biostudies-literatureLee EJMinistry of Trade, Industry and EnergyNational Research Foundation of KoreaKorea Research Institute of Bioscience and Biotechnologye2506050https://www.ebi.ac.uk/biostudies/studies/S-EPMC12463560biostudies-literatureUnknown35The <i>N</i>-terminal S1 subunit of the spike protein (S1 protein) of porcine epidemic diarrhea virus (PEDV) is recognized as a potential diagnostic antigen for detecting anti-PEDV immunoglobulin A (IgA) levels in colostrum, a key indicator for assessing passive immunity against PEDV infection. Given the advantages of producing Fc-fusion proteins in mammalian cells, it is important to investigate how the addition of the Fc region affects the diagnostic performance of the PEDV-S1 protein. In this study, we successfully produced full-length PEDV-S1 protein fused with an Fc region (PEDV-S1-Fc protein) via transient gene expression in recombinant Chinese hamster ovary (rCHO) cells adapted to serum-free suspension culture. The resulting expression showed significantly higher volumetric productivity than previously reported. The aglycosylated form of PEDV-S1-Fc protein was also generated using PNGase F treatment of the purified protein, as well as tunicamycin treatment to inhibit glycosylation during cell culture. Notably, neither the addition of the Fc region nor the removal of <i>N</i>-glycans markedly affected the diagnostic function, as demonstrated by indirect ELISA using PEDV-positive and PEDV-negative colostrum samples. Moreover, the diagnostic performance of the PEDV-S1-Fc protein was further validated using total IgA purified from PEDV-positive colostrum. In summary, the full-length Fc-fused PEDV-S1 protein produced in rCHO cell culture exhibited high productivity and purity, making it a promising antigenic candidate for indirect ELISA-based detection of anti-PEDV IgA in colostrum.Journal of microbiology and biotechnologyProduction of Recombinant S1 Protein of Porcine Epidemic Diarrhea Virus in Recombinant CHO Cells for Application in Indirect ELISA.PMC12463560RS-2024-00443405KGM1262511Kim SKim THWon HLee EJHeo NYKoo SJKim HSKim YGRyu SHfalseProduction of Recombinant S1 Protein of Porcine Epidemic Diarrhea Virus in Recombinant CHO Cells for Application in Indirect ELISA.The <i>N</i>-terminal S1 subunit of the spike protein (S1 protein) of porcine epidemic diarrhea virus (PEDV) is recognized as a potential diagnostic antigen for detecting anti-PEDV immunoglobulin A (IgA) levels in colostrum, a key indicator for assessing passive immunity against PEDV infection. Given the advantages of producing Fc-fusion proteins in mammalian cells, it is important to investigate how the addition of the Fc region affects the diagnostic performance of the PEDV-S1 protein. In this study, we successfully produced full-length PEDV-S1 protein fused with an Fc region (PEDV-S1-Fc protein) via transient gene expression in recombinant Chinese hamster ovary (rCHO) cells adapted to serum-free suspension culture. The resulting expression showed significantly higher volumetric productivity than previously reported. The aglycosylated form of PEDV-S1-Fc protein was also generated using PNGase F treatment of the purified protein, as well as tunicamycin treatment to inhibit glycosylation during cell culture. Notably, neither the addition of the Fc region nor the removal of <i>N</i>-glycans markedly affected the diagnostic function, as demonstrated by indirect ELISA using PEDV-positive and PEDV-negative colostrum samples. Moreover, the diagnostic performance of the PEDV-S1-Fc protein was further validated using total IgA purified from PEDV-positive colostrum. In summary, the full-length Fc-fused PEDV-S1 protein produced in rCHO cell culture exhibited high productivity and purity, making it a promising antigenic candidate for indirect ELISA-based detection of anti-PEDV IgA in colostrum.2025-01-01T00:00:00Z2025 Sep2026-06-03T20:09:30.543Z2026-05-31T03:07:29.637ZS-EPMC124635604096790910.4014/jmb.2506.06050