Recombinant Proteins From Plants - Methods and Protocols

Altogether, the biochemical, technical and economic limitations on existing proka- otic and eukaryotic expression systems and the growing clinical demand for complex therapeutic proteins have created substantial interest in developing new expression systems for the production of therapeutic proteins. To that end, plants have emerged in the past decade as a suitable alternative to the current production systems, and today their potential for production of high quality, much safer and biologically active complex recombinant pharmaceutical proteins is largely documented. The chapters in this volume, contributed by leaders in the field, sum up the state-- the-art methods for using a variety of different plants as expression hosts for phar- ceutical proteins. Several production platforms are presented, ranging from seed- and leaf-based production in stable transgenic plant lines, to plant cell bioreactors, to viral or Agrobacterium-mediated transient expr ession systems. Currently, antibodies and their derived fragments represent the largest and most important group of biote- nological products in clinical trials. This explains why the potential of most prod- tion platforms is illustrated here principally for antibodies or antibody fragments with acknowledged potential for immunotherapy in humans. In addition, a comparison of different plant expression systems is presented using aprotinin, a commercial phar- ceutical protein, as a test system. Although multiple books and monographs have been recently published on mol- ular pharming, there is a noticeable dearth of bench step-by-step protocols that can be used quickly and easily by beginners entering this new field.

List of contents

From Neanderthal to Nanobiotech: From Plant Potions to Pharming with Plant Factories.- Cowpea Mosaic Virus-Based Systems for the Expression of Antigens and Antibodies in Plants.- Transient Expression of Antibodies in Plants Using Syringe Agroinfiltration.- Rapid System for Evaluating Bioproduction Capacity of Complex Pharmaceutical Proteins in Plants.- Production and Localization of Recombinant Pharmaceuticals in Transgenic Seeds.- Production of Antibody Fragments in Arabidopsis Seeds.- Production of Plantibodies in Nicotiana Plants.- Physcomitrella patens : A Non-Vascular Plant for Recombinant Protein Production.- Production of Recombinant Proteins in Suspension-Cultured Plant Cells.- Chloroplast-Derived Vaccine Antigens and Biopharmaceuticals: Protocols for Expression, Purification, or Oral Delivery and Functional Evaluation.- Protein Body Induction: A New Tool to Produce and Recover Recombinant Proteins in Plants.- A Case Study for Plant-Made Pharmaceuticals Comparing Different Plant Expression and Production Systems.- Glycosylation of Antibody Therapeutics: Optimisation for Purpose.- N-Glycosylation of Plant Recombinant Pharmaceuticals.- Companion Protease Inhibitors to Protect Recombinant Proteins in Transgenic Plant Extracts.- Strategies for Improving Vaccine Antigens Expression in Transgenic Plants: Fusion to Carrier Sequences.- Immunomodulation of Plant Function by In Vitro Selected Single-Chain Fv Intrabodies.- On-Chip Detection of Low-Molecular-Weight Recombinant Proteins in Plant Crude Extracts by SELDI-TOF MS.- Assessing the Risk of Undesirable Immunogenicity/ Allergenicity of Plant-Derived Therapeutic Proteins.- Biosafety, Risk Assessment and Regulation of Plant-Made Pharmaceuticals.

Summary

Altogether, the biochemical, technical and economic limitations on existing proka- otic and eukaryotic expression systems and the growing clinical demand for complex therapeutic proteins have created substantial interest in developing new expression systems for the production of therapeutic proteins. To that end, plants have emerged in the past decade as a suitable alternative to the current production systems, and today their potential for production of high quality, much safer and biologically active complex recombinant pharmaceutical proteins is largely documented. The chapters in this volume, contributed by leaders in the field, sum up the state-- the-art methods for using a variety of different plants as expression hosts for phar- ceutical proteins. Several production platforms are presented, ranging from seed- and leaf-based production in stable transgenic plant lines, to plant cell bioreactors, to viral or Agrobacterium-mediated transient expr ession systems. Currently, antibodies and their derived fragments represent the largest and most important group of biote- nological products in clinical trials. This explains why the potential of most prod- tion platforms is illustrated here principally for antibodies or antibody fragments with acknowledged potential for immunotherapy in humans. In addition, a comparison of different plant expression systems is presented using aprotinin, a commercial phar- ceutical protein, as a test system. Although multiple books and monographs have been recently published on mol- ular pharming, there is a noticeable dearth of bench step-by-step protocols that can be used quickly and easily by beginners entering this new field.

Additional text

From the reviews:

“This book presents current methods for the expression of those pharmaceutical proteins in a variety of plant systems. … Each one of its twenty chapters offers objective, clearly written texts including a summary … and some important notes followed by the references. … ‘Recombinant proteins from plants’ is surely a useful tool and a practical and rapid summarized data source with easy procedures on plant molecular pharming for all scientists and related professionals working in this growing field.” (Dominique C. H. Fischer, Brazilian Journal of Pharmaceutical Sciences, 2009)

Report

From the reviews: "This book presents current methods for the expression of those pharmaceutical proteins in a variety of plant systems. ... Each one of its twenty chapters offers objective, clearly written texts including a summary ... and some important notes followed by the references. ... 'Recombinant proteins from plants' is surely a useful tool and a practical and rapid summarized data source with easy procedures on plant molecular pharming for all scientists and related professionals working in this growing field." (Dominique C. H. Fischer, Brazilian Journal of Pharmaceutical Sciences, 2009)

Product details

• Assisted by: Loï Faye (Editor), Loïc Faye (Editor), Gomord (Editor), Gomord (Editor), Veronique Gomord (Editor)
• Publisher: Springer, Berlin
• Languages: English
• Product format: Paperback / Softback
• Released: 10.12.2010
• EAN: 9781617378645
• ISBN: 978-1-61737-864-5
• No. of pages: 358
• Weight: 937 g
• Illustrations: XVII, 358 p.
• Series: Methods in Molecular Biology; Methods in Molecular Biology
• Subjects: Natural sciences, medicine, IT, technology > Biology > Genetics, genetic engineering; C, Life Sciences, Botany, biochemistry, biotechnology, Proteomics, Biomedical and Life Sciences, Botany & plant sciences, Plant Science, Plant Sciences