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A dual fluorochrome probe for imaging proteases.

Moritz F Kircher1, Ralph Weissleder, Lee Josephson

  • 1Center for Molecular Imaging Research, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129, USA.

Bioconjugate Chemistry
|March 18, 2004
PubMed
Summary

This study introduces a novel dual fluorochrome probe for noninvasive in vivo protease activity imaging. The probe uses distinct near-infrared fluorescence signals to accurately detect protease activity, independent of lesion depth or size.

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Area of Science:

  • Biomedical Imaging
  • Nanotechnology
  • Enzyme Activity Assays

Background:

  • Near-infrared fluorescence (NIRF) optical probes enable noninvasive assessment of enzyme activity and pathology.
  • Current methods for enzyme activity assessment can be limited in specificity and depth penetration.

Purpose of the Study:

  • To develop and characterize a novel dual fluorochrome enzyme-activatable probe for in vivo protease activity imaging.
  • To establish a dual-wavelength imaging method for protease activity that is independent of lesion size and depth.

Main Methods:

  • A dual fluorochrome probe was synthesized using amine functional cross-linked iron oxide (CLIO) nanoparticles.
  • One NIRF fluorochrome (Cy7) was attached in a protease-resistant manner, serving as an internal standard.

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  • A second NIRF fluorochrome (Cy5.5) was attached via a protease-sensitive polyarginine peptide spacer.

    • Main Results:

    • Protease activity increased the ratio of Cy5.5 to Cy7 fluorescence both in vitro and in vivo.
    • Absolute fluorescence values correlated with lesion size and depth.
    • The fluorescence ratio remained constant, independent of lesion size and depth, indicating accurate protease activity measurement.

    Conclusions:

    • The dual fluorochrome probe and dual wavelength imaging method offer a novel approach for in vivo protease activity imaging.
    • This technology provides a reliable method for assessing protease activity in various pathologies.
    • The probe's design ensures accurate measurements irrespective of lesion characteristics, enhancing diagnostic potential.