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Protein Expression in Mammalian Cells

Protein Expression in Mammalian Cells
Author: James L Hartley
Publisher: Humana Press
Total Pages: 276
Release: 2011-10-12
Genre: Science
ISBN: 9781617793530

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Through all of the recent progress provided by high throughput DNA sequencing technologies, it has become clearer and clearer that the study of proteins and protein organelles will be the key to unlocking our ability to manipulate cells and intervene in human disease. In Protein Expression in Mammalian Cells: Methods and Protocols, expert researchers in the field present a compendium of vital techniques to further our knowledge of mammalian protein expression. Written in the highly successful Methods in Molecular BiologyTM 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 for troubleshooting and avoiding known pitfalls. Authoritative and concise, Protein Expression in Mammalian Cells: Methods and Protocols will aid scientists seeking to delve deeper into our own biology through the medium of other mammalian cells and proteins.


Recombinant Protein Expression in Mammalian Cells

Recombinant Protein Expression in Mammalian Cells
Author: David L. Hacker
Publisher: Humana Press
Total Pages: 311
Release: 2018-09-03
Genre: Science
ISBN: 9781493987290

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This detailed volume explores advances in vector design, DNA delivery, cell cultivation, host cell engineering, and bioprocess optimization within the study of recombinant protein expression in mammalian cells. The majority of the protocols employ either Chinese hamster ovary cells (CHO) or human embryonic kidney 293 cells (HEK293), the workhorses of the field, as the production host; however, the methods can be adapted to other mammalian hosts under the appropriate cell-specific conditions. 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 convenient, Recombinant Protein Expression in Mammalian Cells: Methods and Protocols aims to aid researchers in building on our knowledge of protein structure and function and to speed the discovery of new therapeutic proteins.


Cell Culture Engineering

Cell Culture Engineering
Author: Gyun Min Lee
Publisher: John Wiley & Sons
Total Pages: 436
Release: 2020-01-13
Genre: Science
ISBN: 3527343342

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Offers a comprehensive overview of cell culture engineering, providing insight into cell engineering, systems biology approaches and processing technology In Cell Culture Engineering: Recombinant Protein Production, editors Gyun Min Lee and Helene Faustrup Kildegaard assemble top class authors to present expert coverage of topics such as: cell line development for therapeutic protein production; development of a transient gene expression upstream platform; and CHO synthetic biology. They provide readers with everything they need to know about enhancing product and bioprocess attributes using genome-scale models of CHO metabolism; omics data and mammalian systems biotechnology; perfusion culture; and much more. This all-new, up-to-date reference covers all of the important aspects of cell culture engineering, including cell engineering, system biology approaches, and processing technology. It describes the challenges in cell line development and cell engineering, e.g. via gene editing tools like CRISPR/Cas9 and with the aim to engineer glycosylation patterns. Furthermore, it gives an overview about synthetic biology approaches applied to cell culture engineering and elaborates the use of CHO cells as common cell line for protein production. In addition, the book discusses the most important aspects of production processes, including cell culture media, batch, fed-batch, and perfusion processes as well as process analytical technology, quality by design, and scale down models. -Covers key elements of cell culture engineering applied to the production of recombinant proteins for therapeutic use -Focuses on mammalian and animal cells to help highlight synthetic and systems biology approaches to cell culture engineering, exemplified by the widely used CHO cell line -Part of the renowned "Advanced Biotechnology" book series Cell Culture Engineering: Recombinant Protein Production will appeal to biotechnologists, bioengineers, life scientists, chemical engineers, and PhD students in the life sciences.


