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Related Experiment Videos

Protein labelling via deoxyribonucleic acid hybridization

D J Hnatowich1, G Mardirossian, M Rusckowski

  • 1Department of Nuclear Medicine, University of Massachusetts Medical Center, Worcester 01655, USA.

Nuclear Medicine Communications
|January 1, 1996
PubMed
Summary

This study introduces a new method for radiolabelling proteins using DNA hybridization. Modified antibodies showed stable labeling with technetium-99m, with properties influenced by DNA type.

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Area of Science:

  • Bioconjugation Chemistry
  • Radiopharmaceutical Science
  • Molecular Imaging

Background:

  • Antibody-based imaging and therapy requires efficient and stable radiolabelling methods.
  • Current radiolabelling techniques can face challenges with stability and specific activity.
  • DNA hybridization offers a potential platform for novel radiolabelling strategies.

Purpose of the Study:

  • To develop and characterize a novel method for radiolabelling antibodies and proteins using single-stranded DNA conjugation and hybridization.
  • To evaluate the efficiency, stability, and biodistribution of radiolabelled antibodies prepared by this method.
  • To investigate the influence of DNA backbone chemistry (diester vs. thioate) on radiolabel properties.

Main Methods:

  • Covalent conjugation of single-stranded DNA (ssDNA) to antibodies.

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  • Hybridization of ssDNA-conjugated antibodies with complementary ssDNA labelled with technetium-99m (99Tc(m)) or indium-111 (111In).
  • In vitro stability assays in saline and serum.
  • In vivo biodistribution studies in normal mice.

    • Main Results:

    • Achieved 100% antibody labelling efficiency in approximately 1 hour at room temperature.
    • Obtained high specific activities of up to 30 microCi/microgram for 99Tc(m)-labelled IgG.
    • Diester-DNA labelled antibodies showed label degradation in serum, while thioate-DNA labelled antibodies exhibited non-specific binding to serum proteins.
    • Biodistribution revealed rapid renal clearance for diester-DNA conjugates and predominant liver deposition for thioate-DNA conjugates.

    Conclusions:

    • Proteins can be readily radiolabelled using DNA hybridization following ssDNA modification.
    • The choice of DNA backbone significantly impacts the stability and biodistribution of the radiolabel.
    • This DNA-based approach offers a versatile platform for developing novel radiopharmaceuticals.