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Rapid optical imaging of EGF receptor expression with a single-chain antibody SNAP-tag fusion protein

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

The epidermal growth factor receptor (EGFR) is overexpressed in several types of cancer and its inhibition can effectively inhibit tumour progression. The purpose of this study was to design an EGFR-specific imaging probe that combines efficient tumour targeting with rapid systemic clearance to facilitate non-invasive assessment of EGFR expression.

Methods

Genetic fusion of a single-chain antibody fragment with the SNAP-tag produced a 48-kDa antibody derivative that can be covalently and site-specifically labelled with substrates containing 0 6-benzylguanine. The EGFR-specific single-chain variable fragment (scFv) fusion protein 425(scFv)SNAP was labelled with the near infrared (NIR) dye BG-747, and its accumulation, specificity and kinetics were monitored using NIR fluorescence imaging in a subcutaneous pancreatic carcinoma xenograft model.

Results

The 425(scFv)SNAP fusion protein accumulates rapidly and specifically at the tumour site. Its small size allows efficient renal clearance and a high tumour to background ratio (TBR) of 33.2 ± 6.3 (n = 4) 10 h after injection. Binding of the labelled antibody was efficiently competed with a 20-fold excess of unlabelled probe, resulting in an average TBR of 6 ± 1.35 (n = 4), which is similar to that obtained with a non-tumour-specific probe (5.44 ± 1.92, n = 4). When compared with a full-length antibody against EGFR (cetuximab), 425(scFv)SNAP-747 showed significantly higher TBRs and complete clearance 72 h post-injection.

Conclusion

The 425(scFv)SNAP fusion protein combines rapid and specific targeting of EGFR-positive tumours with a versatile and robust labelling technique that facilitates the attachment of fluorophores for use in optical imaging. The same approach could be used to couple a chelating agent for use in nuclear imaging.

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Abbreviations

hAGT:

Human O 6-alkyl-DNA alkyltransferase

NIR:

Near infrared

scFv:

Single-chain variable fragment

EGFR:

Epidermal growth factor receptor 1

BG:

Benzylguanine

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Acknowledgments

We would like to thank Dr. Richard Twyman for the critical reading of the manuscript and Agnieszka Dreier (Institute for Neuropathology, University Hospital, RWTH Aachen University) for supplying the cetuximab antibody.

Conflicts of interest

None.

Author information

Authors and Affiliations

  1. Fraunhofer Institute for Molecular Biology and Applied Ecology, 52074, Aachen, Germany

    Florian Kampmeier, Judith Niesen, Alexander Koers, Markus Ribbert, Rainer Fischer, Stefan Barth & Theo Thepen

  2. Covalys Biosciences AG, 4108, Witterswil, Switzerland

    Andreas Brecht

  3. Institute for Molecular Biotechnology, RWTH Aachen University, 52074, Aachen, Germany

    Rainer Fischer

  4. Department of Experimental Molecular Imaging, Medical Faculty, RWTH Aachen University, 52074, Aachen, Germany

    Fabian Kießling

  5. Department of Experimental Medicine and Immunotherapy, Helmholtz Institute for Biomedical Engineering, RWTH Aachen University, 52074, Aachen, Germany

    Stefan Barth

Authors
  1. Florian Kampmeier
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  2. Judith Niesen
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  3. Alexander Koers
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  4. Markus Ribbert
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  7. Fabian Kießling
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  9. Theo Thepen
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Corresponding author

Correspondence to Theo Thepen.

Additional information

Stefan Barth and Theo Thepen contributed equally to this work.

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Kampmeier, F., Niesen, J., Koers, A. et al. Rapid optical imaging of EGF receptor expression with a single-chain antibody SNAP-tag fusion protein. Eur J Nucl Med Mol Imaging 37, 1926–1934 (2010). https://doi.org/10.1007/s00259-010-1482-5

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  • DOI: https://doi.org/10.1007/s00259-010-1482-5

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