用微流控器件捕获肿瘤特有的细胞外囊泡
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By LabMedica International staff writers Posted on 17 Apr 2018 |
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EVHB芯片表面上捕捉到的病人肿瘤排出的细胞外囊泡（红色）（图片蒙Shannon Stott博士惠赐）。
日前开发成功的一款微流控器件能捕获胶质母细胞瘤产生的细胞外囊泡，特异性高，所需血样量少，适用于儿童患者。
器件内部的微流控通道含有多种抗体的组合，这些抗体是胶质母细胞瘤产生的细胞外囊泡上的分子特有的，说明囊泡经过通道时被捕获。
美国马萨诸塞州查尔斯顿麻省总医院的科学家及其同事收集了13名脑癌患者的血样，研究中还包括6名健康献血者。微流控器件由8通道鲱鱼骨结构构成，用标准光刻法制造，并试验了不同的策略，以找出在微流控器件表面上捕捉抗体的最佳构型。
用新西兰基督堂市Izon Science公司的可调电阻脉冲感应(TRPS) qNano仪器量化分离的细胞外囊泡(EV)。用固定的链霉亲和素涂层磁性粒子分离EV。这款环烯烃共聚物(COC)微流控器件可以直接显像捕获的EV。用美国马萨诸塞州皮博迪市卡尔蔡司公司(www.zeiss.com)的配备63倍蔡司Plan-APOCHROMAT油浸物镜的LSM510共焦显微镜获取显微图像。分离并量化RNA。还进行了数字聚合酶链反应、为RNA测序准备文库以及RNA测序分析。
科研小组反映，这套灵敏的微流控分析平台(EVHB-Chip)能在三小时内进行肿瘤特异的EV-RNA分离。他们用EVHB-Chip检测肿瘤EV达到了94%的特异性，检测限为每微升100 EV，肿瘤RNA的浓度是其它方法的10倍。随后这种方法可以释放捕获的肿瘤EV，便于下游测定和功能研究。他们通过处理多形性胶质母细胞瘤(GBM)患者的血清和血浆样本检测表皮生长因子受体III型变体(EGFRvIII)的信使RNA (mRNA)。
首席研究员、医学助理教授Shannon L. Stott博士说：“我们的器件能够从血流中数十亿个细胞外囊泡中分拣肿瘤特异的细胞外囊泡，这样可以为这种癌症和其它难治的癌症开发急需的诊断工具和监护工具。”该研究的论文发表于2018年1月12日《自然》杂志《通信》分册的网络版。
麻省总医院>>> www.massgeneral.org
Izon Science >>> www.izon.com
卡尔蔡司>>> www.zeiss.com
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