Towards a peptide-based suspension array for the detection of pestivirus antibodies in swine
Towards a peptide-based suspension array for the detection of pestivirus antibodies in swine
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DOI：
10.1016/j.jviromet.2016.04.022
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发表时间：
2016-09-01
影响因子：
3.1
通讯作者：
Loeffen, Willie L. A.
中科院分区：
文献类型：
--
作者：
van der Wal, Fimme J.;Jelsma, Tinka;Loeffen, Willie L. A.
猪瘟是一种高度传染性和致死性的猪瘟。诊断脑脊液的血清学试验不仅需要检测CSFV抗体，还需要将这些抗体与其他鼠疫病毒抗体区分开来。为了探索基于多肽的特异性血清学的可能性，用代表CSFV E2蛋白线性表位(TAVSPTTLR)的各种合成多肽来测试基于多肽的悬浮阵列用于检测猪瘟病毒抗体的可行性。结果表明，N端生物素化的多肽可以结合到亲和素偶联的小球上，并在检测相应的单抗WH303中发挥作用。有迹象表明，表位和生物素之间的间隔区长度对肽-抗体相互作用的效率有影响。建立了一种能够探测猪血清中抗体的方法，其中血清中和亲和素的阻断和使用空白对照珠进行标准化是至关重要的。以一组具有多种鼠疫病毒抗体的猪血清为例，论证了基于多肽的悬浮阵列用于特异性检测猪血清中的鼠疫病毒抗体的概念的正确性。(C)2016年提交人。爱思唯尔出版公司(Elsevier B.V.)
Classical swine fever (CSF) is a highly contagious and lethal disease in swine. Serological tests for the diagnosis of CSF need not only to detect antibodies against CSFV, but also need to differentiate these from antibodies against other pestiviruses. To investigate the possibilities of specific peptide-based serology, various synthetic peptides that represent a well-described linear epitope of the CSFV E2 protein (TAVSPTTLR) were used to test the viability of a peptide-based suspension array for the detection of antibodies against pestiviruses in swine.The results show that N-terminally biotinylated peptides can bind to avidin conjugated beads, and function in detection of the corresponding monoclonal antibody WH303. There are indications that the length of the spacer between epitope and biotin affect the efficiency of the peptide-antibody interaction. A protocol was established that enables probing for antibodies in porcine sera, where neutravidin-blocking of serum and the use of empty control beads for normalization was crucial. With a set of porcine sera with antibodies against various pestiviruses, the proof of concept of a peptide-based suspension array for specific detection of antibodies against pestiviruses in porcine sera was demonstrated. (C) 2016 The Authors. Published by Elsevier B.V.