S1 domain of the porcine epidemic diarrhea virus spike protein as a vaccine antigen

Makadiya, Niraj; Brownlie, Robert; van den Hurk, Jan; Berube, Nathalie; Allan, Brenda; Gerdts, Volker; Zakhartchouk, Alexander

The S1 ORF plus C-terminal his 10 tag (codon optimized for mammalian expression) together with a proceeding Kozak sequence, was cloned downstream of a human CMV promoter plus intron, contained within an inhouse episomal vector; elements downstream of the S1 ORF included a woodchuck hepatitis post-transcriptional regulatory element and bovine growth hormone polyadenylation site (DNA sequence of constructs are available upon request). Constructs were transfected into HEK293T cells using Turbofect (ThermoFisher Scientific) according to the manufacturer's instructions. Cells stably maintaining the episomal constructs were selected by puromycin. For protein production, stably transfected cells were grown in SFM4HEK293 medium (ThermoFisher Scientific) with shaking at 37 0 C and 5 % CO 2 . Supernatant was harvested and processed for purification.