Ocampo Carolina Gabriela, Petruccelli Silvana
CIDCA-CCT-La Plata CONICET, Facultad de Ciencias Exactas-Universidad Nacional de La Plata (UNLP), La Plata, Argentina.
Methods Mol Biol. 2018;1789:65-80. doi: 10.1007/978-1-4939-7856-4_6.
Plant-based platforms are extensively use for the expression of recombinant proteins, including monoclonal antibodies (mAbs). Generally, immunoglobulins (Igs) are sorted to the apoplast, which is often afflicted with intense proteolysis. Here, we describe methods to transiently express mAbs sorted to central vacuole in Nicotiana benthamiana leaves and to characterize the obtained IgG. Central vacuole is an appropriate compartment for the efficient production of Abs, consequently vacuolar sorting should be considered as an alternative strategy to obtain high protein yields.
基于植物的平台被广泛用于重组蛋白的表达，包括单克隆抗体（mAb）。一般来说，免疫球蛋白（Ig）会被分选到质外体，而质外体常常遭受强烈的蛋白水解作用。在这里，我们描述了在本氏烟草叶片中瞬时表达分选到中央液泡的单克隆抗体的方法，并对获得的IgG进行表征。中央液泡是高效生产抗体的合适区室，因此液泡分选应被视为获得高蛋白产量的一种替代策略。
