Chloroplast-derived vaccine antigens and biopharmaceuticals: expression, folding, assembly and functionality.
Chloroplast-derived vaccine antigens and biopharmaceuticals: expression, folding, assembly and functionality.
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DOI：
10.1007/978-3-540-70868-1_3
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发表时间：
2009
影响因子：
--
通讯作者：
Daniell, H.
中科院分区：
文献类型：
--
作者：
Chebolu, S.;Daniell, H.
叶绿体基因工程提供了几个优点，包括高水平的转基因表达，转基因遏制通过母系遗传，和多基因表达在一个单一的转化事件。通过在转基因叶绿体（叶）或非绿色质体（胡萝卜、番茄）中过表达针对霍乱、破伤风、炭疽、鼠疫或犬细小病毒的疫苗抗原（总可溶性蛋白，TSP的4%-31%）以及无抗生素选择标记的可用性或切除选择标记基因的能力来促进口服递送。几种治疗性蛋白质的过表达，包括人血清白蛋白（11.1%TSP）、生长激素（7%TSP）、干扰素-α（19%TSP）、干扰素-γ（6%TSP）和抗菌肽（21.5%TSP），有助于有效和经济的纯化。此外，叶绿体中的Chaperones和酶的存在促进了复杂的多亚基蛋白质的组装和具有适当二硫键的人血蛋白的正确折叠。叶绿体衍生的疫苗抗原和治疗性蛋白质的功能已经通过几种测定法证明，包括巨噬细胞裂解测定法、GM 1-神经节苷脂结合测定法、针对脑心肌炎病毒的HeLA细胞或人肺癌细胞的保护、全身免疫应答、针对病原体攻击的保护以及细胞培养物的生长或抑制。人胰岛素原的纯化已经实现了使用新的纯化策略（逆温度转换特性），不需要昂贵的柱色谱技术。因此，转基因叶绿体是以环境友好的方式生产功能性人类和动物治疗蛋白的理想生物反应器。
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.
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影响因子：
5.6
作者：
Daniell, H;Lee, SB;Panchal, T;Wiebe, PO
通讯作者：
Wiebe, PO
DOI：
10.1006/bbrc.1999.0584
发表时间：
1999-05-19
影响因子：
3.1
作者：
Figueroa-Soto, CG;Lopez-Cervantes, G;Valenzuela-Soto, EM
通讯作者：
Valenzuela-Soto, EM
影响因子：
3.7
作者：
Gómez, N;Carrillo, C;Escribano, JM
通讯作者：
Escribano, JM
影响因子：
56.9
作者：
HAQ, TA;MASON, HS;ARNTZEN, CJ
通讯作者：
ARNTZEN, CJ
影响因子：
46.9
作者：
Giddings, G;Allison, G;Carter, A
通讯作者：
Carter, A