FlexiBAC: A versatile, open-source baculovirus vector system for protein expression, secretion, and proteolytic processing

Régis P. Lemaitre, Aliona Bogdanova, Barbara Borgonovo, Jeffrey B. Woodruff, David N. Drechsel

Research output: Contribution to journalArticle

2 Scopus citations

Abstract

Background: Baculovirus-mediated expression in insect cells is a powerful approach for protein production. However, many existing methods are time-consuming, offer limited options for protein tagging, and are unsuitable for secreted proteins requiring proteolytic maturation, such as TGF-β family growth factors. Results: To overcome the limitations of traditional baculovirus expression systems, we engineered "FlexiBAC". This system allows recombinant baculovirus formation inside insect cells and reduces the time between initial cloning and protein production to 13 days. FlexiBAC includes 143 shuttle vectors that append combinations of purification tags, fluorescent markers, proteolytic cleavage sites, trafficking signals, and chemical conjugation tags to the termini of the target protein. This system also overexpresses recombinant furin convertase to allow efficient proteolytic processing of secreted proteins. We demonstrate that FlexiBAC can be used to produce high levels of mature, active forms of TGF-β family growth factors, such as Activin A, as well as other proteins that are typically difficult to reconstitute, such as proteins rich in coiled-coil, low complexity, and disordered domains. Conclusions: FlexiBAC is a protein expression system for production of both cytosolic proteins and secreted proteins that require proteolytic maturation. The design of FlexiBAC and its expansive complementary shuttle vector system reduces cloning steps and simplifies baculovirus production.

Original languageEnglish (US)
Article number20
JournalBMC Biotechnology
Volume19
Issue number1
DOIs
StatePublished - Mar 29 2019

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Keywords

  • Baculovirus
  • Intrinsically disordered proteins
  • Protein expression
  • TGF-β

ASJC Scopus subject areas

  • Biotechnology

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