Computationally Designed Peptides for Zika Virus Detection: An Incremental Construction Approach
Computationally Designed Peptides for Zika Virus Detection: An Incremental Construction Approach
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DOI：
10.3390/biom9090498
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发表时间：
2019-09-01
期刊：
影响因子：
5.5
通讯作者：
Daunert, Sylvia
中科院分区：
文献类型：
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作者：
Mascini, Marcello;Dikici, Emre;Daunert, Sylvia
在这里，与目前生产的抗寨卡病毒抗体不同，我们提出了一种半组合虚拟策略，选择短肽作为仿生抗体/结合剂来检测完整的寨卡病毒(ZIKV)颗粒。该虚拟方法基于通过最大化ZIKV和登革病毒(DENV)包膜蛋白糖基化位点的氨基酸基序之间的区分来生成四、五、六和七肽文库的不同对接循环。然后合成了8个多肽，每个长度2个(四、五、六和七肽)，并用直接酶联免疫吸附试验(ELISA)与完整的ZIKV颗粒进行比较。作为参考，我们使用了一种成熟的抗ZIKV抗体，抗体4G2。三种基于多肽的检测方法具有良好的检测限，动态范围从10(5)拷贝/毫升的完整ZIKV颗粒开始；这比其他多肽或抗体低一个数量级。在完整病毒粒子的浓度为10(6)拷贝/毫升时，这三种多肽与DENV-1、-2和-3三种血清型病毒有轻微的交叉反应，但当包被浓度增加到病毒的10(7)拷贝/毫升时，DENV和ZIKV之间的区别就消失了。在血清和尿液稀释度分别为1：10和1：1的两种生物基质存在下检测多肽的敏感性。对于血清或尿液中寨卡病毒的检测，检出限降低了大约一个数量级，尽管对于三种多肽中的两种，仍有一个明显的分析信号，从急性感染时的寨卡病毒浓度10(6)拷贝/毫升开始。
Herein, and in contrast to current production of anti-Zika virus antibodies, we propose a semi-combinatorial virtual strategy to select short peptides as biomimetic antibodies/binding agents for the detection of intact Zika virus (ZIKV) particles. The virtual approach was based on generating different docking cycles of tetra, penta, hexa, and heptapeptide libraries by maximizing the discrimination between the amino acid motif in the ZIKV and dengue virus (DENV) envelope protein glycosylation site. Eight peptides, two for each length (tetra, penta, hexa, and heptapeptide) were then synthesized and tested vs. intact ZIKV particles by using a direct enzyme linked immunosorbent assay (ELISA). As a reference, we employed a well-established anti-ZIKV antibody, the antibody 4G2. Three peptide-based assays had good detection limits with dynamic range starting from 10(5) copies/mL of intact ZIKV particles; this was one order magnitude lower than the other peptides or antibodies. These three peptides showed slight cross-reactivity against the three serotypes of DENV (DENV-1, -2, and -3) at a concentration of 10(6) copies/mL of intact virus particles, but the discrimination between the DENV and ZIKV was lost when the coating concentration was increased to 10(7) copies/mL of the virus. The sensitivity of the peptides was tested in the presence of two biological matrices, serum and urine diluted 1:10 and 1:1, respectively. The detection limits decreased about one order of magnitude for ZIKV detection in serum or urine, albeit still having for two of the three peptides tested a distinct analytical signal starting from 10(6) copies/mL, the concentration of ZIKV in acute infection.