Microfluidic passive permeability assay using nanoliter droplet interface lipid bilayers
Microfluidic passive permeability assay using nanoliter droplet interface lipid bilayers
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DOI：
10.1039/c3an01314f
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发表时间：
2013-01-01
期刊：
影响因子：
4.2
通讯作者：
Schmidt, Jacob J.
中科院分区：
文献类型：
--
作者：
Nisisako, Takasi;Portonovo, Shiva A.;Schmidt, Jacob J.
膜通透性测定在评价药物跨生物膜转运活性方面起着重要作用。然而，在传统的平行人工膜透过率分析(PAMPA)中，所使用的膜模型在物理和化学上与生物膜不同。在这里，我们描述了一种使用液滴界面双层(DIBS)的微流控被动渗透性分析方法。在微流体网络中，在含有磷脂的交叉流动油中交替形成纳升大小的给体和受体水滴。随后，通过疏水的伪多孔性侧壁选择性地去除油，诱导脂单分子层接触，在供体和受体液滴之间形成排列的平面滴状。荧光素从供体到受体液滴的渗透被荧光计量法测量。根据测量数据和一个简单的扩散模型，我们计算了在pH值为7.5、6.4和5.4时，供体液滴的有效渗透系数分别为5.1×10(-6)cm S(-1)、60.0×10(-6)cm S(-1)和87.6×10(-6)cm S(-1)。获得了特定的单阴离子和中性荧光素的本征渗透率。我们还用紫外光谱测定了咖啡因在10分钟内的渗透率，得到咖啡因的渗透率为20.8×10(-6)cm S(-1)。由于溶液体积小，测量时间短，能够测量范围广泛的化合物，该设备作为高通量药物渗透性分析的平台具有相当大的潜力。
Membrane permeability assays play an important role in assessing drug transport activities across biological membranes. However, in conventional parallel artificial membrane permeability assays (PAMPA), the membrane model used is dissimilar to biological membranes physically and chemically. Here, we describe a microfluidic passive permeability assay using droplet interface bilayers (DIBs). In a microfluidic network, nanoliter-sized donor and acceptor aqueous droplets are alternately formed in cross-flowing oil containing phospholipids. Subsequently, selective removal of oil through hydrophobic pseudo-porous sidewalls induces the contact of the lipid monolayers, creating arrayed planar DIBs between the donor and acceptor droplets. Permeation of fluorescein from the donor to the acceptor droplets was fluorometrically measured. From the measured data and a simple diffusion model we calculated the effective permeabilities of 5.1 x 10(-6) cm s(-1), 60.0 x 10(-6) cm s(-1), and 87.6 x 10(-6) cm s(-1) with donor droplets at pH values of 7.5, 6.4 and 5.4, respectively. The intrinsic permeabilities of specific monoanionic and neutral fluorescein species were obtained similarly. We also measured the permeation of caffeine in 10 min using UV microspectroscopy, obtaining a permeability of 20.8 x 10(-6) cm s(-1). With the small solution volumes, short measurement time, and ability to measure a wide range of compounds, this device has considerable potential as a platform for high-throughput drug permeability assays.