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捕获与释放细胞分选:T细胞从抗体修饰微电极上的电化学解吸

Catch and release cell sorting: electrochemical desorption of T-cells from antibody-modified microelectrodes.

作者信息

Zhu He, Yan Jun, Revzin Alexander

机构信息

Department of Biomedical Engineering, University of California, Davis, 451 East Health Sciences Street #2619, Davis, CA 95616, United States.

出版信息

Colloids Surf B Biointerfaces. 2008 Jul 15;64(2):260-8. doi: 10.1016/j.colsurfb.2008.02.010. Epub 2008 Mar 10.

Abstract

The development of integrated microsystems capable of interrogation, characterization and sorting of mammalian cells is highly significant for further advancement of point-of-care diagnostics and drug discovery fields. The present study sought to design a novel strategy for releasing antibody-bound cells through electrochemical disruption of the underlying antibody (Ab) layer. A microsystem for selective capture and release of cells consisted of an array of individually addressable gold microelectrodes fabricated on a glass substrate. Poly(ethylene glycol) (PEG) hydrogel photolithography was employed to make the glass regions non-fouling, thus, ensuring selective localization of proteins and cells on the microelectrodes. The gold surfaces were decorated with anti-CD4 Ab molecules using standard alkanethiol self-assembly and carbodiimide coupling approaches. The Ab-functionalized electrodes selectively captured model T-lymphocytes (Molt-3 cells) expressing CD4 antigen while minimal cell adhesion was observed on PEG hydrogel-modified glass substrates. Importantly, application of a reductive potential (-1.2V vs. Ag/AgCl reference electrode) resulted in release of surface-bound T-cells from the electrode surface. Cyclic voltammetry and fluorescence microscopy were employed to verify that the detachment of captured T-cells was indeed due to the electrochemical disruption of the underlying alkanethiol-Ab layer. In the future, the cell sorting approach described here may be combined with microfluidic delivery to enable Ab-mediated capture of T-lymphocytes or other cell types followed by release of select cells for downstream gene expression studies or re-cultivation.

摘要

能够对哺乳动物细胞进行检测、表征和分选的集成微系统的开发,对于即时诊断和药物发现领域的进一步发展具有极其重要的意义。本研究旨在设计一种新策略,通过对底层抗体(Ab)层进行电化学破坏来释放与抗体结合的细胞。一种用于选择性捕获和释放细胞的微系统由在玻璃基板上制造的一系列可单独寻址的金微电极组成。采用聚(乙二醇)(PEG)水凝胶光刻技术使玻璃区域具有抗污性,从而确保蛋白质和细胞在微电极上的选择性定位。使用标准的烷硫醇自组装和碳二亚胺偶联方法,在金表面装饰抗CD4 Ab分子。经Ab功能化的电极选择性地捕获表达CD4抗原的模型T淋巴细胞(Molt-3细胞),而在PEG水凝胶修饰的玻璃基板上观察到极少的细胞粘附。重要的是,施加还原电位(相对于Ag/AgCl参比电极,为-1.2V)会导致表面结合的T细胞从电极表面释放。采用循环伏安法和荧光显微镜来验证捕获的T细胞的脱离确实是由于底层烷硫醇-Ab层的电化学破坏。未来,这里描述的细胞分选方法可与微流体输送相结合,以实现Ab介导的T淋巴细胞或其他细胞类型的捕获,随后释放选定的细胞用于下游基因表达研究或再培养。

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