Title
Chloroplast-Derived Vaccine Antigens And Biopharmaceuticals: Expression, Folding, Assembly And Functionality
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 chap-erones 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 bio-reactors for production of functional human and animal therapeutic proteins in an environmentally friendly manner. © 2009 Springer-Verlag Berlin Heidelberg.
Publication Date
9-24-2010
Publication Title
Current Topics in Microbiology and Immunology
Volume
332
Number of Pages
33-54
Document Type
Review
Personal Identifier
scopus
DOI Link
https://doi.org/10.1007/978-3-540-70868-1_3
Copyright Status
Unknown
Socpus ID
67849122688 (Scopus)
Source API URL
https://api.elsevier.com/content/abstract/scopus_id/67849122688
STARS Citation
Chebolu, S. and Daniell, H., "Chloroplast-Derived Vaccine Antigens And Biopharmaceuticals: Expression, Folding, Assembly And Functionality" (2010). Scopus Export 2010-2014. 1179.
https://stars.library.ucf.edu/scopus2010/1179