods book. Covers the vital new area of R&D on therapeutic antibodies Written by leading scientists and researchers Up-to-date coverage and includes a detailed bibliography

Insoluble Proteins

Insoluble Proteins
Author: Elena García-Fruitós
Publisher: Humana Press
Total Pages: 0
Release: 2014-12-02
Genre: Science
ISBN: 9781493922048
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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.

Recombinant Protein Expression: Prokaryotic hosts and cell-free systems

Recombinant Protein Expression: Prokaryotic hosts and cell-free systems
Author:
Publisher: Academic Press
Total Pages: 462
Release: 2021-10-29
Genre: Science
ISBN: 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

Metabolic Engineering for Bioactive Compounds

Metabolic Engineering for Bioactive Compounds
Author: Vipin Chandra Kalia
Publisher: Springer
Total Pages: 413
Release: 2017-10-07
Genre: Medical
ISBN: 9811055114
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This book comprehensively discusses the latest research in the area of metabolic engineering. Metabolic engineering solutions for bioactive compounds are now being derived by means of heterologous gene expression, in a wide range of organisms. The book provides an overview of the model systems being employed for metabolic manipulation to yield bioactive molecules, such as single-cell proteins, antibody generation, metabolites, proteases, chaperones, therapeutic proteins, nanomaterials, polymeric conjugates, dendrimers and nanoassemblies, Escherichia coli, Agrobacterium, Saccharomyces cerevisiae and cell lines, etc. In addition, it shares insights into the scope of these methods in the areas of prevention, diagnosis and treatment of diseases, e.g. immunotherapy for curing various diseases like cancer, allergies, autoimmune diseases, etc.

High-Throughput Protein Production and Purification

High-Throughput Protein Production and Purification
Author: Renaud Vincentelli
Publisher: Humana
Total Pages: 537
Release: 2019-07-05
Genre: Science
ISBN: 9781493996230
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This book compiles key protocols instrumental to the study of high-throughput protein production and purification which have been refined and simplified over the years and are now ready to be transferred to any laboratory. Beginning with a section covering general procedures for high-throughput protein production, the volume continues with high-throughput protocols adapted to the production of specific protein families, as well as an extensive section on protocols combining high-throughput protein production and their micro-characterization. Written for the highly successful Methods in Molecular Biology series, chapters in this book 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 practical, High-Throughput Protein Production and Purification: Methods and Protocols serves biochemists ranging from engineers, PhD students and post-doctoral fellows, to the heads of protein expression facilities and researchers, in pursuing this vital area of study.

Advanced Methods in Structural Biology

Advanced Methods in Structural Biology
Author: Ângela Sousa
Publisher: Springer Nature
Total Pages: 462
Release: 2023-04-24
Genre: Science
ISBN: 1071631470
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This volume explores the latest integrated bioprocesses and technologies used to study the production of the target recombinant protein of therapeutic or diagnostic interest; its isolation, purification, and stabilization; and the bio-interaction and structural analyses. The chapters in this book are organized into four parts. Part One covers production methods of soluble and membrane proteins in prokaryotic and eukaryotic expression systems, such as Lactococcus lactis and Escherichia coli. Part Two describes traditional and novel approaches for recombinant protein purification and stabilization, and buffers and additives. Part Three discusses automated methods in structural biology based on in silico approaches; and Part Four provides examples of advanced protein investigation methodologies to assess structural analysis such as high-throughput protein crystallization and time-resolved serial crystallography. 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. Cutting-edge and comprehensive, Advanced Methods in Structural Biology is a valuable resource to those in academia (i.e., graduate students and postdoctoral researchers) and researchers in the pharmaceutical industry who wish to learn more about this developing field. Chapter 5 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.