Mohamed Al-Rubeai – författare

This work present practical, biotechnological applications of flow cytometry techniques for the study of animal, plant and microbial cells, explaining methodologies for sample preparation, staining and analysis. It discusses cell variability in cell culture processes and shows how the quantitative analysis of heterogeneous populations aids in the biotechnological exploitation of cells.

Integrating advances in molecular biology into bioprocesses presents a continuous challenge to scientists and bioengineers. This series is conceived to help meet this challenge. The book examines and assesses the feasibility of new approaches for the modification of cellular function such as gene expression, protein processing, secretion, glycosylation, immortalisation, proliferation, and apoptosis as well as the systematic study of the metabolic genotype-phenotype relationship. The series provides detailed coverage of the methodology for improving cellular properties of cells used in the production of biopharmaceuticals, gene and cell therapies and tissue engineering. It also seeks to explain the cellular mechanisms underlying in vitro physiological activity and productivity. This volume, which is based on presentations at the "European Workshop on Animal Cell Engineering" held in Costa Brava, Spain, contains a collection of chapters relating to cellular function and modification by leading authorities in several different areas of basic research and the biopharmaceutical industry.

This volume focuses on the transient gene expression technologies for recombinant protein production and gene therapy. Topics covered range from the use of baculovirus, adenovirus, adeno-associated virus and alphavirus vectors in biopharmaceutical, genomics and gene delivery research, to large-scale transfection of plasmid DNA into cultured cells for bioreactor applications. The book illuminates all the scientific and technological issues involved in the research, development, production, and human use of viral and non-viral vectors for in vitro and in vivo expression of recombinant proteins, thus making it an useful reference for cell culture technologists, virologists, and scientists interested in gene therapy.

This work present practical, biotechnological applications of flow cytometry techniques for the study of animal, plant and microbial cells, explaining methodologies for sample preparation, staining and analysis. It discusses cell variability in cell culture processes and shows how the quantitative analysis of heterogeneous populations aids in the biotechnological exploitation of cells.

The suppression of apoptosis by the IGF system is critical for normal cell development, proliferation, differentiation and motility. Aberrations in IGF signalling mechanisms contribute to cell transformation, tumour progression and metastasis. Many questions remain to be answered as to how exactly the IGF system mediates its effects both in normal and tumour cells and how the IGF-1R interacting proteins and downstream signalling cascades are regulated. The importance of the IGF system is underscored by the significant interest in the development of anti-IGF therapies for IGF sensitive cancers. Future developments in cancer therapy are likely to focus on methods to target these therapies to diseased but not normal cells. 14. Acknowledgements We would like to thank Kurt Tidmore for preparing the illustrations. The Health Research Board of Ireland and Science Foundation Ireland are grateful acknowledged for funding. 15. References Adamo M., Roberts C. T., Jr. and LeRoith D. (1992) How distinct are the insulin and insul- like growth factor I signalling systems? Biofactors 3, 151-7. Adams T. E., Epa V. C., Garrett T. P. and Ward C. W.(2000) Structure and function of the type 1 insulin-like growth factor receptor. Cell Mol Life Sci 57, 1050-93. Adler V., Polotskaya A., Wagner F. and Kraft A. S. (1992) Affinity-purified c-Jun ami- terminal protein kinase requires serine/threonine phosphorylation for activity. J Biol Chem 267, 17001-5.

For the first time in a single volume, the design, characterisation and operation of the bioreactor system in which the tissue is grown is detailed.Bioreactors for Tissue Engineering presents an overall picture of the current state of knowledge in the engineering of bioreactors for several tissue types (bone, cartilage, vascular), addresses the issue of mechanical conditioning of the tissue, and describes the use of techniques such as MRI for monitoring tissue growth.This unique volume is dedicated to the fundamentals and application of bioreactor technology to tissue engineering products. Not only will it appeal to graduate students and experienced researchers in tissue engineering and regenerative medicine, but also to tissue engineers and culture technologists, academic and industrial chemical engineers, biochemical engineers and cell biologists who wish to understand the criteria used to design and develop novel systems for tissue growth in vitro.

A comprehensive guide to the revolutionary area of systems biology and its application in cell culture engineering, this volume presents an overall picture of the current topics central to structural and functional genomics, proteomics, metabolomics and bioinformatics, including such hot topics as RNAi, metabolic engineering and unfolded protein response. It includes reviews of the cellular response of environmental modulation such as low temperature and osmolarity, critical assessments of the applications of metabolomics and fluxomics approaches, examination of the utility of modulation of key genes and a presentation of a theory of chemical organisation which provides a new view of the system's structure. The clearly written chapters by experts in the field describe methods applicable to investigating the unique facets of cell culture. The book should be of interest to all those working in cell culture development and drug discovery in pharmaceutical and biotechnology companies as well as in academic institutions. It provides an invaluable resource for students and researchers in biotechnology, cell culture, genomics and bioinformatics.

