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Development and preclinical characterisation of 99mTc-labelled Affibody molecules with reduced renal uptake

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Abstract

Purpose

Affibody molecules are low molecular weight proteins (7 kDa), which can be selected to bind to tumour-associated target proteins with subnanomolar affinity. Because of rapid tumour localisation and clearance from nonspecific compartments, Affibody molecules are promising tracers for molecular imaging. Earlier, 99mTc-labelled Affibody molecules demonstrated specific targeting of tumour xenografts. However, the biodistribution was suboptimal either because of hepatobiliary excretion or high renal uptake of the radioactivity. The goal of this study was to optimise the biodistribution of Affibody molecules by chelator engineering.

Materials and methods

Anti-HER2 ZHER2:342 Affibody molecules, carrying the mercaptoacetyl-glutamyl-seryl-glutamyl (maESE), mercaptoacetyl-glutamyl-glutamyl-seryl (maEES) and mercaptoacetyl-seryl-glutamyl-glutamyl (maSEE) chelators, were prepared by peptide synthesis and labelled with 99mTc. The tumour-targeting capacity of these conjugates was compared with each other and with the best previously available conjugate, 99mTc-maEEE-ZHER2:342, in nude mice bearing SKOV-3 xenografts. The tumour-targeting capacity of the most promising conjugate, 99mTc-maESE-ZHER2:342, was compared with radioiodinated ZHER2:342.

Results

All novel conjugates demonstrated successful tumour targeting and a low degree of hepatobiliary excretion. The renal uptakes of serine-containing conjugates, 33 ± 5, 68 ± 21 and 71 ± 10%IA/g, for99mTc-maESE-ZHER2:342, 99mTc-maEES-ZHER2:342 and 99mTc-maSEE-ZHER2:342, respectively, were significantly reduced in comparison with 99mTc-maEEE-ZHER2:342 (102 ± 13%IA/g). For 99mTc-maESE-ZHER2:342, a tumour uptake of 9.6 ± 1.8%IA/g and a tumour-to-blood ratio of 58 ± 6 were reached at 4 h p.i.

Conclusions

A combination of serine and glutamic acid residues in the chelator sequence confers increased renal excretion and relatively low renal uptake of 99mTc-labelled Affibody molecules. In combination with preserved targeting capacity, this improved imaging of targets in abdominal area.

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Acknowledgement

Financial support was provided by the Swedish Cancer Society (Cancerfonden) and the Swedish Research Council (Vetenskapsrådet).

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Authors and Affiliations

  1. School of Biotechnology, Royal Institute of Technology, Stockholm, Sweden

    Torun Ekblad & Amelie Eriksson Karlström

  2. Unit of Biomedical Radiation Sciences, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden

    Thuy Tran, Anna Orlova, Joachim Feldwisch & Vladimir Tolmachev

  3. Affibody AB, Bromma, Sweden

    Anna Orlova, Joachim Feldwisch, Lars Abrahmsén, Anders Wennborg & Vladimir Tolmachev

  4. Section of Hospital Physics, Department of Oncology, Uppsala University Hospital, Uppsala, Sweden

    Charles Widström

  5. Unit of Nuclear Medicine, Department of Medical Sciences, Uppsala University, Uppsala, Sweden

    Vladimir Tolmachev

  6. Biomedical Radiation Sciences, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden

    Vladimir Tolmachev

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  1. Torun Ekblad
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  2. Thuy Tran
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  3. Anna Orlova
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  4. Charles Widström
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  5. Joachim Feldwisch
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  9. Vladimir Tolmachev
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Correspondence to Vladimir Tolmachev.

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Ekblad, T., Tran, T., Orlova, A. et al. Development and preclinical characterisation of 99mTc-labelled Affibody molecules with reduced renal uptake. Eur J Nucl Med Mol Imaging 35, 2245–2255 (2008). https://doi.org/10.1007/s00259-008-0845-7

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  • DOI: https://doi.org/10.1007/s00259-008-0845-7

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