Identification of two subtilisin‐like serine proteases engaged in the degradation of recombinant proteins in Nicotiana benthamiana. Issue 3 (11th December 2020)

• Record Type: Journal Article
• Authors: Puchol Tarazona, Alejandro A.; Maresch, Daniel; Grill, Annette; Bakalarz, Janet; Torres Acosta, Juan A.; Castilho, Alexandra; Steinkellner, Herta; Mach, Lukas
• Is Part Of: FEBS letters. Volume 595:Issue 3(2021)
• Page Start: 379
• Page End: 388
• Publication Date: 2020-12-11
• Subjects: biopharmaceutical -- molecular farming -- monoclonal antibody -- Nicotiana benthamiana -- recombinant protein expression -- serine protease; Biochemistry -- Periodicals; Biophysics -- Periodicals; Molecular biology -- Periodicals; Biochimie -- Périodiques; Biochemistry; Biophysics; Molecular biology; Periodicals; 572.05
• DOI: 10.1002/1873-3468.14014
• Languages: English
• ISSNs: 0014-5793

Abstract

Abstract : 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. Abstract :