The baculovirus expression vector system (BEVS) is a widely used platform for the production of recombinant eukaryotic proteins. However, the BEVS has limitations in comparison to other higher eukaryotic expression systems. First, the insect cell lines used in the BEVS cannot produce glycoproteins with complex‐type N‐glycosylation patterns. Second, protein production is limited as cells die and lyse in response to baculovirus infection. To delay cell death and lysis, we transformed several insect cell lines with an expression plasmid harboring a vankyrin gene (P‐vank‐1), which encodes an anti‐apoptotic protein. Specifically, we transformed Sf9 cells, Trichoplusia ni High FiveTM cells, and SfSWT‐4 cells, which can produce glycoproteins with complex‐type N‐glycosylation patterns. The latter was included with the aim to increase production of glycoproteins with complex N‐glycans, thereby overcoming the two aforementioned limitations of the BEVS. To further increase vankyrin expression levels and further delay cell death, we also modified baculovirus vectors with the P‐vank‐1 gene. We found that cell lysis was delayed and recombinant glycoprotein yield increased when SfSWT‐4 cells were infected with a vankyrin‐encoding baculovirus. A synergistic effect in elevated levels of recombinant protein production was observed when vankyrin‐expressing cells were combined with a vankyrin‐encoding baculovirus. These effects were observed with various model proteins including medically relevant therapeutic proteins. In summary, we found that cell lysis could be delayed and recombinant protein yields could be increased by using cell lines constitutively expressing vankyrin or vankyrin‐encoding baculovirus vectors.
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Research reported in this publication was supported by the National Institute of General Medical Sciences of the National Institute of Health under Award Numbers R42GM075628 and R44GM093411 to ParaTechs. This technology was supported in part by an award from the Kentucky Cabinet for Economic Development, Office of Entrepreneurship, under the Grant Agreement KSTC‐184–512‐07–023 with the Kentucky Science and Technology Corporation to ParaTechs.
Steele, Kendra H.; Stone, Barbara J.; Franklin, Kathleen M.; Fath-Goodin, Angelika; Zhang, Xiufeng; Jiang, Haobo; Webb, Bruce A.; and Geisler, Christoph, "Improving the Baculovirus Expression Vector System with Vankyrin-Enhanced Technology" (2017). Entomology Faculty Publications. 184.