Screening HIV-1 fusion inhibitors based on capillary electrophoresis head-end microreactor targeting to the core structure of gp41.
Screening HIV-1 fusion inhibitors based on capillary electrophoresis head-end microreactor targeting to the core structure of gp41.
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DOI：
10.1016/j.jpba.2015.12.021
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发表时间：
2016-02
影响因子：
3.4
通讯作者：
Lihong Liu;Xiaoying Xu;Yanhui Liu;Xuanxuan Zhang;Lin Li;Zhimin Jia
中科院分区：
文献类型：
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作者：
Lihong Liu;Xiaoying Xu;Yanhui Liu;Xuanxuan Zhang;Lin Li;Zhimin Jia
本文设计了一种基于毛细管电泳（CE）的电泳介导微量分析（EMMA）的微反应器，用于筛选与N端七肽重复序列（NHR，N36）结合的HIV-1抑制剂。首先，将测试样品塞加载到填充有缓冲溶液的毛细管中，然后是N36肽溶液，并且两种溶液通过扩散同时混合。然后，施加电压，样品分子通过N36肽区。活性化合物联合收割机与N36结合，导致活性化合物的峰高损失。对100多种中药提取物进行了筛选，得到了地龙（Pheretima aspergillum，E. Perrier）（L5）被鉴定为具有有效的抑制活性。结果表明，L5能显著抑制HIV-1 JR-FL假型病毒感染，其半数有效浓度（EC 50）约为32.1 ± 1.2 μg/mL，半数细胞毒性浓度（CC 50）为146.9 ± 4.4 μg/mL，表明L5对U87-CD 4-CCR 5细胞具有较低的体外细胞毒性。新方法简单快速，不含抗体，不需要繁琐的过程。
In this paper, we design a microreactor based on electrophoretically mediated microanalysis (EMMA) with capillary electrophoresis (CE) for screening HIV-1 inhibitors that bind to the N-terminal heptad repeat (NHR, N36) region. Initially, a test sample plug is loaded into a capillary filled with buffer solution followed by N36 peptide solution, and the two solutions simultaneously mix by diffusion. Then, voltage is applied, and the sample molecules pass through the N36 peptide zone. The active compounds combine with N36, leading to a loss in the peak height of the active compound. More than 100 traditional Chinese medicine extracts (TCME) were screened, and an extract ofPheretima aspergillum(E. Perrier) (L5) was identified as having potent inhibitory activity. The results showed that L5 could significantly inhibit the HIV-1JR-FLpseudotyped virus infection; the 50% effective concentration (EC50) of L5 was approximately 32.1 ± 1.2 μg/mL, and the 50% cytotoxicity concentration (CC50) value of L5 was 146.9 ± 4.4 μg/mL, suggesting that L5 had lowin vitrocytotoxicity on U87-CD4-CCR5 cells. The new method is simple and rapid, is free of antibodies, and does not require tedious processes.