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通过介电泳操作快速形成细胞-颗粒复合物,用于检测表面抗原。

Rapid formation of cell-particle complexes via dielectrophoretic manipulation for the detection of surface antigens.

机构信息

Graduate School of Material Science, University of Hyogo,3-2-1 Kouto, Kamigori, Ako, Hyogo 678-1297, Japan.

Graduate School of Material Science, University of Hyogo,3-2-1 Kouto, Kamigori, Ako, Hyogo 678-1297, Japan; JST-CREST, 5, Sanbancho, Chiyoda, Tokyo 102-0075, Japan.

出版信息

Biosens Bioelectron. 2014 Nov 15;61:215-21. doi: 10.1016/j.bios.2014.05.019. Epub 2014 May 21.

DOI:10.1016/j.bios.2014.05.019
PMID:24892783
Abstract

A rapid and simple method for the fabrication of the island patterns with particles and cells was applied to detect the presence of specific antigens on the cell surface. An upper interdigitated microband array (IDA) electrode was mounted on a lower substrate with the same design to fabricate a microfluidic-channel device for dielectrophoretic manipulation. The electrode grid structure was fabricated by rotating the upper template IDA by 90° relative to the lower IDA. A suspension of anti-CD33 modified particles and HL-60 cells was introduced into the channel. An AC electrical signal (typically 20 V peak-to-peak, 100 kHz) was then applied to the bands of the upper and lower IDAs, resulting in the formation of island patterns at the intersections with low electric fields. Immunoreactions between the antibodies immobilized on the accumulated particles and the CD33 present on the surface of the cells led to the formation of complexes comprising corresponding antigen-antibody pairs. Non-specific pairs accumulated at the intersection, which did not form complexes, were then dispersed after removal of the applied field. The time required for the detection of the formation/dispersion of the complexes is as short as 6 min in the present procedure. Furthermore, this novel cell binding assay does not require pretreatment such as target labeling or washing of the unbound cells.

摘要

一种快速简便的颗粒和细胞岛图案制作方法被应用于检测细胞表面特定抗原的存在。在上层交错微带阵列 (IDA) 电极上安装具有相同设计的下层基底,以制造用于介电泳操作的微流控通道装置。电极网格结构通过将上层模板 IDA 相对于下层 IDA 旋转 90°来制造。将抗 CD33 修饰颗粒和 HL-60 细胞的悬浮液引入通道。然后将交流电信号(通常为 20 V 峰峰值,100 kHz)施加到上下 IDA 的带,导致在电场较低的交点处形成岛图案。固定在积累颗粒上的抗体与细胞表面上存在的 CD33 之间的免疫反应导致形成包含相应抗原-抗体对的复合物。非特异性对在未形成复合物的交点处积累,然后在去除施加的场后分散。在本程序中,检测复合物的形成/分散所需的时间短至 6 分钟。此外,这种新型细胞结合测定不需要目标标记或未结合细胞的洗涤等预处理。

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