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European Journal of Nuclear Medicine and Molecular Imaging

Published in:

01-10-2010 | Original Article

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

Authors: Florian Kampmeier, Judith Niesen, Alexander Koers, Markus Ribbert, Andreas Brecht, Rainer Fischer, Fabian Kießling, Stefan Barth, Theo Thepen

Published in: European Journal of Nuclear Medicine and Molecular Imaging | Issue 10/2010

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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Title: Rapid optical imaging of EGF receptor expression with a single-chain antibody SNAP-tag fusion protein
Authors: Florian Kampmeier
Judith Niesen
Alexander Koers
Markus Ribbert
Andreas Brecht
Rainer Fischer
Fabian Kießling
Stefan Barth
Theo Thepen
Publication date: 01-10-2010
Publisher: Springer-Verlag
Published in: European Journal of Nuclear Medicine and Molecular Imaging / Issue 10/2010
Print ISSN: 1619-7070
Electronic ISSN: 1619-7089
DOI: https://doi.org/10.1007/s00259-010-1482-5

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

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

Abstract

Purpose

Methods

Results

Conclusion

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusion

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