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90Y Labeling of monoclonal antibody MOv18 and preclinical validation for radioimmunotherapy of human ovarian carcinomas

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Abstract

The monoclonal antibody (mAb) MOv18 binds the membrane alpha isoform of the folate receptor (FR) which is overexpressed in human ovarian carcinoma cells. Exploiting the targeting capacity of this mAb, we developed and preclinically validated a protocol for the stable labeling of the mAb with 90Y, an isotope which has shown promise in cancer radioimmunotherapy. MOv18 was derivatized with the stable macrocyclic ligand p-isothiocyanatobenzyl-1,4,7,10-tetraazacyclododecane-1,4,7,10- tetraacetic acid (Bz-DOTA). MOv18-Bz-DOTA conjugates were labeled with 90Y or 111In under metal-free and good laboratory practice conditions. At the optimal Bz-DOTA/mAb derivatization ratio of 4–5, conjugates maintained binding activity up to 6 months, were efficiently labeled with 90Y or 111In (mean labeling yield 85 and 64%, associated to a final mean specific activity of 74 and 37 MBq/mg) and displayed a mean immunoreactivity of 60 and 58%, respectively. The radiolabeled preparations were stable in human serum, with >97% radioactivity associated to mAb at 48 h after labeling. The ability of 90Y- and 111In-MOv18 to localize FR on tumors in vivo was analyzed in nude mice bearing tumors induced by isogenic cell lines differing only in the presence or absence of the relevant antigen [A431FR (FR-positive) and A431tMock (FR-negative)]. In vivo biodistribution in organs other than tumor was comparable in non-tumor-, A431tMock- and A431FR-bearing mice, whereas the median tumor uptake of the radiolabeled reagents, expressed as area under the curve (AUC) and maximum uptake (Umax), was significantly higher (sixfold to sevenfold) in A431FR than in A431tMock tumors (P=0.0465 and P=0.0332, respectively). Mean maximum uptake (% ID/g) for 90Y-MOv18 was 53.7 and 7.4 in A431FR and A431tMock respectively; corresponding values for 111In-Mov18 were 45.0 and 11.3. These data demonstrate the feasibility of 90Y-labeling of MOv18 without compromising antibody binding ability and the immunoreagent-specific localization in vivo on FR-expressing tumors, suggesting the suitability of 90Y-MOv18 for clinical studies.

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Abbreviations

mAb:

Monoclonal antibody

FR:

Alpha isoform of the folate receptor

Bz-DOTA:

p-isothiocyanatobenzyl-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid

GLP:

Good laboratory practice

% ID/g:

Percent injected dose/g

AUC:

Area under the curve

Umax:

Maximum uptake

HSA:

Human serum albumin

SDS-PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

MALDI-TOF-MS:

Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

L/P:

Ligand/protein ratio

FITC:

Fluorescein-isothiocyanate

DTPA:

Diethylenetriaminepentaacetic acid

EDTA:

Ethylenediaminetetraacetic acid

ITLC:

Instant thin layer chromatography

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Acknowledgements

The authors thank Gloria Bosco for manuscript preparation. This work was partially supported by AIRC-FIRC and a MIUR grant to Dompé.

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Authors and Affiliations

  1. Unit of Nuclear Medicine, Istituto Nazionale Tumori, via Venezian 1, 20133, Milano, Italy

    Angela Coliva, Ettore Seregni & Emilio Bombardieri

  2. Unit of Molecular Therapies, Department of Experimental Oncology, Istituto Nazionale Tumori, via Venezian 1, 20133, Milano, Italy

    Alberto Zacchetti, Elena Luison, Antonella Tomassetti, Mariangela Figini & Silvana Canevari

  3. Molecular Mechanisms of Cancer Growth and Progression, Istituto Nazionale Tumori, via Venezian 1, 20133, Milano, Italy

    Italia Bongarzone

  4. Dompe’ S.p.a., L’Aquila, Italy

    Franck Martin

  5. Physics Department, Università degli Studi di Milano, Milano, Italy

    Augusto Giussani

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  1. Angela Coliva
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  2. Alberto Zacchetti
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  3. Elena Luison
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  9. Augusto Giussani
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  10. Mariangela Figini
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  11. Silvana Canevari
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Correspondence to Silvana Canevari.

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Angela Coliva and Alberto Zacchetti contributed equally to this work.

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Coliva, A., Zacchetti, A., Luison, E. et al. 90Y Labeling of monoclonal antibody MOv18 and preclinical validation for radioimmunotherapy of human ovarian carcinomas. Cancer Immunol Immunother 54, 1200–1213 (2005). https://doi.org/10.1007/s00262-005-0693-2

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  • DOI: https://doi.org/10.1007/s00262-005-0693-2

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