Expression and Affinity Purification of Recombinant Proteins from Plants

Desai, Urvee A.; Sur, Gargi; Daunert, Sylvia; Babbitt, R. A.; Li, Qingshun Quinn

doi:10.1006/prep.2002.1627

Cited by 21 publications

(14 citation statements)

References 20 publications

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“…However, the presence of vascular tissue in mature leaves can make grinding more difficult. Another widely used method at the small scale is the grinding of frozen leaves in liquid nitrogen to a dry powder and resuspension in an aqueous buffer for protein release (Blaszczyk et al ., 2002; Desai et al ., 2002). Grinding in liquid nitrogen is unlikely to be a feasible option at large scale because of high cost implications.…”

Section: Discussionmentioning

confidence: 99%

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Hassan

Dolleweerd

Ioakeimidis

et al. 2008

Plant Biotechnology Journal

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SummaryMonoclonal antibody production from transgenic tobacco plants offers many advantages over other heterologous production systems, creating the prospect of production at a scale that will allow new prophylactic and therapeutic applications in global human and animal health. However, information on the major processing factors to consider for large-scale purification of antibodies from transgenic plants is currently limited, and is in urgent need of attention. The purpose of this project was to investigate methods for the initial extraction of recombinant immunoglobulin G (IgG) antibodies from transgenic tobacco leaf tissue.Three different transgenic plant lines were studied in order to establish the parameters for optimal extraction of monoclonal antibodies that accumulate in the apoplasm, at the plasma membrane or within the endoplasmic reticulum. For each transgenic line, seven techniques for physical extraction were compared. The factors that determine the optimal extraction of antibodies from plants have a direct influence on the initial choice of expression strategy, and so must be considered at an early stage. The use of small-scale techniques that are applicable to large-scale purification was a particularly important consideration. The optimal extraction technique varied with the target location of IgG in the plant cell, and the dependence of antibody yield on the physical extraction methodology employed, the pH of the extraction buffer and the extraction temperature was demonstrated in each case. The addition of detergent to the extraction buffer may improve the yield, but this was found to be dependent on the site of accumulation of IgG within the plant cell.

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Section: Discussionmentioning

confidence: 99%

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Hassan

Dolleweerd

Ioakeimidis

et al. 2008

Plant Biotechnology Journal

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“…Из регуляторных белков, помимо уже упомянутого соматотропина, в растениях табака синтезировали эн-кефалины , эпидермальный фактор роста , кальмодулин (Desai et al, 2002) и инсулиноподобный фактор роста (Daniell et al, 2009), причём последний -в транспластомных растениях. Получены трансгенные растения резухо-видки, в семенах которой накапливается человеческий рекомбинантный инсулин .…”

Section: регуляторные белкиunclassified

Transgenic plants as bioreactors for the production of substances of medicinal and veterinary importance

Savel’eva¹,

Burlakovskiy

Yemelyanov

et al. 2015

Ecological genetics

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The use of plants as bioreactors has become of a great importance in the modern biotechnology. The transgenic plants are capable of synthesizing of many substances, including valuable pharmaceuticals. Plants possess a number of advantages compared to conventional bioreactors - microorganisms and animal cell cultures. The product safety and lower production costs are among them. One of the promising directions in plant biotechnology is the creation of “edible vaccines, plantibodies and adjuvants” based on recombinant antigens, immunoglobulins and immunoregulatory cytokines. Edible bioreactor plants can be administered as food and feed additives in medicine and veterinary avoiding expensive purification procedures. Interferons have antiviral, antibacterial, antitumor and immunomodulatory activity, and are implicated in the prophylaxis and therapy of diseases of different etiologies. Investigations concerning with obtaining of bioreactor plants synthesizing γ-interferons of mammals and birds are carried out in the laboratory of genetic and cellular engineering of plants St. Petersburg State University. Our recent achievements in the creation of inbreed tobacco line producing bovine γ-interferon are described.

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“…By attaching the protein calmodulin (CaM) as an affinity tag, GUS was purified on a phenothiazine affinity column, and 20-fold purification could be achieved (Desai et al 2002). Histag is also a widely used method to enhance protein purification from tobacco extract by IMAC.…”

Section: Product Recovery Examplesmentioning

confidence: 99%

“…For tobacco, the presence of strong veins makes direct homogenization of mature fresh leaves more difficult. One widely used method for small-scale processing is to freeze the leaves and grind them in liquid nitrogen before extraction (Blaszczyk et al, 2002;Desai et al, 2002), but this method has obvious limitations when large amounts of tobacco leaves are to be processed.…”

Section: Product Recovery Examplesmentioning

confidence: 99%

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Menkhaus¹,

Bai²,

Zhang³

et al. 2004

Biotechnol. Prog.

153

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The past 5 years have seen the commercialization of two recombinant protein products from transgenic plants, and many recombinant therapeutic proteins produced in plants are currently undergoing development. The emergence of plants as an alternative production host has brought new challenges and opportunities to downstream processing efforts. Plant hosts contain a unique set of matrix contaminants (proteins, oils, phenolic compounds, etc.) that must be removed during purification of the target protein. Furthermore, plant solids, which require early removal after extraction, are generally in higher concentration, wider in size range, and denser than traditional bacterial and mammalian cell culture debris. At the same time, there remains the desire to incorporate highly selective and integrative separation technologies (those capable of performing multiple tasks) during the purification process from plant material. The general plant processing and purification scheme consists of isolation of the plant tissue containing the recombinant protein, fractionation of the tissue along with particle size reduction, extraction of the target protein into an aqueous medium, clarification of the crude extract, and finally purification of the product. Each of these areas will be discussed here, focusing on what has been learned and where potential concerns remain. We also present details of how the choice of plant host, along with location within the plant for targeting the recombinant protein, can play an important role in the ultimate ease of recovery and the emergence of regulations governing plant hosts. Major emphasis is placed on three crops, canola, corn, and soy, with brief discussions of tobacco and rice.

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Expression and Affinity Purification of Recombinant Proteins from Plants

Cited by 21 publications

References 20 publications

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Considerations for extraction of monoclonal antibodies targeted to different subcellular compartments in transgenic tobacco plants

Transgenic plants as bioreactors for the production of substances of medicinal and veterinary importance

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