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

Laura Grenier1, Maryline Beyler1, Taunia Closson2

  • 1Univ Brest, UMR-CNRS 6521 CEMCA, 6 Avenue Victor le Gorgeu, 29200 BREST, France.

Bioconjugate Chemistry
|June 23, 2020
PubMed
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Researchers developed a new indium(III)-modified polylysine polymer for mass cytometry. This bioconjugate specifically targets CD20-positive cells, enabling precise cell population identification and separation in complex biological samples.

Area of Science:

  • Bioconjugation Chemistry
  • Mass Cytometry
  • Immunophenotyping

Background:

  • Development of novel bioconjugates for advanced cellular analysis.
  • Need for stable and specific labeling agents in high-resolution cytometry.
  • Inertness of indium(III) chelates for biological applications.

Purpose of the Study:

  • To synthesize and characterize a polylysine polymer functionalized with an inert indium(III) chelate.
  • To evaluate the utility of this bioconjugate for antibody labeling and cell targeting in mass cytometry.
  • To assess the specificity and stability of the indium(III)-modified polymer for cell population discrimination.

Main Methods:

  • Synthesis of biotinylated polylysine functionalized with [In(cb-te2pa)]+ chelate.
  • Conjugation to biotinylated beads via fluorescein isothiocyanate/neutravidin.

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  • Labeling of anti-CD20 antibody and subsequent mass cytometry (CyTOF) analysis of cell lines (Ramos, HL-60, Jurkat).

    • Main Results:

    • Successful synthesis and characterization of the indium(III)-modified polylysine bioconjugate.
    • Specific binding of anti-CD20 to Ramos (CD20+) cells confirmed by immunofluorescence microscopy and mass cytometry.
    • Clear separation of CD20+ and CD20- cell populations (Ramos vs. HL-60) using 115In detection with good signal-to-noise ratio.
    • Demonstrated inertness and stability of the bioconjugate over a three-month storage period.

    Conclusions:

    • The developed indium(III)-modified polylysine polymer is a stable and effective tool for antibody labeling in mass cytometry.
    • The bioconjugate enables specific targeting and discrimination of cell populations based on biomarker expression.
    • This technology holds significant promise for future applications in high-parameter single-cell analysis and diagnostics.