Fabrication of a cell-recognition/electron-transfer/cross-linker, peptide-immobilized electrode for the sensing of K562 cells.

We designed an electrode that has the ability to sense a target cell. This new electrode is intended for use in cell recognition via electron-transfer and cross-linker peptide immobilization. Myelopeptide-4 (MP-4:FRPRIMTP) is a marrow-origin peptide that interacts with receptors of the human leukemia cell line (K562 cells), and allows their differentiation. The YYYYC electron-transfer peptide improves the electron-transfer accessibility from an electroactive compound to an electrode. Oligoalanine plays the role of a cross-linker that immobilizes a peptide series (Ac-FRPRIMTPYYYYCAAAAA) to collagen, which then allows it to be cast onto an electrode. Use of the electrode with a peptide increased the peak currents of [Fe(CN)] and also improved the reversibility of redox. These improvements are due to the interaction between [Fe(CN)] and the peptide. When electrochemical impedance spectroscopy (EIS) measurements were carried out using a collagen/peptide probe-immobilized electrode, the electron transfer resisitance was lower than that without the peptide. The detection of K562 cells was based on an increase in resistance, because MP-4 was bound to the receptors on the cell surface. The responses were linear and ranged in number from 27 to 2,000 cells/mLwith a detection limit of 8 cells/mL. Recoveries of 50 and 1,000 cells/mL in human serum were accomplished at rates of 98 and 101%, respectively. Consequently, the proposed procedure is a powerful new concept for cytosensing.