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目的 制备针对朊蛋白的特异性抗体。方法 以初步纯化的原核细胞表达的GST PrP融合蛋白免疫家兔 ,制备朊蛋白特异的抗体。结果 以GST PrP s为抗原 ,用酶联免疫吸咐试验(ELISA)检测免疫家兔血清中的朊病毒抗体效价最高可达至 12 80 0 0。Westernbolt检测显示所制备的抗体不仅可以与体外表达的全长及不同C 末端缺失的人朊蛋白反应 ,而且能够识别人、鼠脑组织中内源性朊蛋白。结论 利用GST PrP融合蛋白可有效地剌激免疫动物产生PrP特异性抗体 ,所制备的抗血清可试用于朊蛋白的检测。
Objective To prepare specific antibodies against prion protein. Methods Rabbit was immunized with GST-PrP fusion protein expressed in primary purified prokaryotic cells to prepare prion-specific antibodies. Results Using GST PrP as antigen, the titer of prion antibody in rabbit serum was up to 12 80 0 0 by enzyme-linked immunosorbent assay (ELISA). Westernbolt assay showed that the prepared antibody not only reacted with human prion protein with full length and different C terminus expressed in vitro, but also recognized endogenous prion protein in human and mouse brain tissue. Conclusion The GST PrP fusion protein can effectively stimulate the animals to produce PrP-specific antibodies. The prepared antisera can be used for the detection of prion protein.