Manufacturing of Pharmaceutical Proteins: From Technology to Economy

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Breeding 9 , — This paper considers in detail the factors that should be evaluated when choosing a crop system for the production of pharmaceutical proteins. The same scFv is expressed in many species to compare intrinsic yields, and features such as storage, distribution and biosafety are discussed, as well as economic factors. Witcher, D. Monoclonal antibody manufacturing in transgenic plants myths and realities. From green plants to industrial enzymes. Enzyme Microbial Technol. Zeitlin, L. A humanized monoclonal antibody produced in transgenic plants for immunoprotection of the vagina against genital herpes.

Khoudi, H. Production of a diagnostic monoclonal antibody in perennial alfalfa plants. Perrin, Y. Transgenic pea seeds as bioreactors for the production of a single-chain Fv fragment scFV antibody used in cancer diagnosis and therapy. Breeding 6 , — De Wilde, C. Expression of antibodies and Fab fragments in transgenic potato plants: a case study for bulk production in crop plants.

Plants: The Future Pharmaceutical Factory

Schunmann, P. McGarvey, P. Expression of the rabies virus glycoprotein in transgenic tomatoes. Biotechnology 13 , — Sala, F. Vaccine antigen production in transgenic plants: strategies, gene constructs and perspectives. Vaccine 21 , — Commandeur, U. The biosafety of molecular farming in plants. Hare, P. Excision of selectable marker genes from transgenic plants. Zuo, J. Marker-free transformation: increasing transformation frequency by the use of regeneration-promoting genes. Eastham, K. Environment Issue Report No. Kay, E. In situ transfer of antibiotic resistance genes from transgenic transplastomic tobacco plants to bacteria.

Smalla, K. Extension Press, Univ. Smyth, S. Product differentiation alternatives: identity preservation, segregation and traceability. AgBioForum 5 , 30—42 Molecular farming of recombinant antibodies in plants. Life Sci. This is a comprehensive discussion of the technical issues concerning the production of antibodies in plants, which is treated in much more detail than is possible in the present review.

Plantibodies: applications, advantages and bottlenecks. McCormick, A. Rapid production of specific vaccines for lymphoma by expression of the tumor-derived single-chain Fv epitopes in tobacco plants. USA 96 , — Larrick, J. Production of secretory IgA antibodies in plants. Biomolecular Eng. This paper presents a useful summary of recent advances in the plant-based production of secretory IgAs with a discussion of purification methods and production costs.

Characterization of a recombinant plant monoclonal secretory antibody and preventive immunotherapy in humans. Vaquero, C. A carcinoembryonic antigen-specific diabody produced in tobacco. Kathuria, S. Efficacy of plant-produced recombinant antibodies against HCG. Human Reproduction 17 , — Miele, L. Plants as bioreactors for pharmaceuticals: regulatory considerations. Trends Biotechnol. Emlay, D. Lloyd-Evans, M. Theta Report No. Antibody Engineering: Protocols and Methods 2nd edn ed. Lo, B. Humana Press Inc. Plasma membrane display of anti-viral single chain Fv fragments confers resistance to tobacco mosaic virus.

Sijmons, P. Production of correctly processed human serum albumin in transgenic plants. Biotechnology 8 , — Zhu, Z. Virology , — Matsumoto, S. Characterisation of a human glycoprotein erythropoetin produced in cultured tobacco cells. Delaney, D. Vasil, I. Torres, E.

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Rice cell culture as an alternative production system for functional diagnostic and therapeutic antibodies. Terashima, M. Synthesis and self-assembly of a functional monoclonal antibody in transgenic Nicotiana tabacum. Francisco, J. Expression and characterization of bryodin 1 and a bryodin 1-based single-chain immunotoxin from tobacco cell culture.

ISBN 13: 9783527337668

Mayfield, S. Expression and assembly of a fully active antibody in algae. USA , — Streatfield, S. Plant-based vaccines: unique advantages. Vaccine 19 , — Ma, S. Transgenic plants expressing autoantigens fed to mice to induce oral immune tolerance. Yu, J.

A plant-based multicomponent vaccine protects mice from enteric diseases. Lamphear, B. Delivery of subunit vaccines in maize seed. Control Release 83 , — Download references. The authors are grateful to R. Twyman for critical assessment and help with manuscript preparation.

Correspondence to Julian K-C. Monoclonal antibody derivatives that comprise a single polypeptide in which the variable regions of the heavy and light immunoglobulin chains are joined together by a flexible linker. The large-scale production of recombinant proteins in living cells or organisms; frequently applied to the use of crop plants or domestic animals as expression hosts because of the allusion to agriculture. In the context of this article, a gene or protein that is not derived from the species in which it is expressed.

