Green factory: Plants as bioproduction platforms for recombinant proteins

Arkansas Biosciences Institute, Arkansas State University, Jonesboro, AR 72401, United States.
Biotechnology advances (Impact Factor: 8.91). 09/2011; 30(5):1171-84. DOI: 10.1016/j.biotechadv.2011.08.020
Source: PubMed

ABSTRACT Molecular farming, long considered a promising strategy to produce valuable recombinant proteins not only for human and veterinary medicine, but also for agriculture and industry, now has some commercially available products. Various plant-based production platforms including whole-plants, aquatic plants, plant cell suspensions, and plant tissues (hairy roots) have been compared in terms of their advantages and limits. Effective recombinant strategies are summarized along with descriptions of scalable culture systems and examples of commercial progress and success.

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Available from: Maureen Dolan, Dec 15, 2014
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    • "An acceptable product shelf-life is also critical for logistics and inventory management, both for the manufacturer and for the end-user. Even so, it is estimated that with 1% protein expression and 50% protein recovery from purification, the cost of plant based protein is 10 and 1000 fold lower than microbial and mammalian based expression systems, respectively (Xu et al., 2012). Post-licensing activities should also be considered. "
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    ABSTRACT: The production of recombinant vaccines in plants may help to reduce the burden of veterinary diseases, which cause major economic losses and in some cases can affect human health. While there is abundant research in this area, a knowledge gap exists between the ability to create and evaluate plant-based products in the laboratory, and the ability to take these products on a path to commercialization. The current report, arising from a workshop sponsored by the Organisation for Economic Co-operation and Development (OECD) Co-operative Research Programme, addresses this gap by providing guidance in planning for the commercialization of plant-made vaccines for animal use. It includes relevant information on developing business plans, assessing market opportunities, manufacturing scale-up, financing, protecting and using intellectual property, and regulatory approval with a focus on Canadian regulations. Copyright © 2015. Published by Elsevier Inc.
    Biotechnology advances 07/2015; DOI:10.1016/j.biotechadv.2015.07.007 · 8.91 Impact Factor
    • "This study suggests that recombinant protein expression is product-specific and needs to be optimized individually. genic plants have gained significant attention in the last few years as a potential host for recombinant proteins due to the high levels of protein achieved and the low cost of cultivation [5] [6]. Nevertheless, critical issues arise from the lack of transgene containment and possible allergic reactions to plant antigens. "
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    ABSTRACT: Microalgae have potential as platforms for the synthesis of high-value recombinant proteins due to their many beneficial attributes including ease of cultivation, lack of pathogenic agents, and low-cost downstream processing. However, current recombinant protein levels are low compared to microbial platforms and stable insertion of transgenes is available in only a few microalgal strains. We have explored different strategies aimed at increasing growth rate and recombinant protein production in the Chlamydomonas reinhardtii chloroplast. A novel fluorescent protein (vivid Verde Fluorescent Protein, VFP) was expressed under the control of the native atpA promoter/5'UTR element. VFP levels were detected by western blotting, with increased protein levels observed when co-expressed with a gene encoding the Escherichia coli Spy chaperone. We used these transformant lines to study the effect of temperature, light and media on recombinant protein production and cell growth. VFP levels and fluorescence, assessed by flow cytometry, allowed a determination of improved cultivation conditions as 30 °C under mixotrophic mode. These conditions were tested for the accumulation of an antimicrobial endolysin (Cpl-1) of potential commercial interest, observing the outcome obtained for VFP could not be easily replicated for Cpl-1. This study indicates that recombinant protein expression is product-specific and needs to be optimised individually. Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
    Biotechnology Journal 06/2015; 10(8). DOI:10.1002/biot.201400566 · 3.71 Impact Factor
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    • "Transient expression in N. benthamiana leaves usually relies on the Agrobacterium tumefaciens-mediated delivery of T-DNA located transgenes to the nucleus of infiltrated leaf cells, with or without subsequent cell-to-cell or systemic spreading (Giritch et al., 2006; Kapila et al., 1997; Marillonnet et al., 2004). Compared to seeds, leaves are less convenient for storage, necessitating extraction immediately after harvesting (Xu et al., 2012). However, because of the transient nature of the platform, recombinant proteins can be harvested as early as 5 days postinfiltration (Tremblay et al., 2010). "
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    ABSTRACT: VHHs or nanobodies are widely acknowledged as interesting diagnostic and therapeutic tools. However, for some applications, multivalent antibody formats, such as the dimeric VHH-Fc format, are desired to increase the functional affinity. The scope of this study was to compare transient expression of diagnostic VHH-Fc antibodies in Nicotiana benthamiana leaves with their stable expression in Arabidopsis thaliana seeds and Pichia pastoris. To this end, VHH-Fc antibodies targeting green fluorescent protein or the A. thaliana seed storage proteins (albumin and globulin) were produced in the three platforms. Differences were mainly observed in the accumulation levels and glycosylation patterns. Interestingly, although in plants oligomannosidic N-glycans were expected for KDEL-tagged VHH-Fcs, several VHH-Fcs with an intact KDEL-tag carried complex-type N-glycans, suggesting a dysfunctional retention in the endoplasmic reticulum. All VHH-Fcs were equally functional across expression platforms and several outperformed their corresponding VHH in terms of sensitivity in ELISA.
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