Rojo Núria, Ercilla Guadalupe, Haro Isabel
Department of Peptide & Protein Chemistry, IIQAB-CSIC, Jordi Girona 18-26, 08034 Barcelona, Spain. ihvqpp@ iiqab.csic.es
Curr Protein Pept Sci. 2003 Aug;4(4):291-8. doi: 10.2174/1389203033487135.
In the present study, new putative epitopes located in structural (E2) and non-structural (NS3) proteins of GBV-C/HGV were identified by computer-aided prediction of antigenicity and synthesized in solid-phase, following an Fmoc/tBut strategy, for their use in immunoassays. The corresponding synthetic peptides were used as antigens in ELISA assays and in real-time biospecific interaction measurements. This last approach allowed direct detection of GBV-C/HGV-specific antibodies in human sera. Good correlations were obtained between the biospecific interaction analysis and the ELISA. To verify the performance of these new assays in comparison to the existing recombinant E2 protein commercial test, antibodies to synthetic peptides were searched for in different panels of serum samples. The main conclusion of this work is the usefulness of E2 peptides in the detection of antibodies. Moreover, the NS3 peptide could be exploited to improve the sensitivity of the currently available test. Our results offer a new approach to develop new diagnostic peptide based biosensors for serodiagnosis of GBV-C/HGV infection.
在本研究中，通过计算机辅助预测抗原性，在GBV-C/HGV的结构蛋白（E2）和非结构蛋白（NS3）中鉴定出新的假定表位，并按照Fmoc/tBut策略在固相合成，用于免疫测定。相应的合成肽用作ELISA检测和实时生物特异性相互作用测量的抗原。后一种方法能够直接检测人血清中的GBV-C/HGV特异性抗体。生物特异性相互作用分析与ELISA之间获得了良好的相关性。为了验证这些新检测方法与现有的重组E2蛋白商业检测相比的性能，在不同血清样本组中寻找针对合成肽的抗体。这项工作的主要结论是E2肽在抗体检测中的有用性。此外，NS3肽可用于提高现有检测的灵敏度。我们的结果为开发用于GBV-C/HGV感染血清学诊断的新型诊断肽基生物传感器提供了一种新方法。
