Establishment of a Madin–Darby bovine kidney cell line expressing anchorless bovine prion protein

Abstract

Enzyme-linked immunosorbent assay (ELISA) performed using extensively purified bacterially expressed bovine prion protein (PrP) shows decreased cross-reactivity. We generated a transduced Madin–Darby bovine kidney (MDBK) cell line continuously expressing glycosylphosphatidylinositol (GPI)-anchorless bovine PrP (designated as MDBK ∆GPI protein) by using a lentiviral expression system. The present study also described the method for purifying bovine PrP through sequential culturing without the need for complex purification protocol. Our results showed that the purified bovine PrP could be used as an immunogen for developing anti-PrP monoclonal antibodies. Together, our results suggest that the new GPI-anchorless bovine PrP and its purification method can be used for performing basic studies for employing a cell-based approach.