6533b852fe1ef96bd12aae98
RESEARCH PRODUCT
Site-Specific Dual-Labeling of a VHH with a Chelator and a Photosensitizer for Nuclear Imaging and Targeted Photodynamic Therapy of EGFR-Positive Tumors
Franck DenatMartin GotthardtEmma RenardEstel Collado CampsSanne A. M. Van LithAnnemarie KipColine CanovasMark RijpkemaVictor Goncalvessubject
Cancer ResearchFluorescence-lifetime imaging microscopyBiodistribution[SDV.IB.IMA]Life Sciences [q-bio]/Bioengineering/Imagingmedia_common.quotation_subjectmedicine.medical_treatmentPhotodynamic therapyvariable domain of heavy chain only antibodies (VHH); site-specific conjugation; dual-labeling; nuclear imaging; photodynamic therapy[SDV.CAN]Life Sciences [q-bio]/Cancer[CHIM.THER]Chemical Sciences/Medicinal Chemistrylcsh:RC254-282Article030218 nuclear medicine & medical imaging03 medical and health sciencesTumours of the digestive tract Radboud Institute for Health Sciences [Radboudumc 14]0302 clinical medicineAll institutes and research themes of the Radboud University Medical CenterIn vivoduallabelingmedicineTumours of the digestive tract Radboud Institute for Molecular Life Sciences [Radboudumc 14]PhotosensitizerInternalizationmedia_commonnuclear imagingChemistrysite-specific conjugationlcsh:Neoplasms. Tumors. Oncology. Including cancer and carcinogens3. Good healthOncologyphotodynamic therapy030220 oncology & carcinogenesisUrological cancers Radboud Institute for Health Sciences [Radboudumc 15]dual-labelingBiophysicsvariable domain of heavy chain only antibodies (VHH)A431 cellsEx vivodescription
Simple Summary Variable domains of heavy chain only antibodies are small proteins that can be used for tumor imaging and therapy upon conjugation of functional groups. As frequently used random conjugation techniques can decrease binding to the target of interest, site-specific conjugation of these functional groups is preferred. Here, we optimized site-specific conjugation of both a chelator for binding of a radiometal and a photosensitizer to epidermal growth factor receptor (EGFR) binding VHH 7D12. We characterized this dual-labeled VHH for nuclear imaging and targeted photodynamic therapy of EGFR-expressing tumors. Abstract Variable domains of heavy chain only antibodies (VHHs) are valuable agents for application in tumor theranostics upon conjugation to both a diagnostic probe and a therapeutic compound. Here, we optimized site-specific conjugation of the chelator DTPA and the photosensitizer IRDye700DX to anti-epidermal growth factor receptor (EGFR) VHH 7D12, for applications in nuclear imaging and photodynamic therapy. 7D12 was site-specifically equipped with bimodal probe DTPA-tetrazine-IRDye700DX using the dichlorotetrazine conjugation platform. Binding, internalization and light-induced toxicity of DTPA-IRDye700DX-7D12 were determined using EGFR-overexpressing A431 cells. Finally, ex vivo biodistribution of DTPA-IRDye700DX-7D12 in A431 tumor-bearing mice was performed, and tumor homing was visualized with SPECT and fluorescence imaging. DTPA-IRDye700DX-7D12 was retrieved with a protein recovery of 43%, and a degree of labeling of 0.56. Spectral properties of the IRDye700DX were retained upon conjugation. 111In-labeled DTPA-IRDye700DX-7D12 bound specifically to A431 cells, and they were effectively killed upon illumination. DTPA-IRDye700DX-7D12 homed to A431 xenografts in vivo, and this could be visualized with both SPECT and fluorescence imaging. In conclusion, the dichlorotetrazine platform offers a feasible method for site-specific dual-labeling of VHH 7D12, retaining binding affinity and therapeutic efficacy. The flexibility of the described approach makes it easy to vary the nature of the probes for other combinations of diagnostic and therapeutic compounds.
year | journal | country | edition | language |
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2021-01-23 | Cancers |