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HiTrap Protein A HP

HiTrap蛋白A HP

Company: Cytiva
Catalog#: 17-0403-03
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Monoclonal Antibody Purification (Nicotiana benthamiana Plants)
[Abstract]  Plant-based expression systems provide an alternative biomanufacturing platform for recombinant proteins (Matoba et al., 2011). In particular, plant virus-based vectors can overexpress proteins within days in the leaf tissue of Nicotiana benthamiana (N. benthamiana). To overcome the issues of genetic instability and limited infectivity of recombinant viruses, Agrobacterium-mediated delivery of “deconstructed” virus vectors has become the mainstay for the production of large and/or multicomponent proteins, such as immunoglobulin (Ig)G monoclonal antibodies (mAbs). Here, we describe a method of producing human IgG mAbs in N. benthamiana using the tobamoviral replicon vector magnICON®. The vector can express up to a few hundred mg of a ... [摘要]  基于植物的表达系统为重组蛋白提供了替代的生物制造平台(Matoba等人,2011)。特别地,基于植物病毒的载体可以在烟草(Nicotiana benthamiana)(本塞姆氏烟草)的叶组织中天数内过表达蛋白质。为了克服重组病毒的遗传不稳定性和有限的感染性的问题,土壤杆菌介导的"解构的"病毒载体的递送已经成为生产大和/或多组分蛋白如免疫球蛋白的主要方法Ig)G单克隆抗体(mAb)。在这里,我们描述了在N中产生人IgG mAbs的方法。本生烟草使用烟草花叶病毒复制子载体magnICON 。载体在7天内可以表达高达几百mg的mAb/kg叶材料。显示了分别用于广泛中和的抗HIV和抗流感mAbs VRC01和CR6261的代表性病例(Hamorsky等人,2013)。叶组织匀浆,通过过滤和离心澄清提取物。 mAb通过快速蛋白质液相色谱(FPLC)使用蛋白A亲和力和Phenyl HP疏水界面树脂纯化。