The huge potential for gene therapy to cure a wide range of diseases has led to high expectations and a great increase in research efforts in this area, particularly in the study of delivery via viral vectors, widely considered to be more efficient than DNA transfection.  In Viral Vectors for Gene Therapy: Methods and Protocols, experts in the field present a collection of their knowledge and experience featuring methodologies that involve virus production, transferring protocols, and evaluating the efficacy of gene products. While thoroughly covering the most popular viral vector systems of adenovirus, retrovirus, and adeno-associated virus, this detailed volume also explores less common viral vector systems such as baculovirus, herpes virus, and measles virus, the growing interest in which is creating a considerable demand for large scale manufacturing and purification procedures.  Written in the highly successful Methods in Molecular Biology™ series format, many chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Viral Vectors for Gene Therapy: Methods and Protocols provides basic principles accessible to scientists from a wide variety of backgrounds for the development of gene therapy viral products that are safe and effective.

The huge potential for gene therapy to cure a wide range of diseases has led to high expectations and a great increase in research efforts in this area, particularly in the study of delivery via viral vectors, widely considered to be more efficient than DNA transfection.  In Viral Vectors for Gene Therapy: Methods and Protocols, experts in the field present a collection of their knowledge and experience featuring methodologies that involve virus production, transferring protocols, and evaluating the efficacy of gene products. While thoroughly covering the most popular viral vector systems of adenovirus, retrovirus, and adeno-associated virus, this detailed volume also explores less common viral vector systems such as baculovirus, herpes virus, and measles virus, the growing interest in which is creating a considerable demand for large scale manufacturing and purification procedures.  Written in the highly successful Methods in Molecular Biology™ series format, many chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and vital tips on troubleshooting and avoiding known pitfalls. Comprehensive and practical, Viral Vectors for Gene Therapy: Methods and Protocols provides basic principles accessible to scientists from a wide variety of backgrounds for the development of gene therapy viral products that are safe and effective.

Microbial fermentation is a powerful tool with a bright future, but it also faces some challenges. This contributed volume is dedicated toward some of the current and contemporary issues in this field, such as:Scaling up production: Many exciting products can be created through fermentation, but scaling these processes from the lab to industrial production can be difficult. This involves optimising conditions for large-scale fermentation tanks, ensuring consistent product quality, and keeping costs competitive.Strain development: Finding or engineering the most efficient and productive microbial strains is crucial. Researchers are looking for ways to improve the yield and desired product formation of these microbes. This might involve techniques like genetic modification or harnessing the vast diversity of the microbial world to find new players.Sustainability: Fermentation processes need to be environmentally friendly. Finding sustainable feedstocks (the starting materials for fermentation) is important, especially as we move away from reliance on fossil fuels. Additionally, minimizing waste products and energy consumption during fermentation are key areas of focus.Integration with other technologies: The future of fermentation is likely to involve its integration with other technologies. For instance, combining fermentation with advanced materials science or artificial intelligence could lead to even more innovative applications.By addressing these issues, researchers and industry leaders can unlock the full potential of microbial fermentation and create a more sustainable and innovative future.

​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.

​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.

The aim of this volume is to provide an in-depth overview of the state-of-the-art research on apoptosis with contributions from key groups working in the field. This type of programme cell death has received wide and rapid attention and now is considered as one of the hottest areas of science. The volume covers various aspects of the apoptotic death process from the morphological and biochemical features, mechanisms and genetic regulation to its role in pathological process and potential implications for biomedical research and biopharmaceutical production.

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.

Integrating advances in molecular biology into bioprocesses presents a continuous challenge to scientists and bioengineers. This series is conceived to help meet this challenge. It examines and assesses the feasibility of new approaches for the modification of cellular function such as gene expression, protein processing, secretion, glycosylation, immortalisation, proliferation, and apoptosis as well as the systematic study of the metabolic genotype-phenotype relationship. The series provides detailed coverage of the methodology for improving cellular properties of cells used in the production of biopharmaceuticals, gene and cell therapies and tissue engineering. It also seeks to explain the cellular mechanisms underlying in vitro physiological activity and productivity. This volume, which is based on presentations at the 'European Workshop on Animal Cell Engineering' held in Costa Brava, Spain, contains a collection of chapters relating to cellular function and modification by leading authorities in several different areas of basic research and the biopharmaceutical industry.

The suppression of apoptosis by the IGF system is critical for normal cell development, proliferation, differentiation and motility. Aberrations in IGF signalling mechanisms contribute to cell transformation, tumour progression and metastasis. Many questions remain to be answered as to how exactly the IGF system mediates its effects both in normal and tumour cells and how the IGF-1R interacting proteins and downstream signalling cascades are regulated. The importance of the IGF system is underscored by the significant interest in the development of anti-IGF therapies for IGF sensitive cancers. Future developments in cancer therapy are likely to focus on methods to target these therapies to diseased but not normal cells. 14. Acknowledgements We would like to thank Kurt Tidmore for preparing the illustrations. The Health Research Board of Ireland and Science Foundation Ireland are grateful acknowledged for funding. 15. References Adamo M., Roberts C. T., Jr. and LeRoith D. (1992) How distinct are the insulin and insul- like growth factor I signalling systems? Biofactors 3, 151-7. Adams T. E., Epa V. C., Garrett T. P. and Ward C. W.(2000) Structure and function of the type 1 insulin-like growth factor receptor. Cell Mol Life Sci 57, 1050-93. Adler V., Polotskaya A., Wagner F. and Kraft A. S. (1992) Affinity-purified c-Jun ami- terminal protein kinase requires serine/threonine phosphorylation for activity. J Biol Chem 267, 17001-5.

