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Conjugation and radiolabeling of monoclonal antibodies with zirconium-89 for PET imaging using the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine

Nature Protocols volume 5, pages 739–743 (2010)Cite this article

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Abstract

The positron emitter zirconium-89 (89Zr) has very attractive properties for positron emission tomography (PET) imaging of intact monoclonal antibodies (mAbs) using immuno-PET. This protocol describes the step-by-step procedure for the facile radiolabeling of mAbs or other proteins with 89Zr using p-isothiocyanatobenzyl-desferrioxamine (Df–Bz–NCS). First, Df–Bz–NCS is coupled to the lysine–NH2 groups of a mAb at pH 9.0 (pre-modification), followed by purification using gel filtration. Next, the pre-modified mAb is labeled at room temperature by the addition of [89Zr]Zr-oxalic acid solution followed by purification using gel filtration. The entire process of pre-modification, radiolabeling and purification steps will take about 2.5 h.

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Figure 1
Figure 2: Representative HPLC chromatograms and SDS-PAGE gels.
Figure 3: Positron emission tomography (PET) images of a nude mouse bearing two head and neck cancer FaDu xenografts, obtained at 72 h after i.v. injection of the anti-CD44v6 conjugate 89Zr–Df–Bz–NCS–cmAb U36 (3.7 MBq, 200 μg mAb) with a double-crystal-layer high-resolution research tomography PET scanner (HRRT PET scanner, Siemens/CTI).

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Acknowledgements

This project was financially supported by the Dutch Technology Foundation (STW, grants VBC.6120 and 10074) and partly supported by the European Union FP7, ADAMANT, and partly carried out within the framework of CTMM, the Center for Translational Molecular Medicine, project AIRFORCE number 030-103. The authors thank the technical staff of BV Cyclotron VU and the Radionuclide Centre for supply and processing of 89Zr.

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Authors and Affiliations

  1. Departments of Otolaryngology/Head and Neck Surgery, VU University Medical Center, Amsterdam, The Netherlands

    Maria J W D Vosjan, Lars R Perk, Marianne Budde & Guus A M S van Dongen

  2. Nuclear Medicine & PET Research, VU University Medical Center, Amsterdam, The Netherlands

    Gerard W M Visser & Guus A M S van Dongen

  3. Macrocyclics, Dallas, Texas, USA

    Paul Jurek & Garry E Kiefer

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  1. Maria J W D Vosjan
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Contributions

M.J.W.D.V. and L.R.P. designed and carried out the in vitro and in vivo experiments and prepared the manuscript. M.B. carried out the in vitro experiments. G.W.M.V. and G.A.M.S.v.D. designed the overall approach, advised in chemical aspects of the experiments and edited the manuscript. P.J. and G.E.K. designed and synthesized the chelate and advised in the experiments.

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Correspondence to Guus A M S van Dongen.

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Competing interests

P.J. and G.E.K. are employees of Macrocyclics, the vendor of the chelate.

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Vosjan, M., Perk, L., Visser, G. et al. Conjugation and radiolabeling of monoclonal antibodies with zirconium-89 for PET imaging using the bifunctional chelate p-isothiocyanatobenzyl-desferrioxamine. Nat Protoc 5, 739–743 (2010). https://doi.org/10.1038/nprot.2010.13

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