Animal Cell Biotechnology: Methods and Protocols (Methods in Biotechnology)
Mammalian cells are used in industry as well as in research for a variety of applications. Examples are the production of monoclonal antibodies with hybridoma cells or proteins for diagnostic or therapeutic use with recombinant cells, production of viral vaccines, as well as cultivation of tissue cells for artificial organs or for gene therapy. Beside the techniques required for establishing specific cell lines are techniques required for optimized cultivation in small and large scale cell characterization and analysis and purification of biopharmaceuticals and vaccines.
Animal Cell Biotechnology: Methods and Protocols is divided into parts that reflect the processes required for different stages of production. In Part I basic techniques for establishment of production cell lines are addressed. Safe and reliable transfer of genetic information at high frequencies into desired target cells remains one of the major challenges in therapeutic life sciences. Multiply attenuated viral vector systems, which are able to transfer heterologous DNA or RNA into almost any cell phenotype as well as in vivo, emerged as the most efficient transgene delivery systems in modern biology.
The treatment of genetic diseases and of several acquired diseases can only reasonably be performed by using gene or cell therapy approaches. Chapter 1 provides a comprehensive technical overview of the most commonly used viral transduction systems and compares their specific characteristics in order for researchers to make the best choice in a particular scientific setting. In Chapter 2, in addition to protocols for the establishment of producer cells and the production of viral vectors, some basic titration methods are presented.
Monoclonal antibodies (mAbs) are unique and versatile molecules that have been applied in research, as well as to in vitro and in vivo diagnostics and treatment of several diseases. Chapter 3 is an overview of the state of the art of antibody engineering and production methods and describes standard protocols for producing a hybridoma cell line the building up of transfectomas. A suitable bacterial expression system for the production of antibodies and antibody fragments is also given.
The history of the culture of animal cell lines is littered with published and much unpublished experience with cell lines that have become switched, mislabelled, or cross-contaminated during laboratory handling. To deliver valid and good quality research and to avoid expenditure of time and resources on such rogue lines, it is vital to perform some kind of qualification for the provenance of cell lines used in research and particularly in the development of biomedical products. DNA fingerprinting provides a valuable tool to compare different sources of the same cells and, where original material or tissue is available, to confirm the correct identity of a cell line. Chapter 4 provides a review of some of the most useful techniques to test the identity of cells in the cell culture laboratory and presents methods used in the authentication of cell lines.
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|March 24, 2017|
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