Identification of two subtilisin-like serine proteases engaged in the degradation of recombinant proteins in Nicotiana benthamiana.
Alejandro A Puchol TarazonaDaniel MareschAnnette GrillJanet BakalarzJuan A Torres AcostaAlexandra CastilhoHerta SteinkellnerLukas MachPublished in: FEBS letters (2020)
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 α1 -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.
Keyphrases
- big data
- antiretroviral therapy
- poor prognosis
- hiv infected
- human immunodeficiency virus
- hiv positive
- protein kinase
- binding protein
- cell proliferation
- hepatitis c virus
- electronic health record
- room temperature
- small molecule
- cell free
- single cell
- hiv testing
- high throughput
- metal organic framework
- protein protein
- transition metal
- transcription factor