Expression of an immunocomplex consisting of Fc fragment fused with a consensus dengue envelope domain III in Saccharomyces cerevisiae

Abstract

Objectives To explore Saccharomyces cerevisiae as an expression platform for dengue oral immune complex vaccine development. Results Molecular engineering was applied to create a fusion gene construct (scEDIII-PIGS) consisting of a yeast codon optimized sequence encoding for a synthetic consensus dengue envelope domain III (scEDIII) followed by a modified IgG Fc domain (PIGS). Northern blot showed transcription of the target gene, with a temporal expression pattern similar to those from previous work. Western blot showed assembly of various immune complexes from monomer to hexamer. Partial purification of scEDIII-PIGS was also attempted to demonstrate the feasibility of yeast system for immune complex vaccine development. Approximately 1 mg of scEDIII-PIGS can be produced from 1 l culture. Conclusion This work demonstrated for the first time that various immunocomplex structures of our target protein could be efficiently produced in S. cerevisiae for future application in developing oral and injectable vaccines against various pathogens.

Expression of virus-like particles (VLPs) of foot-and-mouth disease virus (FMDV) using Saccharomyces cerevisiae

“…S. cerevisiae 2805 ( MATa pep4::HIS3 prb1-Δcan1 GAL2 his3 ura3-52 ) was the yeast strain used for heterologous expression. Yeast and bacterial strains were cultured, as previously described (So et al 2021 ).…”

“…Yeast cells transformed with empty pYEGPD-TER vector were constructed as a negative control. The presence of the transformed recombinant plasmid was assessed by colony PCR analysis and E. coli back-transformation (So et al 2021 ).

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“…S. cerevisiae has been considered as a generally recognized as safe (GRAS) organism, which generates its recombinant proteins and development processes readily applicable with no further consideration. Furthermore, due to the strong adjuvant property of yeast derivatives, yeast expression offers an attractive system for vaccine production and development (Bal et al 2018a , b ; Cen et al 2021 ; Gao et al 2021 ; Goh et al 2021 ; Kim et al 2010 ; Seif et al 2016 ; So et al 2021 ). Some of our recent studies have demonstrated that the dengue viral epitope expressed by this yeast expression induced a well-established immune response to dengue virus (Bal et al 2018a , b ; Kim et al 2010 ; Nguyen et al 2013 , 2015 ; Seif et al 2016 ).…”

Metabolic engineering of Saccharomyces cerevisiae for the biosynthesis of a fungal pigment from the phytopathogenic fungus Cladosporium phlei

“…The S. cerevisiae strain was transformed as previously reported [ 30 – 32 , 35 , 36 ]. The transformed cells were selected on the selection medium, and the presence of the transforming plasmid was confirmed by colony PCR or back-transformation of E. coli with DNA prepared from the putative transformants [ 31 ].…”

“…Subsequently, 250 uL of the primary inoculum was inoculated into 5 mL of YEPD medium and incubated for 16 h at 30 °C with continuous agitation (200 rpm). This 5 mL culture was transferred into a 300-mL Erlenmeyer flask containing 40 mL of selection medium at 20 °C, 25 °C, and 30 °C with continuous agitation (200 rpm), after which the cells were harvested and examined for phleichrome expression [30][31][32].…”