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

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 (−1.2 V 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.