分支肽的合成与蛋白芯片的制作

【作者】 闫婕；

【导师】 陈五岭；

【作者基本信息】 西北大学 ， 微生物学， 2004， 硕士

【摘要】 本文采用固相合成方法人工合成了人巨细胞病毒(HCMV)抗原表位的分支多肽，并用斑点金免疫渗滤试验进行了HCMV抗体的检测。研究结果表明： 构建了分支多肽的支架结构，采用分支状多聚赖氨酸和线形多聚赖氨酸两种，以两种不同的支架结构合成了八分支肽。 将支架结构上的氨基酸作为合成的结合位点，按照常规方法依次进行后续氨基酸的连接，由于空间位阻效应，后续氨基酸的连接困难度增加，八分支肽中的残缺肽较多，由此方法合成的分支肽反应活性较低。 将支架结构上的氨基酸作为合成的结合位点，将已合成并纯化好的线形肽作为待连接的短肽，连接到支架上，从而得到残缺肽极少的分支状八肽，效果较好。 以线形多聚赖氨酸作为支架所合成的八分支肽的反应活性高于以分支状多聚赖氨酸作为支架所合成的八分支肽。 所合成的分支多肽在蛋白芯片的硝酸纤维素膜上具有良好的结合活性和抗原活性，可用于人巨细胞病毒抗体的快速临床检测。更多还原

【Abstract】 The author synthesized the multiple antigenic peptides with solid phase method. The MAPs were made of the human cytomlegalovirus’s antigen epitope. And the DIGFA method was applied to detect the antibody against HCMV. The result shows:The MAPs were constructed by two kinds of scaffolding: linear lysine-tree scaffolding and branchy lysine-tree scaffolding.One of the synthetic methods of the MAP is to connect subsequent amino acid one by one with the lysine belonging to the lysine-tree scaffolding. Because of the steric hinderance effect, synthesis becomes more and more difficult. And for so many crippled peptides in the MAP, the reactivity of it is not so good.The MAP connecting pured peptide to the lysine-tree scaffolding has less crippled peptides and better reactivity.The MAPs linked to different lysine-tree scaffolding have different immunogenicity. The reactivity of the MAP made by linear lysine-tree scaffolding is better than the reactivity of the MAP made by branchy lysine-tree scaffolding.The protein chip made by coating the MAP on the nitrocellulose membrane could be developed to be a new rapid clinical detecting method because of its high binding activity and the antigenic activity of the HCMV.更多还原