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From Neanderthal to Nanobiotech: From Plant Potions to Pharming with Plant Factories

Plants were the main source for human drugs until the beginning of the nineteenth century when plantderived pharmaceuticals were partly supplanted by drugs produced by the industrial methods of chemical synthesis. During the last decades of the twentieth century, genetic engineeri ...

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Assessing the Risk of Undesirable Immunogenicity/ Allergenicity of Plant-Derived Therapeutic Proteins

Therapeutic proteins have an intrinsic potential to induce undesirable immune and allergic responses. The nature of the expression system and the control of the manufacturing process represent extrinsic factors that could modify this potential. Accordingly, regulatory agen ...

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Biosafety, Risk Assessment and Regulation of Plant-Made Pharmaceuticals

The technology for plant-made pharmaceuticals (PMPs) has progressed significantly over the last few years, with the first commercial products for human use expected to reach the market by 2009 (see Note 1 ). As part of the ‘next generation’ of genetically modified (GM) crops, PMPs will be subject to ...

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Cowpea Mosaic Virus-Based Systems for the Expression of Antigens and Antibodies in Plants

This chapter describes the use of Cowpea mosaic virus-based vectors for the production of foreign proteins such as antigens and antibodies in plants. The systems include vectors based on both full-length and deleted versions of RNA-2. In both cases, the modified RNA-2 is replicated by coinocu ...

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Transient Expression of Antibodies in Plants Using Syringe Agroinfiltration

The improvements in agroinfiltration methods for plant-based transient expression now allow the production of significant amounts of recombinant proteins in a matter of days. While vacuum-based agroinfiltration has been brought to large scale to meet the cost, speed and surge capac ...

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Rapid System for Evaluating Bioproduction Capacity of Complex Pharmaceutical Proteins in Plants

Transgene product yield remains a key limitation in commercializing plant-derived pharmaceutical proteins. Although significant progress has been made in understanding the roles of promoters, enhancers, integration sites, codon usage, cryptic RNA sites, silencing, and pro ...

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Production and Localization of Recombinant Pharmaceuticals in Transgenic Seeds

Among the many plant-based production systems that have been developed for pharmaceutical proteins, seeds have the useful advantage of accumulating proteins in a relatively small volume, and recom-binant proteins are very stable in dry seeds allowing long-term storage and facilit ...

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Production of Antibody Fragments in Arabidopsis Seeds

Plants offer a number of attractive benefits over conventional mammalian or bacterial cell culture systems for the production of valuable pharmaceutical and industrial proteins. Currently, antibodies and their derived fragments represent the largest and most important group ...

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Production of Plantibodies in Nicotiana Plants

Because of the wide use and high demand in medicine, monoclonal antibodies are among the main recombinant pharmaceuticals at present, although present limitations of the productive platforms for monoclonal antibodies are driving the improvement of the large-scale technologies ...

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Physcomitrella patens : A Non-Vascular Plant for Recombinant Protein Production

The moss Physcomitrella patens is a long-standing model for studying plant development, growth and cell differentiation in particular. Interest in this non-vascular plant arose following the discovery that homologous recombination is an efficient process. P. patens is, therefo ...

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Production of Recombinant Proteins in SuspensionCultured Plant Cells

Plants have emerged in the past decade as a suitable alternative to the current production systems for recombinant pharmaceutical proteins and, today their potential for low-cost production of high quality, much safer and biologically active mammalian proteins is largely documen ...

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Isolation of Cell Wall Proteins from Medicago sativa Stems

Plant cell walls are highly dynamic and chemically active components of plant cells. Cell walls consist primarily of polysaccharides, with proteins comprising approx 10% of the cell wall mass. These proteins are difficult to isolate with a high degree of purity from the complex carbohydrate ...

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Two-Dimensional Differential In-Gel Electrophoresis (DIGE) of Leaf and Roots of Lycopersicon esculentum

In this report we present a detailed protocol for the analysis of differential protein expression between two plant tissue samples. The protocol involves harvesting of leaves and roots from mature tomato plants, preparing protein extracts from the harvested tissues, fluorescent la ...

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Visible and Fluorescent Staining of Two-Dimensional Gels

Staining of two-dimensional gel constitutes a crucial step in comparative proteome analysis with respect to the number of proteins analyzed, the accuracy of spot quantification, and compatibility with mass spectrometry. The present chapter describes procedures for several vis ...

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Two-Dimensional Electrophoresis for Plant Proteomics

Two-dimensional gel electrophoresis (2-DE) with immobilized pH gradients (IPGs) combined with protein identification by mass spectrometry (MS) is currently the workhorse for proteome analysis. 2-DE allows separation of highly complex mixtures of proteins according to isoel ...

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Detergents and Chaotropes for Protein Solubilization before Two-Dimensional Electrophoresis

Because of the outstanding separating capabilities of two-dimensional electrophoresis for complete proteins, it would be advantageous to be able to apply it to all types of proteins. Unfortunately, severe solubility problems hamper the analysis of many classes of proteins, but espe ...

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Plant Plasma Membrane Protein Extraction and Solubilization for Proteomic Analysis

The plasma membrane (PM) exists as the interface between the cytosol and the environment in all living cells and is one of the most complex and differentiated membrane. The identification and characterization of membrane proteins (either extrinsic or intrinsic) is a crucial challenge si ...

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Quantitative Analysis of 2D Gels

The use of 2D electrophoresis for comparative proteomics allows the revelation of variations of protein relative amounts according to various physiological or genetic criteria. The statistical significance of these results is related to different factors, from the experiment ...

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Extraction of Nuclear Proteins

The integrity of a subcellular proteome such as the nucleus, is largely dependent on purification of the isolated compartment away from other cellular contaminants. The separation of high-purity nuclei from plants is a difficult task. However, successful purification has been achie ...

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Extraction of Nuclear Proteins from Root Meristematic Cells

Fractionation and extraction of nuclear proteins are techniques intended to facilitate dedicated plant proteomic studies. These techniques rely on subcellular fractionation, which makes it possible to define and characterize the proteome of a subcellular organelle, in this c ...

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