Assessment of synthetic peptides for hepatitis A diagnosis using biosensor technology
Assessment of synthetic peptides for hepatitis A diagnosis using biosensor technology
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DOI：
10.1016/s0022-1759(00)00295-7
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发表时间：
2000-12-01
影响因子：
2.2
通讯作者：
Haro, I
中科院分区：
文献类型：
--
作者：
Gómara, MJ;Ercilla, G;Haro, I
在目前的工作中，我们证明了商业生物传感器仪器（BIACORE 1000，Biacore AB，乌普萨拉）的应用，用于检测抗体的甲型肝炎病毒（HAV）在人血清样品中使用线性和分支合成肽相关的VP 3衣壳蛋白的HAV。我们还通过圆二色性（CD）研究了合成肽的构象，以分析可能影响其被抗体识别的构建体的二级结构的变化。线性和二聚体VP 3（110-121）多抗原肽（MAP）是最敏感的，适用于使用BIACORE对HAV感染患者血清进行血清学研究。通过胺基固定四聚体MAP显然不能保持肽表位的活性构象，因为与线性和二聚体肽相比，它导致较低的抗体结合。CD分析显示，由于分子间聚集，四聚体MAP构建体倾向于采用β-折叠结构，这限制了表位可及性。我们的结果表明，使用合成肽的生物特异性相互作用分析技术在急性甲型肝炎诊断中的价值。(C)2000 Elsevier Science B. V.保留所有权利。
In the present work we demonstrate the application of a commercial biosensor instrument (BIACORE 1000, Biacore AB, Uppsala) for the detection of antibodies against the hepatitis A virus (HAV) in human serum samples using linear and branched synthetic peptides related to the VP3 capsid protein of HAV. We also studied the conformation of the synthetic peptides by circular dichroism (CD) in order to analyse the changes in secondary structure of the constructs that could influence their recognition by antibodies. Linear and dimeric VP3(110-121) multiple antigen peptides (MAP) were the most sensitive and appropriate for serological studies of serum from HAV infected patients using BIACORE. Immobilization of tetrameric MAPs via amine groups apparently failed to preserve the active conformation of the peptide epitope since it led to lower antibody binding compared to linear and dimeric peptides. The CD analysis showed that the tetrameric MAP constructs tend to adopt a beta -sheet structure due to intermolecular aggregation, which limits epitope accessibility. Our results demonstrate the Value of biospecific interaction analysis technology using synthetic peptides for the diagnosis of acute hepatitis A. (C) 2000 Elsevier Science B.V. All rights reserved.