For the first time in a single volume, the design, characterisation and operation of the bioreactor system in which the tissue is grown is detailed.Bioreactors for Tissue Engineering presents an overall picture of the current state of knowledge in the engineering of bioreactors for several tissue types (bone, cartilage, vascular), addresses the issue of mechanical conditioning of the tissue, and describes the use of techniques such as MRI for monitoring tissue growth.This unique volume is dedicated to the fundamentals and application of bioreactor technology to tissue engineering products. Not only will it appeal to graduate students and experienced researchers in tissue engineering and regenerative medicine, but also to tissue engineers and culture technologists, academic and industrial chemical engineers, biochemical engineers and cell biologists who wish to understand the criteria used to design and develop novel systems for tissue growth in vitro.

A comprehensive guide to the revolutionary area of systems biology and its application in cell culture engineering, this volume presents an overall picture of the current topics central to structural and functional genomics, proteomics, metabolomics and bioinformatics, including such hot topics as RNAi, metabolic engineering and unfolded protein response. It includes reviews of the cellular response of environmental modulation such as low temperature and osmolarity, critical assessments of the applications of metabolomics and fluxomics approaches, examination of the utility of modulation of key genes and a presentation of a theory of chemical organisation which provides a new view of the system's structure. The clearly written chapters by experts in the field describe methods applicable to investigating the unique facets of cell culture. The book should be of interest to all those working in cell culture development and drug discovery in pharmaceutical and biotechnology companies as well as in academic institutions. It provides an invaluable resource for students and researchers in biotechnology, cell culture, genomics and bioinformatics.

Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.

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.

Engineered antibodies currently represent over 30% of biopharmaceuticals in clinical trials and their total worldwide sales continue to increase significantly. The importance of antibody applications is reflected in their increasing clinical and industrial applications as well as in the progression of established and emerging production strategies. This volume provides detailed coverage of the generation, optimization, characterization, production and applications of antibody. It provides the necessary theoretical background and description of methods for the expression of antibody in microbial and animal cell cultures and in transgenic animals and plants. There is a strong focus on those issues related to the production of intrabodies, bispecific antibody and antibody fragments and also to novel applications in cancer immunotherapy.

With the discovery of stem cells capable of multiplying indefinitely in culture and differentiating into many other cell types in appropriate conditions, new hopes were born in repair and replacement of damaged cells and tissues. The features of stem cells may provide treatment for some incurable diseases with some therapies are already in clinics, particularly those from adult stem cells. Some treatments will require large number of cells and may also require multiple doses, generating a growing demand for generating and processing large numbers of cells to meet the need of clinical applications. With this in mind, our aim is to provide a book on the subject of stem cells and cell therapy for researchers and students of cell biotechnology, bioengineering and bioproduction. This book is exceptional as it teaches researchers stem cells and cell therapy in that it covers the concepts and backgrounds necessary so that readers get a good understanding of the production of stem cells. The book covers three topics: The basics of stem cells and cell therapy, the use of stem cells for the treatment of human diseases, and stem cell processing. It includes chapters on neural and vascular stem vascular stem cell therapy, expansion engineering of embryonic stem cells, stem cell based production of blood cells and separation technologies for stem cells and cell therapy products. It is an informed and informative presentation of what modern research, science and engineering have learned about stem cells and their production and therapies. Addressing both the medical and production issues, this book is an invaluable contribution to having an academic and industrial understanding with respect to R&D and manufacturing of clinical grade stem cells.

The advantages of the baculovirus system are rooted in the properties of the virus and the host (insect, or cell lines derived from it). During the normal infection cycle, two forms of the virus are produced: an early budded virus (BY) form (Kost et al. , 2000), in which the viral DNA and structural proteins are surrounded by membrane derived from the infected cell; and a late occluded form (occlusion-derived virus, ODy), consisting of enveloped viral cores which are embedded in a crystal matrix of viral proteins. The principal component of the matrix is the abundantly expressed protein polyhedrin. The budded virus rapidly spreads the infection from cell to cell within the insect host, resulting ultimately in the complete liquefaction of the host, and release of occluded virus into the environment. The occluded form protects the released virus, allowing it to survive for long periods in the environment until ingested by another host. In the alkaline environment ofthe insect gut, the protective protein matrix is removed, and the life cycle is repeated. In insect cell cultures, only the BV form of baculovirus is required, and the polyhedrin gene may be replaced with the gene for the recombinant protein. An additional benefit of replacing or deleting polyhedrin is that it effectively makes the virus unable to survive outside the laboratory, an advantage in terms of environmental safety. The system is intrinsically safe to animals, being unable to replicate in species other than a limited range of insects.