Enabling Indium Channels for Mass Cytometry by Using Reinforced Cyclam-Based Chelating Polylysine

Bioconjug Chem. 2020 Sep 16;31(9):2103-2115. doi: 10.1021/acs.bioconjchem.0c00267. Epub 2020 Aug 12.

Abstract

The synthesis of a polylysine polymer functionalized with the previously reported astonishingly inert [In(cb-te2pa)]+ chelate was performed. A biotin end group allowed the conjugation to biotinylated beads by the intermediary of a fluorescein isothiocyanate/neutravidin receptor. High quality imaging mass cytometry trials, based on 115In detection were performed to highlight the behavior of the material. Anti-CD20 antibody was labeled by the so-obtained In(III)-modified polylysine using the biotin/neutravidin interaction. Ramos (CD20[+]) and HL-60 (CD20[-]) cell lines were costained with the In(III)-modified bioconjugate by finding the best staining conditions. Both immunofluorescence microscopy (IF-M) and mass cytometry analyses confirmed the specific binding of anti-CD20 onto Ramos cells. CyTOF histograms constructed on the 115In detection allowed us to define and to separate, with a good signal-to-noise ratio, two populations (Ramos and HL-60). The inertness of In(III)-MCP-NAv over a three-month storage period was proved by performing new functionality tests involving Jurkat cells (CD20[-]) and multiparametric trials involving the 115In channel. The results ensure a promising future use of the previously announced [In(cb-te2pa)]+ complex-based polymers for mass cytometry.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / chemistry
  • Antigens, CD20 / analysis*
  • Biotinylation
  • Cell Line
  • Chelating Agents / chemistry
  • Flow Cytometry / methods
  • Fluorescein-5-isothiocyanate / analogs & derivatives*
  • Fluorescent Dyes / chemistry
  • HL-60 Cells
  • Heterocyclic Compounds / chemistry*
  • Humans
  • Immunoconjugates / chemistry*
  • Indium / chemistry*
  • Jurkat Cells
  • Models, Molecular
  • Polylysine / chemistry*

Substances

  • Antibodies, Monoclonal
  • Antigens, CD20
  • Chelating Agents
  • Fluorescent Dyes
  • Heterocyclic Compounds
  • Immunoconjugates
  • Indium
  • Polylysine
  • cyclam
  • Fluorescein-5-isothiocyanate