Gu Zhengrong
Agricultural and Biosystems Engineering, South Dakota State University, North Campus Drive 1400, Brookings, SD, 57007, USA,
Methods Mol Biol. 2014;1129:77-87. doi: 10.1007/978-1-62703-977-2_8.
Transgenic plants producing recombinant proteins are a potential low-cost substitute for complex bioreactors. However, the development of new robust downstream processes to improve protein recovery and isolation from plant feedstock is critical to promote this new technique because downstream processing costs typically contribute to more than 80 % of the total cost. Polishing separation platforms for protein purification, such as chromatography and membrane filtration, have been well established, while little attention has been allocated to initial concentration and separation procedures. In this chapter, application of aqueous two-phase partitioning (ATPP), as an attractive alternative to traditional processes for recovering and isolating target proteins from plant green tissues or seeds, as well as the main advances reported in literature concerning ATPP for the isolation and purification of proteins from plant feedstock are reviewed. The potential application of ATPP as an integrated extraction and isolation step and isolation step after extraction or protein characterization method is discussed separately. The connection of ATPP with traditional protein separation processes is discussed. The separation mechanisms of ATPP are explained based on surface properties of proteins and polymer systems. Finally, the future trends in applying ATPP for protein separation are discussed.
生产重组蛋白的转基因植物是复杂生物反应器潜在的低成本替代品。然而，开发新的稳健下游工艺以提高从植物原料中回收和分离蛋白质的能力对于推广这项新技术至关重要，因为下游加工成本通常占总成本的80%以上。用于蛋白质纯化的精制分离平台，如色谱法和膜过滤，已经得到了很好的确立，而对初始浓缩和分离程序的关注却很少。在本章中，综述了双水相分配（ATPP）作为从植物绿色组织或种子中回收和分离目标蛋白的传统方法的一种有吸引力的替代方法的应用，以及文献中报道的关于ATPP从植物原料中分离和纯化蛋白质的主要进展。分别讨论了ATPP作为集成提取和分离步骤以及提取后或蛋白质表征方法后的分离步骤的潜在应用。讨论了ATPP与传统蛋白质分离工艺的联系。基于蛋白质和聚合物系统的表面性质解释了ATPP的分离机制。最后，讨论了将ATPP应用于蛋白质分离的未来趋势。
