Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Austria.
Department of Chemistry, University of Natural Resources and Life Sciences, Vienna, Austria.
FEBS Lett. 2021 Feb;595(3):379-388. doi: 10.1002/1873-3468.14014. Epub 2020 Dec 11.
The tobacco variant Nicotiana benthamiana has recently emerged as a versatile host for the manufacturing of protein therapeutics, but the fidelity of many recombinant proteins generated in this system is compromised by inadvertent proteolysis. Previous studies have revealed that the anti-HIV-1 antibodies 2F5 and PG9 as well as the protease inhibitor α -antitrypsin (A1AT) are particularly susceptible to N. benthamiana proteases. Here, we identify two subtilisin-like serine proteases (NbSBT1 and NbSBT2) whose combined action is sufficient to account for all major cleavage events observed upon expression of 2F5, PG9 and A1AT in N. benthamiana. We propose that downregulation of NbSBT1 and NbSBT2 activities could constitute a powerful means to optimize the performance of this promising platform for the production of biopharmaceuticals. DATABASES: NbSBT sequence data are available in the DDBJ/EMBL/GenBank databases under the accession numbers MN534996 to MN535005.
最近，烟草变异种黄花烟（Nicotiana benthamiana）已成为用于制造蛋白类治疗药物的一种通用宿主，但该系统中生成的许多重组蛋白的准确性因意外蛋白水解而受到影响。先前的研究表明，抗 HIV-1 抗体 2F5 和 PG9 以及蛋白酶抑制剂 α-抗胰蛋白酶（A1AT）特别容易受到黄花烟蛋白酶的影响。在这里，我们鉴定了两种枯草杆菌蛋白酶样丝氨酸蛋白酶（NbSBT1 和 NbSBT2），它们的联合作用足以解释在黄花烟中表达 2F5、PG9 和 A1AT 时观察到的所有主要切割事件。我们提出，下调 NbSBT1 和 NbSBT2 的活性可能是优化该有前途的生物制药生产平台性能的一种有效手段。数据库：NbSBT 序列数据可在 DDBJ/EMBL/GenBank 数据库中以 MN534996 至 MN535005 的 accession numbers 获得。
