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

Anal Chim Acta. 2020 Jun 15:1116:53-61. doi: 10.1016/j.aca.2020.03.048. Epub 2020 Apr 8.

Abstract

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)6]4-/3- and also improved the reversibility of redox. These improvements are due to the interaction between [Fe(CN)6]4-/3- 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.

Keywords: Cytosensing; EIS; Electron-transfer peptide; K562 cell; Marrow-origin peptide.

MeSH terms

  • Amino Acid Sequence
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Collagen / chemistry
  • Dielectric Spectroscopy / instrumentation
  • Dielectric Spectroscopy / methods*
  • Electrodes
  • Humans
  • Immobilized Proteins / chemistry*
  • Immobilized Proteins / metabolism
  • K562 Cells
  • Neoplasm Proteins / metabolism
  • Oligopeptides / chemistry*
  • Oligopeptides / metabolism
  • Receptors, Cell Surface / metabolism

Substances

  • Immobilized Proteins
  • Neoplasm Proteins
  • Oligopeptides
  • Receptors, Cell Surface
  • phenylalanyl-arginyl-prolyl-arginyl-isoleucyl-methionyl-threonyl-proline
  • Collagen