Department of Chemistry and Institute for Biophysical Dynamics, The University of Chicago, 929 East 57th Street, Chicago, Illinois 60637, USA.
Anal Chem. 2010 Apr 15;82(8):3276-82. doi: 10.1021/ac100044c.
This article 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 and (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 approximately 13 pM. A total of 48 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.
本文描述了一种基于 SlipChip 的方法,用于使用多个纳升级体积的样品进行基于珠的异质免疫分析。作为分析 chemistrode 输出的潜在设备,该平台的性能使用低浓度的生物分子进行了测试。在 SlipChip 中进行免疫分析的两种策略进行了测试:(1) 单向滑动方法,将含有样品的孔与一系列预先加载有试剂的孔组合;(2) 来回滑动方法,通过重新加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。SlipChip 采用亲水表面的内部孔和疏水表面的 SlipChip 表面制造,以增强在浅孔中填充、转移和保持水溶液的能力。纳米图案化用于增加 SlipChip 表面的疏水性。含有捕获抗体的磁性珠在孔之间高效转移并通过连续稀释进行洗涤。胰岛素酶联免疫测定显示出约 13 pM 的检测限。总共分析了 48 个纳升级的液滴,包括芯片上的校准。SlipChip 的设计灵活,可以适应其他类型的免疫分析,包括异质和同质免疫分析。这项工作为使用 SlipChip 进行小体积免疫分析开辟了可能性,适用于一系列可能的应用,包括分析 chemistrode 中的插头、检测单细胞中的分子以及诊断监测。