A transgenic plant in which the transgene is found in the plastid genome rather than the nuclear genome. A recombinant antibody that comprises the heavy- and light-chain variable regions joined by a flexible peptide linker. The linker is long enough to allow separation of the domains so that two of the polypeptides can assemble into a dimer, making the antibody divalent. A recombinant antibody in which the heavy- and light-chain variable regions are part of the same polypeptide chain, which also includes the heavy-chain hinge region and one heavy-chain constant domain.

Usually leaves of tobacco although many other species can be used that are transiently transformed with Agrobacterium tumefaciens , which results in the transient expression of recombinant proteins. This is a useful strategy for testing expression constructs and obtaining small amounts of protein for analysis before going to the expense of transgenics. Broad-leaf flowering plants the seeds of which contain two cotyledons embryonic seed leaves that either remain in the seed when the plant germinates or emerge and become green. Examples include potato, tomato, tobacco and all peas and beans.

Narrow-leaf plants the seeds of which contain one cotyledon. Examples include cereals, grasses, orchids and lilies. A single antigenic determinant on a protein that is recognized by an antibody.


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A single protein can have many epitopes. Interstitial cells in the testis that are responsible for the production of male sex hormones, such as testosterone, and are important in male sexual differentiation. An in vitro mutagenesis procedure that is often carried out using the polymerase chain reaction in which specific mutations are introduced into a DNA molecule. A short sequence of mainly hydrophobic amino acids at the N-terminus of secreted proteins. This peptide is captured by a signal-recognition particle as it emerges from the ribosome, which allows the ribosome to be transported to the endoplasmic reticulum.

There are several recombinant proteins derived from plants that were the basic idea of edible vaccines, directly eaten as fruits tomatoes and bananas and vegetables lettuce and carrots ; accordingly, no processing costs will be demanded by the elimination of processing, [ 66 ]. Bananas, as a fruit host in agricultural products, have particularly attracted lots of customers for the production of edible vaccines, especially for developing countries. This has been widely developed in such countries because of long distance transports and cooling requirements [ 42 ].

Apart from the mentioned advantages, high digestibility and palatability of bananas have won a wide public acceptance for the vaccination of children [ 67 , 68 ]. The sufficiency of potatoes, eaten in raw or low processed forms, for edible vaccines has resulted in their wide production. Potatoes, like seeds, have the advantage of production stability due to a special molecular environment allocated in glands [ 69 ].

The risks of transgenic plants are divided into two categories: one category directly affects humans and the other endangers environment and other organisms. The attack of immune system can disable these medicines and lead to the stimuli for the allergic reactions, some of which have been elaborated as follows:. There are some concerns in terms of environmental pollution about the entrance of transgenes into the food chain, which requires a sound management and supervision.

The other concern refers to the grain transformations using agrobacterium since grains are important crops in the production of pharmaceutical protein. The reactions of immune system can disable the medicines produced in plants and be the stimuli for allergic reactions [ 70 ]. The development stages and subsequent commercialization of the products is the subject of consideration in the second phase of clinical trials. A number of small biotechnology companies have aimed to commercialize the antibodies produced in plants.

While there have been great advances in the field of biomedicine production in plants on large scales, fundamental studies are demanded to pave the way for the commercialization of these products. The aim of molecular farming is to produce large quantities of active and secure pharmaceutical proteins with lower prices. With the scientific advances in the field of bio-technology, nowadays, gene transfer methods in plants have considerably developed.

These transgenic plants in comparison with other microbial and animal expression systems have various advantages in terms of easy production, cost, safety, etc. So far, lots of valuable pharmaceutical proteins and antibodies have been produced by the help of this method, which remarkably has helped the treatment of patients especially in developing countries where the production and preservation costs of such medicines cannot be afforded.

Manufacturing of Pharmaceutical Proteins: From Technology to Economy

However, there are some disputes, such as public acceptance, transgene escape and biosecurity, clinical and commercialization investigations of products, etc. We are grateful to the Azarbaijan Shahid Madani University, especially the Vice president for research since financial assistance for some research in this field. Licensee IntechOpen. This chapter is distributed under the terms of the Creative Commons Attribution 3. Help us write another book on this subject and reach those readers. Login to your personal dashboard for more detailed statistics on your publications. Edited by Hany El-Shemy.

