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Updated: May 19, 2026

Covalent Immobilization of Proteins for the Single Molecule Force Spectroscopy
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Evaluation of Complex Formation between Aminated Polyrotaxane and Proteins Using FRET.

Hiroya Tsukahara1, Toru Taharabaru1, Kosei Utatsu1

  • 1Graduate School of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Chuo-ku, Kumamoto 862-0973, Japan.

Chemical & Pharmaceutical Bulletin
|May 17, 2026
PubMed
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Researchers developed fluorescent aminated polyrotaxane (amino-PRX) carriers to track intracellular delivery. This visualization method confirmed complex formation and distribution within cells, demonstrating potential for antibody delivery.

Area of Science:

  • Biomaterials Science
  • Cellular Biology
  • Drug Delivery Systems

Background:

  • Aminated polyrotaxane (amino-PRX) is an efficient intracellular carrier for biopharmaceuticals.
  • The detailed intracellular behavior of amino-PRX and its complexes, especially for antibody delivery, requires further investigation.

Purpose of the Study:

  • To visualize the intracellular distribution of amino-PRX carriers and their complexes using fluorescence.
  • To examine the intracellular behavior of amino-PRX complexes and assess their potential for antibody delivery.

Main Methods:

  • Preparation of fluorescently capped amino-PRX (DET-PRX-FAM) using α-cyclodextrin and polyethylene glycol.
  • Utilizing Förster resonance energy transfer (FRET) to observe complex formation between the carrier and model proteins (phycocyanin, IgG).
Keywords:
Förster resonance energy transferantibodycarrierintracellular deliverypolyrotaxane

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  • Confocal microscopy in HeLa cells to visualize carrier, drug, and FRET fluorescence.

    • Main Results:

    • DET-PRX-FAM (2 kDa) showed lower cytotoxicity than the 20 kDa variant in HeLa cells.
    • FRET-derived fluorescence confirmed complex formation between DET-PRX-FAM (2 kDa) and model proteins.
    • Intracellular visualization of carrier, protein, and FRET signals demonstrated successful complex delivery and distribution.

    Conclusions:

    • Fluorescently capped amino-PRX enables visualization of intracellular carrier and complex dynamics.
    • The study validates amino-PRX as a potential carrier for intracellular antibody delivery.
    • This approach enhances understanding of carrier-mediated intracellular transport mechanisms.