Subcellular Targeting and Purification of Recombinant Proteins in Plant Production Systems

Moloney, Maurice M.; Holbrook, Larry A.

doi:10.1080/02648725.1997.10647947

Cited by 26 publications

(8 citation statements)

References 5 publications

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“…In many cases, these peptide sequences can be recognized by a membrane-bound receptor or docking protein, translocated across the membrane along with their host storage proteins, and then trimmed from the storage protein during the process (Chrispeels, 1991;Herman & Larkins, 1999). The targeting determinant peptides of storage protein have been utilized in transgenic plants to target recombinant proteins and enzymes into the designated cellular compartments, such as ER-originated protein body, vacuole-originated protein body, and apoplast (Conrad and Fiedler, 1998;Moloney and Holbrook, 1997). Some approaches have resulted in extremely high-level accumulation of recombinant proteins.…”

Section: Introductionmentioning

confidence: 99%

High yield recombinant silk-like protein production in transgenic plants through protein targeting

et al. 2005

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DP1B is a synthetic analogue of spider dragline silk protein. It can be spun to form silk fiber. Previously, it had been expressed in transgenic plants, showing the general feasibility of the plant-based DP1B production. However, success of such a plant-based platform requires a great increase of DP1B productivity in plant cells to reduce production cost. This report describes a protein targeting approach to accumulate DP1B in apoplast, ER lumen, and vacuole in Arabidopsis cells, by utilizing appropriate combinations of sporamin-targeting determinant peptides and ER retention peptide. The approach has dramatically enhanced DP1B accumulation, resulting in high production yield. The accumulation can be as high as 8.5 and 6.7% total soluble protein in leaf tissue by targeting to apoplast and ER lumen, respectively, or as high as 18 and 8.2% total soluble protein in seeds by targeting to ER lumen and vacuole, respectively. However, the vacuole targeting in leaves and the apoplast targeting in seeds have failed to accumulate full length DP1B molecules or any DP1B at all, respectively, suggesting that they may not be suitable for applications in leaf tissues and seeds. Data in this study recommend a combination of seed-specific expression and ER-targeting as one of the best strategies for yield enhancement of plant-based DP1B production.

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Section: Introductionmentioning

confidence: 99%

High yield recombinant silk-like protein production in transgenic plants through protein targeting

et al. 2005

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“…glycosylation) and this can subsequently induce adverse immunogenic reactions in patients when they enter to the circulatory system [ 62 ]. Protein glycosylation, particularly N -glycosylation, initiates in the ER and continues in the endomembrane system of the secretory pathway by the function of many glycosidases and glycosyltransferases [ 54 , 59 , 63 ].…”

Section: Genetic Strategies For the Manipulation Of Subcellular Targementioning

confidence: 99%

Theoretical Basis of Plant Cell and Tissue Culture for Production of Biomass and Bioactive Compounds

López‐Villalobos

Yeung

Thorpe

2014

Production of Biomass and Bioactive Compounds Using Bioreactor Technology

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Plant tissue culture is an important biotechnological tool, which involves biochemical and genetic manipulations to onset specifi c gene programming, which determines an optimal cell differentiation state for the production of bioactive compounds. Indeed, not only the metabolism but also the morphogenetic processes are modifi ed in plant cells to drive the synthesis and accumulation of economically valuable compounds. In this chapter, we provide an overview of the current molecular approaches to control the expression of specifi c genes encoding putative heterologous and native enzymes as well as transcription factors to enhance the metabolic fl ow of specifi c pathways in order that notable bioactive compounds can be accumulated in plant cells at acceptable commercial levels. Such methods vary from singlegene plant transformations to the emerging multi-gene transformation (gene stacking) technologies embraced by private companies focusing primarily in the metabolic engineering of secondary metabolism. The virtues and potential of these molecular methods in their application to tissue culture systems are presented. Additionally, the importance of subcellular targeting of proteins, notably biosynthetic enzymes and pharmaceutical antibodies, for enhancing their activity and stability is also discussed. Finally, the progress in two emerging approaches for the production of bioactive molecules, the manipulation of cell differentiation and cell immortalization are expounded in this article. Thus, we present the molecular basis to control both cell differentiation and cell immortalization in plant tissue culture systems as novel avenues to control and perpetuate the gene programming which in turn creates and regulates cellular microenvironments for the optimal biosynthesis of valuable compounds. Consequently, our objective is to present how basic approaches, including the manipulation of gene expression, are amalgamated to other molecular strategies of higher hierarchy, particularly the manipulation of cell differentiation and immortalization for the synthesis of bioactive molecules in plant tissue culture platforms.

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“…This depends on the nature of any targeting signals on the expression construct (see above) and the permeability of the plant cell wall to macromolecules [88]. Targeting signals can be used to direct the protein to the secretory pathway [89] or to intracellular organelles such as the chloroplast or vacuole [90]. As shown for transgenic plants, the cytosol is generally unsuitable for the accumulation of recombinant antibodies because incorrect folding and assembly lead to rapid degradation.…”

Section: Plant Suspension Cells For Protein Productionmentioning

confidence: 99%

“…Protocols for the recovery of recombinant antibodies from animal cells, serum and microbial cells are refined and well described. However, there are few reports of purification methods for recombinant antibodies from plant suspension cells, leaves or seeds [90]. In this context, the development of transgenic plants that secrete recombinant proteins could be useful.…”

Section: Downstream Processing Of Recombinant Antibodies From Plant Cmentioning

confidence: 99%

Molecular farming of recombinant antibodies in plants

Schillberg

Fischer

Emans³

2003

Cellular and Molecular Life Sciences (CMLS)

142

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Antibodies represent a large proportion of therapeutic drugs currently in development. In most cases, they are produced in mammalian cell lines or transgenic animals because these have been shown to fold and assemble the proteins correctly and generate authentic glycosylation patterns. However, such expression systems are expensive, difficult to scale up and there are safety concerns due to potential contamination with pathogenic organisms or oncogenic DNA sequences. Plants represent an inexpensive, efficient and safe alternative for the production of recombinant antibodies. Research over the last 10 years has shown that plants can produce a variety of functional antibodies and there is now intense interest in scaling up production to commercial levels. In this review, we discuss the advantages of plants over traditional expression systems, describe how antibody expression in plants is achieved and optimized and then consider the practical issues concerning large-scale molecular farming in plants. The first plant-produced therapeutic antibodies are already in clinical trials, and, given the economic benefits of this production system, we are likely to see many more recombinant antibodies produced in this manner in the future.

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Subcellular Targeting and Purification of Recombinant Proteins in Plant Production Systems

Cited by 26 publications

References 5 publications

High yield recombinant silk-like protein production in transgenic plants through protein targeting

High yield recombinant silk-like protein production in transgenic plants through protein targeting

Theoretical Basis of Plant Cell and Tissue Culture for Production of Biomass and Bioactive Compounds

Molecular farming of recombinant antibodies in plants

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