Heterogeneous immunoassays using magnetic beads on a digital microfluidic platform.
Heterogeneous immunoassays using magnetic beads on a digital microfluidic platform.
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DOI：
10.1039/b807855f
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发表时间：
2008-12
期刊：
影响因子：
6.1
通讯作者：
Pamula VK
中科院分区：
文献类型：
--
作者：
Sista RS;Eckhardt AE;Srinivasan V;Pollack MG;Palanki S;Pamula VK
本文提出了一种基于磁珠有效处理的数字微流控平台，用于进行异质三明治免疫测定。这种方法是基于使用电润湿操作样品和试剂的离散液滴，而不需要液滴自由横向移动的通道。因此，基于液滴的磁珠操作不会受到通道堵塞的影响。在数字微流控平台上进行免疫分析需要以下基本操作：头部吸引，头部洗涤，头部保留和头部重悬。对磁场强度、拉力、位置和缓冲成分等参数进行了研究。用永磁体固定磁珠，用电润湿法分离多余的上清，证明了磁珠的稀释洗涤。在对上清液进行7776倍稀释洗涤后，几乎100%的珠粒保留。通过在平台上的五个电极上运输带有磁珠的液滴，并利用液滴内部的流动模式来重悬浮磁珠，实现了磁珠的有效重悬浮。将所有磁珠滴滴操作整合在一起，生成人胰岛素和白细胞介素-6 （IL-6）夹心异质免疫测定的标准曲线，每次测定的总时间为7分钟。
A digital microfluidic platform for performing heterogeneous sandwich immunoassays based on efficient handling of magnetic beads is presented in this paper. This approach is based on manipulation of discrete droplets of samples and reagents using electrowetting without the need for channels where the droplets are free to move laterally. Droplet-based manipulation of magnetic beads therefore does not suffer from clogging of channels. Immunoassays on a digital microfluidic platform require the following basic operations: bead attraction, bead washing, bead retention, and bead resuspension. Several parameters such as magnetic field strength, pull force, position, and buffer composition were studied for effective bead operations. Dilution-based washing of magnetic beads was demonstrated by immobilizing the magnetic beads using a permanent magnet and splitting the excess supernatant using electrowetting. Almost 100% bead retention was achieved after 7776 fold dilution-based washing of the supernatant. Efficient resuspension of magnetic beads was achieved by transporting a droplet with magnetic beads across five electrodes on the platform and exploiting the flow patterns within the droplet to resuspend the beads. All the magnetic-bead droplet operations were integrated together to generate standard curves for sandwich heterogeneous immunoassays on Human Insulin and Interleukin-6 (IL-6) with a total time to result of seven minutes for each assay.
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影响因子：
6.1
作者：
Moon, Hyejin;Wheeler, Aaron R.;Kim, Chang-Jin CJ''
通讯作者：
Kim, Chang-Jin CJ''
影响因子：
2.7
作者：
Cho, SK;Moon, HJ;Kim, CJ
通讯作者：
Kim, CJ
影响因子：
8.4
作者：
Shikida, M;Takayanagi, K;Sato, K
通讯作者：
Sato, K
影响因子：
7.4
作者：
Sato, K;Tokeshi, M;Kitamori, T
通讯作者：
Kitamori, T
影响因子：
2.9
作者：
Moorthy, J;Mensing, GA;Beebe, DJ
通讯作者：
Beebe, DJ