SlipChip for immunoassays in nanoliter volumes.
SlipChip for immunoassays in nanoliter volumes.
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DOI：
10.1021/ac100044c
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发表时间：
2010-04-15
影响因子：
7.4
通讯作者：
Ismagilov, Rustem F.
中科院分区：
文献类型：
--
作者：
Liu, Weishan;Chen, Delai;Du, Wenbin;Nichols, Kevin P.;Ismagilov, Rustem F.
本文描述了一种基于滑动芯片的方法，用于对多个纳升体积的样品进行基于微珠的异质免疫测定。作为分析化学电极输出的潜在装置，使用低浓度的生物分子测试了该平台的性能。测试了在滑动芯片中进行免疫测定的两种策略：1）单向滑动方法，将含有样品的孔与一系列预装试剂的孔组合起来； 2)往复滑动法，通过重新装载和滑动含有试剂的孔，将一系列试剂引入含有样品的孔中。滑动芯片在孔内部具有亲水表面，在滑动芯片表面具有疏水表面，以增强浅孔中的填充、转移和维持水溶液。纳米图案化用于增加滑动芯片表面的疏水性。含有捕获抗体的磁珠在孔之间有效转移并通过连续稀释进行洗涤。胰岛素免疫酶测定显示检测限约为 13 pM。并行分析了 48 个纳升体积的液滴，包括片上校准。滑动芯片的设计非常灵活，可以适应其他类型的免疫测定，包括异质和同质的。这项工作确立了小体积使用基于滑动芯片的免疫测定的可能性，用于一系列可能的应用，包括化学电极插头的分析、单细胞分子的检测和诊断监测。
This paper describes a SlipChip-based approach to perform bead-based heterogeneous immunoassays with multiple nanoliter-volume samples. As a potential device to analyze the output of the chemistrode, the performance of this platform was tested using low concentrations of biomolecules. Two strategies to perform the immunoassay in the SlipChip were tested: 1) a unidirectional slipping method to combine the well containing a sample with a series of wells preloaded with reagents; 2) a back-and-forth slipping method to introduce a series of reagents to a well containing the sample by reloading and slipping the well containing the reagent. The SlipChips were fabricated with hydrophilic surfaces on the interior of the wells and with hydrophobic surfaces on the face of the SlipChip to enhance filling, transferring, and maintaining aqueous solutions in shallow wells. Nanopatterning was used to increase the hydrophobic nature of the SlipChip surface. Magnetic beads containing the capture antibody were efficiently transferred between wells and washed by serial dilution. An insulin immunoenzymatic assay showed a detection of limit of ~13 pM. Forty eight droplets of nanoliter volume were analyzed in parallel, including an on-chip calibration. The design of the SlipChip is flexible to accommodate other types of immunoassays, both heterogeneous and homogeneous. This work establishes the possibility of using SlipChip-based immunoassays in small volumes for a range of possible applications, including analysis of plugs from a chemistrode, detection of molecules from single cells, and diagnostic monitoring.
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