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Pharmacokinetics and biodistribution of 177Lu-labeled multivalent single-chain Fv construct of the pancarcinoma monoclonal antibody CC49

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Abstract

Purpose

Lutetium-177 (177Lu) is a radionuclide of interest for radioimmunoimaging (RII) and radioimmunotherapy (RIT) on account of its short half-life (161 h) and the ability to emit both β and γ radiation. Single-chain Fv (scFv) constructs have shown advancement in cancer diagnosis and therapy due to the pharmacokinetics advantage and seem to be intriguing tools in oncology. The objective of this study was to evaluate the pharmacokinetics and biodistribution characteristics of the 177Lu-labeled tetravalent scFv of CC49 MAb and intact CC49 IgG in vivo.

Methods

Conjugation and labeling conditions of multivalent scFv with 177Lu were optimized without affecting integrity and immunoreactivity. For this purpose, multivalent scFv constructs {dimer, sc(Fv)2; tetramer, [sc(Fv)2]2} of the MAb CC49 were expressed as secretory proteins in Pichia pastoris. The purified scFv constructs and IgG form of CC49 were conjugated with a bifunctional chelating agent, ITCB-DTPA, and labeled with 177Lu. The comparative biodistribution, blood clearance, and tumor-targeting characteristics of 177Lu-labeled tetravalent [sc(Fv)2]2 construct of CC49 MAb and intact CC49 IgG were investigated in the athymic mice bearing LS-174T xenografts.

Results

Approximately, 90% of 177Lu incorporation was achieved using ITCB-DTPA chelator, and the labeled immunoconjugates maintained integrity and immunoreactivity. Blood clearance studies demonstrated an alpha half-life (t1/2α) of 177Lu-labeled [sc(Fv)2]2 and IgG of CC49 at 4.40 and 9.50 min and a beta half-life (t1/2β) at 375 and 2,193 min, respectively. At 8 h post administration, the percent of the injected dose accumulated/gram (%ID/g) of the LS-174T tumor was 6.4±1.3 and 8.9±0.6 for 177Lu-labeled [sc(Fv)2]2 and IgG of CC49, respectively, in the absence of l-lysine. The corresponding values were 8.0±0.6 and 8.4±1.2 in the presence of l-lysine. Renal accumulation of [sc(Fv)2]2 was significantly (p<0.005) reduced in the presence of l-lysine.

Conclusion

The results of this study demonstrate that the ITCB-DTPA conjugation and 177Lu-labeling of scFvs are feasible without influencing the antibody characteristics. 177Lu-labeled [sc(Fv)2]2 showed faster clearance and equivalent tumor uptake at 8 h compared with its IgG form, with a markedly reduced renal uptake in the presence of l-lysine. Therefore, 177Lu-labeled [sc(Fv)2]2 may be a potential radiopharmaceutical for the treatment of cancer.

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Acknowledgments

The authors thank Barbara J.M. Booth and Brandon Henley for their technical support. We also thank Dr. Z.P. Kortylewicz, Department of Radiation Oncology, UNMC, for the synthesis and generous supply of ITCB-DTPA, and Ms. Kristi L.W. Berger (Eppley Institute) for editorial assistance. The CC49 scFv construct was a generous gift from Dr. Jeff Schlom of the Laboratory of Tumor Immunology at the National Cancer Institute, NIH, and the Dow Chemical Company. This work was supported by a grant from the United States Department of Energy (DE-FG0295ER62024).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of Nebraska Medical Center, Omaha, NE, 68198, USA

    Subhash C. Chauhan, Maneesh Jain, Erik D. Moore, Uwe A. Wittel & Surinder K. Batra

  2. College of Pharmacy, University of Nebraska Medical Center, Omaha, NE, 68198, USA

    Jing Li & Peter R. Gwilt

  3. Department of Radioimmunotherapy, Beckman Research Institute at City of Hope National Medical Center, Duarte, CA, USA

    David Colcher

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  1. Subhash C. Chauhan
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Correspondence to Surinder K. Batra.

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Chauhan, S.C., Jain, M., Moore, E.D. et al. Pharmacokinetics and biodistribution of 177Lu-labeled multivalent single-chain Fv construct of the pancarcinoma monoclonal antibody CC49. Eur J Nucl Med Mol Imaging 32, 264–273 (2005). https://doi.org/10.1007/s00259-004-1664-0

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  • DOI: https://doi.org/10.1007/s00259-004-1664-0

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