We are IntechOpen, the world's leading publisher of Open Access books. Built by scientists, for scientists. Our readership spans scientists, professors, researchers, librarians, and students, as well as business professionals. Downloaded: Abstract Plant molecular farming describes the production of recombinant proteins and other secondary metabolites in plants. Keywords Molecular farming transgenic plants biomedicines protein stability. Introduction Molecular farming is a biotechnological program that includes the genetic modification of agricultural products to produce proteins and chemicals for commercial and pharmaceutical purposes.

The strategies of plant transformation Plant molecular farming depending on the production of transgenic plants has been operated by two general methods as the following: 2. Stable or permanent expression systems a. Temporary or transient expression systems A transient production may be the fastest system for plant molecular farming [ 21 ].

The given systems include the following methods: Agrobacterium transformation method: Infiltration of recombinant agrobacterium suspension into tobacco leaf tissue is achieved without stable gene transfer, which facilitates the transfer of T-DNA to a very high percentage of cells, where the transgenes are expressed at a high level without a stable transfer of genes.

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The advantages of utilizing transgenic plants as bioreactors Comparison of different expression systems see Table 1 reveals the advantages of plants in comparison with other expression systems as follows: The healthiness of derived products plants cannot be the host of human pathogens and bacterial toxins. Table 1. The limitations and optimization of plant production systems 4.

The problem of product shortage or the same recombinant proteins 4. Optimization of expression of transcripts To optimize the expression of transcripts, a widely used strategy is the use of building promoters, such as cauliflower mosaic virus 35S RNA promoter and maize 1-ubiquitin promoter, respectively, suitable for spilt-cotyledons and single-cotyledons [ 27 ].

Optimization of translation Expression constructs can be designed for guaranteeing the efficiency of translation and the sustainability of transcripts. Optimization of protein stability To optimize the stability of recombinant proteins, known as the most important limiting factor for the function of molecular farming [ 34 ], the targeting of proteins into certain intracellular parts is demanded.

Targeting of proteins can be done by the following pathways and organelles, The intracellular parts, like protein storage vacuoles, have been discovered for the accumulation of recombinant proteins [ 30 ].

Behme, Stefan

Challenge of glycosylation protein quality Glycosylation refers to the covalent binding of sugars to proteins in order to increase close-packing, biological activity, solubility, and biological functionality [ 5 ]. Prohibiting the activity of fucosyltransferase and xylyltransferase enzymes. Selecting appropriate host plants Major economical factors in appointing an appropriate host include the total biomass yield, storage characteristics, ease of transport, value of recombinant proteins, maintenance costs, its availability for workers, required area, duration of production cycle, cost of subsequent products, and edibility [ 27 , 34 ].

Predicting the intracellular localization of the recombinant protein The importance of intracellular localization of proteins is due to the functional consequences of proteins.

Proteins and biomedicines produced in plants Plants are able to produce those bacterial and viral recombinant antigens that preserve the capability of making the structures Type IV similar to those witnessed in mammalian systems, and the post-translational modifications are operated to maintain the biological activity of proteins. The biomedicines produced in plants are as follows: Antigens for the production of edible vaccines: Antigens, used for generating an immune response resulting in immunity against diseases in human proteins, are expressed from different pathogens in plants.

Molecular farming and metabolic engineering, an opportunity for producing plants with a high technology Molecular farming and metabolic engineering make the production of new high-tech products possible. Purification and downstream processing of the recombinant proteins Recovery usually includes the process and breakdown of plant tissues, protein extraction, solid-liquid separation, and protein concentration while purification encompasses safety protection, liquid-liquid extraction, membrane filtration, chromatography, etc.

Bio-safety and the challenges in the domain of protein production and biomedicines in molecular farming The risks of transgenic plants are divided into two categories: one category directly affects humans and the other endangers environment and other organisms. The attack of immune system can disable these medicines and lead to the stimuli for the allergic reactions, some of which have been elaborated as follows: There are some concerns in terms of environmental pollution about the entrance of transgenes into the food chain, which requires a sound management and supervision.

Perspectives upon the commercial production of medicines and pharmaceutical proteins in molecular farming The development stages and subsequent commercialization of the products is the subject of consideration in the second phase of clinical trials. Conclusion The aim of molecular farming is to produce large quantities of active and secure pharmaceutical proteins with lower prices.

Acknowledgments We are grateful to the Azarbaijan Shahid Madani University, especially the Vice president for research since financial assistance for some research in this field. More Print chapter. How to cite and reference Link to this chapter Copy to clipboard.

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