FFATA: Collaborative Research: Development of an Integrated Algae Platform for Affordable Protein Therapeutic and Vaccine Molecules
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Scalable and low-cost production platforms are of growing importance for the development of affordable recombinant protein molecules. This is especially true in the case of complex recombinant proteins that are often difficult and expensive to produce in commercial protein production platforms, such as mammalian cell culture and E. coli. Protein production by mammalian cell cultures is still expensive because of stringent nutritional and growth requirements, specialized bioreactor design and environmental conditions, coupled with often low expression levels. E. coli, although able to produce large quantities of protein inexpensively, is generally inefficient at producing properly folded, complex proteins as soluble molecules. The single-cell eukaryotic alga, Chlamydomonas reinhardtii, has simple growth requirements (light, carbon dioxide, and salts) and its use as a production platform for complex recombinant proteins combines many of the most advantageous features of fermentation and cell cultures. The specific features of C. reinhardtii chloroplast expression include: rapid generation of stably transformed master cell lines, efficient mammalian protein folding in a non-reducing environment, a host metabolism that is tolerant of "difficult-to-express" proteins, a low propensity for undesirable post-translational modifications and formation of insoluble aggregates, cell doubling times under 10 h using completely defined protein-free inorganic media, demonstrated scalability, and absence of endotoxin and mammalian virus contamination. The investigators have already demonstrated that C. reinhardtii is able to produce complex, multi-domain proteins that cannot be efficiently expressed in and purified from other heterologous systems. This proposal focuses on the expression and purification of two unique and therapeutically important recombinant proteins that will serve to demonstrate the utility of the platform, and that could have high-impact provided they can be produced and purified at a competitive cost. To develop C. reinhardtii as a broad and competitive platform for producing a unique class of recombinant proteins, the interdisciplinary team with complementary expertise will address challenges related to protein production, recovery, and purification. The goal of the proposed research is to develop technologies and processes for an efficient, rapid, and scalable recombinant protein production platform for using C. reinhardtii.