Kim Hyo-Jin, Roh In-Soon, Park Hoo-Chang, Ahn Su Bi, Suh Tae-Young, Park Kyung-Je, Kang Hae-Eun, Sohn Hyun-Joo
OIE Reference Laboratory for CWD, Foreign Animal Disease Research Division, Animal and Plant Quarantine Agency, Gimcheon, Gyeongsangbukdo 39660, Korea.
J Vet Med Sci. 2018 Jun 6;80(6):909-912. doi: 10.1292/jvms.17-0521. Epub 2018 Apr 4.
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.
使用广泛纯化的细菌表达牛朊病毒蛋白（PrP）进行的酶联免疫吸附测定（ELISA）显示交叉反应性降低。我们通过慢病毒表达系统生成了一种转导的马-达二氏牛肾（MDBK）细胞系，该细胞系持续表达糖基磷脂酰肌醇（GPI）无锚定牛PrP（命名为MDBK ∆GPI蛋白）。本研究还描述了通过连续培养纯化牛PrP的方法，无需复杂的纯化方案。我们的结果表明，纯化的牛PrP可用作开发抗PrP单克隆抗体的免疫原。总之，我们的结果表明，新的GPI无锚定牛PrP及其纯化方法可用于基于细胞方法的基础研究。
