Departmental Papers (Dental)
Document Type
Journal Article
Date of this Version
2009
Publication Source
Current Topics in Microbiology and Immunology
Volume
332
Start Page
33
Last Page
54
DOI
10.1007/978-3-540-70868-1_3
Abstract
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. Oral delivery is facilitated by hyperexpression 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. Hyperexpression of several therapeutic proteins, including human serum albumin (11.1% TSP), somatotropin (7% TSP), interferon-alpha (19% TSP), interferon-gamma (6% TSP), and antimicrobial peptide (21.5% TSP), facilitates efficient and economic purification. Also, the presence of chaperones and enzymes in chloroplasts facilitates assembly of complex multisubunit 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. Purification of human proinsulin has been achieved using novel purification strategies (inverse temperature transition property) that do not require expensive column chromatography techniques. Thus, transgenic chloroplasts are ideal bioreactors for production of functional human and animal therapeutic proteins in an environmentally friendly manner.
Copyright/Permission Statement
The final publication is available at Springer via http://dx.doi.org/10.1007/978-3-540-70868-1_3
Recommended Citation
Chebolu, S., & Daniell, H. (2009). Chloroplast-Derived Vaccine Antigens and Biopharmaceuticals: Expression, Folding, Assembly and Functionality. Current Topics in Microbiology and Immunology, 332 33-54. http://dx.doi.org/10.1007/978-3-540-70868-1_3
Date Posted: 01 March 2022
Comments
At the time of publication, author Henry Daniell was affiliated with the Department of Molecular Biology and Microbiology at the University of Central Florida. Currently, he is a faculty member at the School of Dental Medicine at the University of Pennsylvania