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68Ga-labelled recombinant antibody variants for immuno-PET imaging of solid tumours

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Abstract

Purpose

Recombinant antibodies isolated from human antibody libraries have excellent affinities and high target specificity. As full-length IgGs are cleared inadequately slowly from the circulation, the aim of this work was to figure out which kind of recombinant antibody fragment proves to be appropriate for imaging epithelial cell adhesion molecule (EpCAM)-expressing tumours with the short-living radioisotope 68Ga.

Methods

In order to combine the promising tumour targeting properties of antibodies with 68Ga, four antibody variants with the same specificity and origin only differing in molecular weight were constructed for comparison. Therefore, the binding domains of a single-chain fragment variable (scFv) isolated from a human naïve antibody library were modified genetically to construct the respective full-length IgG, the tria- and diabody variants. These molecules were conjugated with the bifunctional chelating agent N,N′-bis[2-hydroxy-5-(carboxyethyl)benzyl]ethylenediamine-N,N′-diacetic acid (HBED-CC) to enable 68Ga labelling at ambient temperature and compared in biodistribution and immuno-PET imaging experiments.

Results

The antibody variants with identical specificity proved to have the correct molecular weight, high binding affinity and specificity to their antigen, EpCAM. Radiometal complexation was efficiently performed at room temperature leading to 68Ga-labelled antibodies with unchanged binding properties compared to the original antibody variants. The best targeting properties were obtained with the scFv and especially with the diabody. The triabody showed higher absolute tumour uptake but only moderate clearance from circulation.

Conclusion

The antibody variants differed considerably in normal organ uptake, clearance from circulation and tumour accumulation. The data demonstrate the feasibility of imaging solid tumours with the 68Ga-labelled diabody format. This type of recombinant protein might be a promising carrier even for the short-lived radiometal 68Ga to support e.g. the management of immunotherapy which may provide important information regarding receptor expression of solid tumours.

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Acknowledgements

A research grant from the Deutsche Krebshilfe for M. Eder is gratefully acknowledged. There is no other financial relationship of the author as stated above. Biodistribution and PET imaging were kindly performed by U. Schierbaum, K. Leotta and U. Bauder-Wüst (all German Cancer Research Center, Heidelberg, Germany). Dr. Moldenhauer (German Cancer Research Center, Heidelberg, Germany) and Prof. Diehl (Cologne, Germany) are gratefully acknowledged for providing cell lines for cell binding analysis. All animal experiments complied with the current laws of the Federal Republic of Germany.

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Authors and Affiliations

  1. Radiopharmaceutical Chemistry, German Cancer Research Center, Im Neuenheimer Feld 280, 69120, Heidelberg, Germany

    Matthias Eder & Michael Eisenhut

  2. Affimed Therapeutics AG, Heidelberg, Germany

    Stefan Knackmuss, Fabrice Le Gall, Uwe Reusch & Melvyn Little

  3. European Molecular Biology Laboratory, Heidelberg, Germany

    Vladimir Rybin

  4. Department of Nuclear Medicine, University of Heidelberg, Heidelberg, Germany

    Uwe Haberkorn & Walter Mier

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  1. Matthias Eder
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  2. Stefan Knackmuss
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Correspondence to Matthias Eder.

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Research grant from the Deutsche Krebshilfe for M. Eder

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Eder, M., Knackmuss, S., Le Gall, F. et al. 68Ga-labelled recombinant antibody variants for immuno-PET imaging of solid tumours. Eur J Nucl Med Mol Imaging 37, 1397–1407 (2010). https://doi.org/10.1007/s00259-010-1392-6

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  • DOI: https://doi.org/10.1007/s00259-010-1392-6

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