【摘要】 目的探讨妊娠相关血浆蛋白A(PAPP-A)的抗原表位及抗PAPP-A抗体制备方法。方法把PAPP-A的CDS序列翻译成氨基酸序列,利用互联网分析蛋白的疏水性和信号肽区段,利用DNAstar软件分析二级结构、表面可及性、亲水性、抗原指数以及柔韧性,综合分析抗原表位。通过分子生物学技术制备重组PAPP-A,进而免疫新西兰家兔制备抗PAPP-A抗体。结果 PAPP-A抗原性较强,抗原优势表位位于23～94、1 110～1 131氨基酸区段;PAPP-A重组蛋白以及抗PAPP-A抗体制备成功。结论本研究方法建立了PAPP-A的高灵敏免疫检测体系,能进一步提高PAPP-A的临床应用价值。更多还原

【Abstract】 Objective To analyze epitopes of PAPP-A and prepare pregnancy-associated plasma protein A(PAPP-A) antibody.Methods CDS region of PAPP-A was translated into amino acid sequencing form;then hydrophobicity and signal peptide of PAPP-A were analyzed by Internet;the secondary structure,surface probability,hydrophilicity,antigen index and flexible region were all analyzed by DNAstar;then the antigenic epitopes were analyzed by the above means.Recombinant PAPP-A antigen was made by a series of molecular biology techniques and then it was injected into rabbits to make PAP-A antibody.Results Antigenicity of PAPP-A was strong,the region was located in 23-94,1110-1131 region of the amino acid sequencing.The recombinant PAPP-A antigen and its antibody were managed to prepare.Conclusion The high sensitive immunoassay of PAPP-A was established in this study,which can furthur improve the clinical application value of PAPP-A.更多还原