Abstract
Purpose
The new antibody pertuzumab (Omnitarg) targets the dimerisation subdomain of HER-2. The purpose of this study was to analyse whether pertuzumab retains HER-2 targeting capacity after labelling with the therapeutically interesting beta emitter 177Lu and to make initial characterisations in vitro and in vivo.
Methods
Pertuzumab was conjugated with isothiocyanate-benzyl-CHX-A″-DTPA and chelated to 177Lu. Immunoreactivity, affinity, cellular retention and internalisation were analysed using SKOV-3 cells. The affinity of non-radioactive pertuzumab was measured using a surface plasmon resonance biosensor. In vivo targeting and specific binding were assessed in Balb/c (nu/nu) mice carrying SKOV-3 xenografts. The biodistribution of 177Lu was determined 1, 3 and 7 days after [177Lu]pertuzumab administration. Gamma camera images were taken after 3 days.
Results
The immunoreactivity of [177Lu]pertuzumab was 85.8±1.3%. The affinity of non-radioactive pertuzumab was 1.8±1.1 nM, and that of [177Lu]pertuzumab, 4.1±0.7 nM. The cellular retention after 5 h pre-incubation was 90±2% at 20 h. The targeting was HER-2 specific both in vitro and in vivo, since excess amounts of non-labelled antibody inhibited the uptake of labelled antibody (p<0.0001 and p<0.01, respectively). The biodistribution and gamma camera images of 177Lu showed extensive tumour uptake. Normal tissues had a surprisingly low uptake.
Conclusion
Pertuzumab was efficiently labelled with 177Lu and showed good intracellular retention and HER-2 specific binding both in vitro and in vivo. The gamma camera images and the biodistribution study gave excellent tumour targeting results. Thus, [177Lu]pertuzumab is of interest for further studies aimed at radionuclide therapy.
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Acknowledgements
The authors thank Professor Hans Lundqvist for advice on nuclide techniques and Veronica Asplund Eriksson for technical support. Thanks are also expressed to Genentech for the generous gift of Omnitarg. This work was financially supported by the Swedish Cancer Society.
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Persson, M., Tolmachev, V., Andersson, K. et al. [177Lu]pertuzumab: experimental studies on targeting of HER-2 positive tumour cells. Eur J Nucl Med Mol Imaging 32, 1457–1462 (2005). https://doi.org/10.1007/s00259-005-1902-0
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DOI: https://doi.org/10.1007/s00259-005-1902-0