The microalga<i>Chlamydomonas reinhardtii</i>as a platform for the production of human protein therapeutics

Rasala, Beth A.; Mayfield, Stephen P.

doi:10.4161/bbug.2.1.13423

Cited by 106 publications

(64 citation statements)

References 33 publications

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“…In addition to energy production, microalgae can produce a variety of other bioproducts including nutraceuticals and recombinant proteins such as industrial enzymes or therapeutics (3,4). Using the green algae Chlamydomonas reinhardtii, we have demonstrated that algae are capable of expressing, folding, and accumulating a range of human therapeutic proteins in the chloroplast (5-7).…”

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confidence: 99%

Production of unique immunotoxin cancer therapeutics in algal chloroplasts

Tran

Van

Barrera

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

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The idea of targeted therapy, whereby drug or protein molecules are delivered to specific cells, is a compelling approach to treating disease. Immunotoxins are one such targeted therapeutic, consisting of an antibody domain for binding target cells and molecules of a toxin that inhibits the proliferation of the targeted cell. One major hurdle preventing these therapies from reaching the market has been the lack of a suitable production platform that allows the cost-effective production of these highly complex molecules. The chloroplast of the green alga Chlamydomonas reinhardtii has been shown to contain the machinery necessary to fold and assemble complex eukaryotic proteins. However, the translational apparatus of chloroplasts resembles that of a prokaryote, allowing them to accumulate eukaryotic toxins that otherwise would kill a eukaryotic host. Here we show expression and accumulation of monomeric and dimeric immunotoxin proteins in algal chloroplasts. These fusion proteins contain an antibody domain targeting CD22, a B-cell surface epitope, and the enzymatic domain of exotoxin A from Pseudomonas aeruginosa. We demonstrated that algal-produced immunotoxins accumulate as soluble and enzymatically active proteins that bind target B cells and efficiently kill them in vitro. We also show that treatment with either the mono-or dimeric immunotoxins significantly prolongs the survival of mice with implanted human B-cell tumors.

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confidence: 99%

Production of unique immunotoxin cancer therapeutics in algal chloroplasts

Tran

Van

Barrera

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

Self Cite

165

118

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“…These factors raise the possibility that cultures can be grown in large-scale photobioreactors, which significantly reduces the risk of contamination and the escape of genetic modified strains to the environment, and makes rapid scale-up possible. In addition, green algae fall into the GRAS (generally regarded as safe) category, potentially eliminating some downstream processing steps associated with transgenically produced therapeutics (Rasala and Mayfield, 2011;Specht et al, 2010).…”

Section: Introductionmentioning

confidence: 99%

Expression and membrane-targeting of an active plant cytochrome P450 in the chloroplast of the green alga Chlamydomonas reinhardtii

et al. 2015

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The unicellular green alga Chlamydomonas reinhardtii has potential as a cell factory for the production of recombinant proteins and other novel compounds, but mainstream adoption has been hindered by a scarcity of genetic tools and a need to identify products that can be generated in a cost-effective manner. A promising strategy is to use algal chloroplasts as a site for synthesis of high value bioactive compounds such as diterpenoids since these are derived from metabolic building blocks that occur naturally within the organelle. However, synthesis of these complex plant metabolites requires the introduction of membraneassociated enzymes including cytochrome P450 enzymes (P450s). Here, we show that a gene (CYP79A1) encoding a model P450 can be introduced into the C. reinhardtii chloroplast genome using a simple transformation system. The gene is stably expressed and the P450 is efficiently targeted into chloroplast membranes, where it is active and accounts for 0.4% of total cell protein. These results provide proof of concept for the introduction of diterpenoid synthesis pathways into the chloroplast of Chlamydomonas reinhardtii.

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“…Chlamydomonas reinhardtii is the most successfully used microalgae, because it is genetically well characterized with all three genomes (the nuclear, chloroplast and mitochondrial) sequenced, and genetic transformation methods are well established (Rasala & Mayfield, 2011). Proteins such as human antibodies (Mayfield & Franlikn, 2005), human glutamic acid decarboxylase 65 (hGAD65) (Wang et al, 2008), domain 14 of fibronectin (14FN3), VEGF and HMGB1 (Rasala & Mayfield, 2011) have been produced in C. reinhardtii.…”

Section: Emerging Plant-based Systemsmentioning

confidence: 99%