Recombinant Protein Expression in Nicotiana
Recombinant Protein Expression in Nicotiana
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DOI：
10.1007/978-1-61737-957-4_11
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发表时间：
2011-01-01
期刊：
影响因子：
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通讯作者：
Palmer, Kenneth E.
中科院分区：
文献类型：
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作者：
Matoba, Nobuyuki;Davis, Keith R.;Palmer, Kenneth E.
重组蛋白药物现在广泛用于慢性疾病的治疗，一些重组蛋白亚单位疫苗被批准用于人类和兽医。随着对单克隆抗体等复杂蛋白质药物的需求不断增长，生产能力变得有限。对基于哺乳动物细胞培养的制造系统的安全、可扩展和经济替代品的需求日益增加，这需要大量的资本投资来建造新的制造设施。自从1989年发表了一篇报道转基因植物中免疫球蛋白表达的开创性论文以来，植物表达系统已经取得了许多技术进步，到目前为止，在非粮食作物（如烟草）的叶组织中生产蛋白质被认为是一种可行的替代方法。特别是，源自重组植物病毒载体的瞬时表达系统为快速表达筛选、构建优化和表达规模扩大提供了机会。从烟叶组织中提取重组蛋白可以通过收集分泌蛋白部分来实现，也可以通过用缓冲液研磨叶片后的总蛋白提取物来实现。从固体分离后，主要的纯化挑战是光合复合体元素的污染，这可以通过应用各种简单和成熟的策略来解决。总之，在烟叶组织中安全、高效、可扩展地生产重组蛋白所需的技术已经成熟，一些产品已经进入I期临床试验，很快就会有丰富的重组疫苗、杀微生物剂和治疗蛋白的管道。
Recombinant protein pharmaceuticals are now widely used in treatment of chronic diseases, and several recombinant protein subunit vaccines are approved for human and veterinary use. With growing demand for complex protein pharmaceuticals, such as monoclonal antibodies, manufacturing capacity is becoming limited. There is increasing need for safe, scalable, and economical alternatives to mammalian cell culture-based manufacturing systems, which require substantial capital investment for new manufacturing facilities. Since a seminal paper reporting immunoglobulin expression in transgenic plants was published in 1989, there have been many technological advances in plant expression systems to the present time where production of proteins in leaf tissues of nonfood crops such as Nicotiana species is considered a viable alternative. In particular, transient expression systems derived from recombinant plant viral vectors offer opportunities for rapid expression screening, construct optimization, and expression scale-up. Extraction of recombinant proteins from Nicotiana leaf tissues can be achieved by Collection of secreted protein fractions, or from a total protein extract after grinding the leaves with buffer. After separation from solids, the major purification challenge is contamination with elements of the photosynthetic complex, which can be solved by application of a variety of facile and proven strategies. In conclusion, the technologies required for safe, efficient, scalable manufacture of recombinant proteins in Nicotiana leaf tissues have matured to the point where several products have already been in phase I clinical trials and will soon be followed by a rich pipeline of recombinant vaccines,microbicides, and therapeutic proteins.