植物学技术

Over the last decade, plant-based production of pharmaceuticals has made remarkable progress as the expression of a diverse set of proteins has been demonstrated in a range of plant crops. Although the commercial exploitation is still pending, today various plant-based expression tec ...

Stable accumulation of storage proteins, lipids and carbohydrates is a hallmark of the plant seed, and is a characteristic that is typically deficient in existing platforms for recombinant protein manufacture. One of the biological sequestration mechanisms that facilitate the fo ...

We describe a general approach for the use of recombinant protease inhibitors as stabilizing agents for clinically useful proteins extracted from transgenic plant tissues. A procedure is first described to assess the overall (in)stability of heterologous proteins in transgenic p ...

Transgenic plants are gaining increasing attention from the industry as a natural bioreactor for the production of industrial and chemical products. Optimization of transgene expression in plant cells holds the key to maximizing the potential of plants for producing proteins of com ...

In this chapter, we discuss and compare the different concepts and examples as well as present the basic protocols for applying intrabody-based approaches in plants for the investigation of cell functions and plant cell–pathogen interactions. The immunomodulation strategy, a mole ...

This chapter presents a general procedure for the on-chip detection and quantitation of low-molecular-weight recombinant proteins in transgenic plant crude extracts by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS). A prot ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...

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 ...