Production of biopharmaceuticals and vaccines in plants via the chloroplast genome.
Transgenic plants offer many advantages, including low cost of production (by elimination of fermenters), storage and transportation; heat stability; and absence of human pathogens. When therapeutic proteins are orally delivered, plant cells protect antigens in the stomach through bioencapsulation and eliminate the need for expensive purification and sterile injections, in addition to development of both systemic and mucosal immunity. Chloroplast genetic engineering offers several advantages, including high levels of transgene expression, transgene containment via maternal inheritance and multi-gene expression in a single transformation event. Hyper-expression of vaccine antigens against cholera, tetanus, anthrax, plague or canine parvovirus (4-31% of total soluble protein, tsp) in transgenic chloroplasts (leaves) or non-green plastids (carrots, tomato), as well as the availability of antibiotic-free selectable markers or the ability to excise selectable marker genes, facilitate oral delivery. Hyper-expression of several therapeutic proteins, including human serum albumin (11.1% tsp), somatotropin (7% tsp), interferon-gamma (6% tsp), anti-microbial peptide (21.5% tsp), facilitates efficient and economic purification. Also, the presence of chaperones and enzymes in chloroplasts facilitate assembly of complex multi-subunit proteins and correct folding of human blood proteins with proper disulfide bonds. Functionality of chloroplast-derived vaccine antigens and therapeutic proteins has been demonstrated by several assays, including the macrophage lysis assay, GM1-ganglioside binding assay, protection of HeLa cells or human lung carcinoma cells against encephalomyocarditis virus, systemic immune response, protection against pathogen challenge, and growth or inhibition of cell cultures. Thus, transgenic chloroplasts are ideal bioreactors for production of functional human and animal therapeutic proteins in an environmentally friendly manner.
Daniell
Henry
H
Department of Molecular Biology and Microbiology, University of Central Florida, Biomolecular Science, Orlando, FL 32816-2364, USA. daniell@mail.ucf.edu
D001702:Biopharmaceutics / Q000379:methods; D002736:Chloroplasts / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D018745:Genome, Plant / Q000235:genetics*; D004364:Pharmaceutical Preparations / Q000378:metabolism*; D010936:Plant Extracts / Q000235:genetics / Q000627:therapeutic use; D030821:Plants, Genetically Modified / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism*; D014612:Vaccines / Q000235:genetics / Q000378:metabolism*
D004364:Pharmaceutical Preparations; D010936:Plant Extracts; D011994:Recombinant Proteins; D014612:Vaccines