Expression Systems

Expression Systems
Author: Michael Dyson
Publisher: Scion Publishing Ltd
Total Pages: 311
Release: 2007-09-15
Genre: Science
ISBN: 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


Animal Cell Culture

Animal Cell Culture
Author: Mohamed Al-Rubeai
Publisher: Springer
Total Pages: 766
Release: 2014-11-28
Genre: Medical
ISBN: 3319103202

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​Animal cells are the preferred “cell factories” for the production of complex molecules and antibodies for use as prophylactics, therapeutics or diagnostics. Animal cells are required for the correct post-translational processing (including glycosylation) of biopharmaceutical protein products. They are used for the production of viral vectors for gene therapy. Major targets for this therapy include cancer, HIV, arthritis, cardiovascular and CNS diseases and cystic fibrosis. Animal cells are used as in vitro substrates in pharmacological and toxicological studies. This book is designed to serve as a comprehensive review of animal cell culture, covering the current status of both research and applications. For the student or R&D scientist or new researcher the protocols are central to the performance of cell culture work, yet a broad understanding is essential for translation of laboratory findings into the industrial production. Within the broad scope of the book, each topic is reviewed authoritatively by experts in the field to produce state-of-the-art collection of current research. A major reference volume on cell culture research and how it impacts on production of biopharmaceutical proteins worldwide, the book is essential reading for everyone working in cell culture and is a recommended volume for all biotechnology libraries.


Protein Expression in Animal Cells

Protein Expression in Animal Cells
Author:
Publisher: Academic Press
Total Pages: 399
Release: 1994-10-18
Genre: Science
ISBN: 0080859410

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Critically acclaimed for more than 25 years, the Methods in Cell Biology series provides an indispensable tool for the researcher. Each volume is carefully edited by experts to contain state-of-the-art reviews and step-by-step protocols. Techniques are described completely so that methods are made accessible to users. Describes both well-established and novel recombinant vector systems for expression of proteins Presents methods for efficient delivery of recombinant genes into differentiated cells, tissues, and whole animals Covers high-level and inducible systems, plus assays for protein expression Provides beginning and advanced investigators and students with the information they need to choose the optimal viral or plasmid system for their protein Practical, benchtop-style presentation works in lab and in the classroom


Cell Line Development

Cell Line Development
Author: Mohamed Al-Rubeai
Publisher: Springer Science & Business Media
Total Pages: 259
Release: 2009-08-11
Genre: Medical
ISBN: 9048122457

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Mammalian cell lines command an effective monopoly for the production of therapeutic proteins that require post-translational modifications. This unique advantage outweighs the costs associated with mammalian cell culture, which are far grater in terms of development time and manufacturing when compared to microbial culture. The development of cell lines has undergone several advances over the years, essentially to meet the requirement to cut the time and costs associated with using such a complex hosts as production platforms. This book provides a comprehensive guide to the methodology involved in the development of cell lines and the cell engineering approach that can be employed to enhance productivity, improve cell function, glycosylation and secretion and control apoptosis. It presents an overall picture of the current topics central to expression engineering including such topics as epigenetics and the use of technologies to overcome positional dependent inactivation, the use of promoter and enhancer sequences for expression of various transgenes, site directed engineering of defined chromosomal sites, and examination of the role of eukaryotic nucleus as the controller of expression of genes that are introduced for production of a desired product. It includes a review of selection methods for high producers and an application developed by a major biopharmaceutical industry to expedite the cell line development process. The potential of cell engineering approch to enhance cell lines through the manipulation of single genes that play important roles in key metabolic and regulatory pathways is also explored throughout.


Gene Transfer and Expression in Mammalian Cells

Gene Transfer and Expression in Mammalian Cells
Author: S.C. Makrides
Publisher: Newnes
Total Pages: 776
Release: 2003-10-24
Genre: Science
ISBN: 0080930352

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The production of proteins in mammalian cells is an important tool in numerous scientific and commercial areas. For example, proteins for human therapy, vaccination or diagnostic applications are typically produced in mammalian cells. Gene cloning, protein engineering, biochemical and biophysical characterization of proteins also require the use of gene expression in mammalian cells. Other applications in widespread use involve screening of libraries of chemical compounds in drug discovery, and the development of cell-based biosensors.This book presents a state-of-the-art comprehensive coverage of the technical aspects of gene expression in mammalian cells, written by experienced scientists working at the forefront of the field.


Protein Expression Technologies

Protein Expression Technologies
Author: François Baneyx
Publisher: Garland Science
Total Pages: 546
Release: 2004
Genre: Medical
ISBN: 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.