Immune checkpoint inhibitors (ICIs) have achieved huge clinical success. However, many still have limited response rates, and are prohibitively costly. There is a need for effective and affordable ICIs, as well as local manufacturing capacity to improve accessibility, especially to low-to-middle income countries (LMICs). Here, we have successfully expressed three key ICIs (anti-PD-1 Nivolumab, anti-NKG2A Monalizumab, and anti-LAG-3 Relatimab) transiently in Nicotiana benthamiana and Nicotiana tabacum plants. The ICIs were expressed with a combination of different Fc regions and glycosylation profiles. They were characterized in terms of protein accumulation levels, target cell binding, binding to human neonatal Fc receptors (hFcRn), human complement component C1q (hC1q) and various Fcγ receptors, as well as protein recovery during purification at 100 mg- and kg-scale. It was found that all ICIs bound to the expected target cells. Furthermore, the recovery during purification, as well as Fcγ receptor binding, can be altered depending on the Fc region used and the glycosylation profiles. This opens the possibility of using these two parameters to fine-tune the ICIs for desired effector functions. A scenario-based production cost model was also generated based on two production scenarios in hypothetical high- and low-income countries. We have shown that the product accumulation and recovery of plant production platforms were as competitive as mammalian cell-based platforms. This highlights the potential of plants to deliver ICIs that are more affordable and accessible to a widespread market, including LMICs.

中文翻译：

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治愈杀手：烟草植物制造的癌症免疫检查点抑制剂的表达和生产成本计算


免疫检查点抑制剂（ICIs）已取得巨大的临床成功。然而，许多人的反应率仍然有限，而且成本高昂。需要有效且负担得起的 ICI 以及本地制造能力来提高可及性，特别是对于中低收入国家 (LMIC)。在这里，我们成功地在烟草和烟草植物中瞬时表达了三种关键的 ICI（抗 PD-1 Nivolumab、抗 NKG2A Monalizumab 和抗 LAG-3 Relatimab）。 ICI 通过不同 Fc 区和糖基化谱的组合来表达。它们的特征包括蛋白质积累水平、靶细胞结合、与人新生儿 Fc 受体 (hFcRn)、人补体成分 C1q (hC1q) 和各种 Fcγ 受体的结合，以及 100 mg 和 kg 纯化过程中的蛋白质回收率。规模。结果发现，所有 ICI 均与预期靶细胞结合。此外，纯化过程中的回收率以及 Fcγ 受体结合可以根据所使用的 Fc 区域和糖基化谱进行改变。这使得使用这两个参数来微调 ICI 以获得所需的效应器功能成为可能。基于假设的高收入和低收入国家的两种生产情景还生成了基于情景的生产成本模型。我们已经证明，植物生产平台的产品积累和回收与基于哺乳动物细胞的平台一样具有竞争力。这凸显了工厂提供更便宜且更容易进入包括中低收入国家在内的广泛市场的 ICI 的潜力。