Lara Alvaro R, Ramírez Octavio T, Wunderlich Martin
Departamento de Procesos y Tecnología, Universidad Autónoma Metropolitana-Cuajimalpa, Mexico City, Mexico.
Methods Mol Biol. 2012;824:271-303. doi: 10.1007/978-1-61779-433-9_14.
Plasmid DNA (pDNA) is the base for promising DNA vaccines and gene therapies against many infectious, acquired, and genetic diseases, including HIV-AIDS, Ebola, Malaria, and different types of cancer, enteric pathogens, and influenza. Compared to conventional vaccines, DNA vaccines have many advantages such as high stability, not being infectious, focusing the immune response to only those antigens desired for immunization and long-term persistence of the vaccine protection. Especially in developing countries, where conventional effective vaccines are often unavailable or too expensive, there is a need for both new and improved vaccines. Therefore the demand of pDNA is expected to rise significantly in the near future. Since the injection of pDNA usually only leads to a weak immune response, several milligrams of DNA vaccine are necessary for immunization protection. Hence, there is a special interest to raise the product yield in order to reduce manufacturing costs. In this chapter, the different stages of plasmid DNA production are reviewed, from the vector design to downstream operation options. In particular, recent advances on cell engineering for improving plasmid DNA production are discussed.
质粒DNA(pDNA)是开发有望用于对抗多种传染病、后天性疾病和遗传性疾病(包括艾滋病、埃博拉、疟疾以及不同类型的癌症、肠道病原体和流感)的DNA疫苗和基因疗法的基础。与传统疫苗相比,DNA疫苗具有许多优点,如稳定性高、无传染性、能将免疫反应仅集中于免疫所需的那些抗原以及疫苗保护作用持久。特别是在发展中国家,传统的有效疫苗往往无法获得或过于昂贵,因此需要新型和改良型疫苗。所以,预计在不久的将来对pDNA的需求将大幅上升。由于注射pDNA通常只会引发微弱的免疫反应,因此免疫保护需要几毫克的DNA疫苗。因此,提高产品产量以降低制造成本备受关注。在本章中,将回顾质粒DNA生产的不同阶段,从载体设计到下游操作选项。特别讨论了用于提高质粒DNA产量的细胞工程的最新进展。