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利用合成肽进行丙型肝炎病毒(HCV)的线性抗原表位分析
Screening of Linear Antigen Epitopes Using Synthetic Peptides Derived from Structural and Non-structural Proteins of HCV
【摘要】 首先利用固相多肽合成技术对HCV多蛋白中可能含有优势抗原表位的14个肽段进行了化学合成,并用合成肽作包被抗原进行间接ELLSA试验来分析它们的抗原性,结果表明,所有合成肽中除NS3区的P7、P8和NS5区的P13无抗原性外,其余肽段均具有抗原活性,其中C区的P1,NS4区和P9和NS5区的P12三个肽段的抗原性较好,与相应的重组蛋白C22,C100和NS5A比较,它们之间的抗原性比较接近,随后选择抗原性较好的几个肽段合成了含八分支的多抗原肽(MAP)MP1、MP2、MP3、MP10和嵌合多肽CP15等工程肽抗原,前者不仅保持了相应单体肽的抗原性,而且与抗体反应的敏感性有显著提高;后者含双表位改善了常规合成肽抗原表位单一的缺陷,为研究HCV工程肽抗原进行了有益的尝试。
【Abstract】 Fourteen peptides derived from structural and non-structural proteins of HCV and harboring putative antigen epitopes were synthesized with solid phase peptide synthesis(SPPS) technology firstly.The results of indirect ELISA using these peptides as coating antigens indicated that all these peptides ,except P7,PS and P13,showed antigenic reactivity in 56 anti-HCV positive serum which were verified by Abbott anti-HCV EIA(Ⅱ).Among them,PI,P9 and P12 were shown to be highly immunoreactive.On the basis of the peptides with dominant linear antigen epitope ,engineering peptides including octa-branched peptide(MP) and chimeric peptide(CP) were then synthesized,immunoassay results showed that branch peptides can improve distinctly the coating efficiency and sensitivity of antigen-antibody reaction.
【Key words】 HCV; Peptide and engineering peptide; ELISA; Epitope screening;
- 【文献出处】 生物化学杂志 , 编辑部邮箱 ,1996年04期
- 【分类号】R373.21
- 【被引频次】3
- 【下载频次】127