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    Use of photoimmunoconjugates to characterize ABCB1 in cancer cells

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    Author
    Liang, Barry J.
    Lusvarghi, Sabrina
    Ambudkar, Suresh V.
    Huang, Huang Chiao
    Date
    2021-07-26
    Journal
    Nanophotonics
    Publisher
    De Gruyter Open Ltd
    Type
    Article
    
    Metadata
    Show full item record
    See at
    https://doi.org/10.1515/nanoph-2021-0252
    Abstract
    Accurate detection of ATP-binding cassette drug transporter ABCB1 expression is imperative for precise identification of drug-resistant tumors. Existing detection methods fail to provide the necessary molecular details regarding the functional state of the transporter. Photoimmunoconjugates are a unique class of antibody–dye conjugates for molecular diagnosis and therapeutic treatment. However, conjugating hydrophobic photosensitizers to hydrophilic antibodies is quite challenging. Here, we devise a photoimmunoconjugate that combines a clinically approved benzoporphyrin derivative (BPD) photosensitizer and the conformational-sensitive UIC2 monoclonal antibody to target functionally active human ABCB1 (i.e., ABCB1 in the inward-open conformation). We show that PEGylation of UIC2 enhances the BPD conjugation efficiency and reduces the amount of non-covalently conjugated BPD molecules by 17%. Size exclusion chromatography effectively separates the different molecular weight species found in the UIC2–BPD sample. The binding of UIC2–BPD to ABCB1 was demonstrated in lipidic nanodiscs and ABCB1-overexpressing triple negative breast cancer (TNBC) cells. UIC2–BPD was found to retain the conformation sensitivity of UIC2, as the addition of ABCB1 modulators increases the antibody reactivity in vitro. Thus, the inherent fluorescence capability of BPD can be used to label ABCB1-overexpressing TNBC cells using UIC2–BPD. Our findings provide insight into conjugation of hydrophobic photosensitizers to conformation-sensitive antibodies to target proteins expressed on the surface of cancer cells.
    Sponsors
    National Science Foundation
    Keyword
    ABCB1
    benzoporphyrin derivative
    photodynamic therapy
    photoimmunoconjugate
    UIC2
    Identifier to cite or link to this item
    http://hdl.handle.net/10713/16944
    ae974a485f413a2113503eed53cd6c53
    10.1515/nanoph-2021-0252
    Scopus Count
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