Zinc oxide nanorod based immunosensing platform for the determination of human leukemic cells

Talanta. 2019 Aug 1:200:378-386. doi: 10.1016/j.talanta.2019.03.064. Epub 2019 Mar 16.

Abstract

Zinc oxide (ZnO) based nanostructures owing unique physical properties - high photoluminescence, biocompatibility and other characteristics, therefore, they attract attention as building blocks suitable for biosensor development. In this research as a target we have used human leukemic cell line IM9 (IM9). IM9 was derived from the patient with a multiple myeloma and expressed cluster of differentiation proteins СD19 on the surface of 85-95% here investigated cancer cells. As a control sample healthy human's peripheral blood mononuclear cells (PBMC) were used and the expression of CD19 protein was found only in 5-9% of these cells. Two types of antibodies labeled by fluorescein isothiocyanate (FITC) were used for the labeling of human leukemic cells: FITC-conjugated mouse antibodies against Human CD19 protein (anti-CD19-FITC*) and FITC-conjugated mouse antibodies against Human IgG1 protein (anti-IgG1-FITC*). In order to demonstrate the applicability of zinc oxide nanorods (ZnO-NRs) based platforms three types of ZnO-NRs-based structures were investigated: (i) ZnO-NRs modified by anti-CD19-FITC*; (ii) ZnO-NRs modified by IM9 cells, which were pre-incubated with anti-CD19-FITC*; (iii) ZnO-NRs modified by PBMC cells, which were pre-incubated with anti-CD19-FITC*. It was demonstrated that IM9 cells after specific interaction with anti-CD19-FITC* bind to ZnO-NRs (ZnO-NRs/IM9 +anti-CD19-FITC*) and photoluminescence based signal significantly increase in comparison with that observed in control samples, which contained PBMC cells incubated with anti-CD19-FITC* (ZnO-NRs/PBMC+anti-CD19-FITC*). The photoluminescence results are in good correlation with the data obtained by flow cytometry. This study illustrate that ZnO-NRs exhibit a photoluminescence signal suitable for the determination of anti-CD19-FITC* labeled IM9 cell line at concentrations - from 10 till 500 cells adsorbed per 1 mm2 of ZnO-NRs platform.

Keywords: Flow cytometry; Human lymphocytes; Monoclonal antibody; Photoluminescence; ZnO-nanorods; В-lymphoblast cells.

MeSH terms

  • B-Lymphocytes / pathology*
  • Biomarkers / blood
  • Cell Separation*
  • Cells, Cultured
  • Flow Cytometry
  • Humans
  • Immunoassay / methods*
  • Leukemia / pathology*
  • Nanotubes / chemistry*
  • Zinc Oxide / chemical synthesis
  • Zinc Oxide / chemistry*

Substances

  • Biomarkers
  • Zinc Oxide