人类PrP合成肽抗原构建及应用

摘要

目的为提高Prion病诊断率，解决PrP抗原来源不足，人工合成人类PrP多肽用于抗体制备。方法应用固相合成法合成15个氨基酸残基的人类PrP多肽，并利用碳二亚胺法将其与牛血清白蛋白偶连制成免疫原，应用于动物免疫及多克隆抗体的制备。结果偶连后PrP多肽具有免疫原性，并获得了抗PrP多克隆抗体。结论人类PrP合成肽抗原的获得和应用，可以部分替代天然抗原，为进一步单克隆抗体制备，以及Prion病的深入研究提供了有利条件。%Aim In order to improver the diagnostic rate of Prion disease, solve the lacking sources of nature PrP antigen. ,synthesize the human PrP peptide, prepare the antibody to PrP peptide further. Method A synthetic peptide with the sequence identical to the 15 residues of human PrP, as described by Kretzschmar[1], was synthesized by the solid-phase method. The synthetic peptide was coupled to bovine serum albumin(BSA)by the method of EDCI〔2〕. The polypeptide combined with BSA was used as antigen to immunize the rabbit and detected by immunomethod. Result The PrP polypeptide combined with BSA obtained immunogenicity and anti-PrP synthetic peptide antiserum was successfully obtained. Conclusion The preparation and application of human PrP synthetic peptide can substitute for nature PrP antigen partly. It has laid a foundation for further preparation of monoclonal antibody to PrP and the study of Prion disease.