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Signal amplification in molecular imaging by pretargeting a multivalent, bispecific antibody

Nature Medicine volume 11, pages 1250–1255 (2005)Cite this article

Abstract

Here we describe molecular imaging of cancer using signal amplification of a radiotracer in situ by pretargeting a multivalent, bispecific antibody to carcinoembryonic antigen (CEA), which subsequently also captures a radioactive hapten-peptide. Human colon cancer xenografts as small as ∼0.15 g were disclosed in nude mice within 1 h of giving the radiotracer, with tumor/blood ratios increased by ≥40-fold (∼10:1 at 1 h, ∼100:1 at 24 h), compared to a 99mTc-labeled CEA-specific F(ab′) used clinically for colorectal cancer detection, while also increasing tumor uptake tenfold (∼20% injected dose/g) under optimal conditions. This technology could be adapted to other antibodies and imaging modalities.

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Figure 1: Schematic representation of the pretargeting procedure.
Figure 2: Distribution kinetics of the 99mTc-labeled CEA-specific F(ab′) (a) or the 99mTc-radiotracer pretargeted with the divalent CEA-specific × HSG-specific bsMAb (b) or alone (c).
Figure 3: Selecting optimum conditions for pretargeting.
Figure 4: Static imaging using optimal pretargeting conditions.

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Acknowledgements

We thank D. Yeldell, L. Osorio and P.-Y. Brard for their assistance with the animal studies, and N. Velasco for the radiolabeling and quality assurance performed on the reagents used. This work was supported in part by US Public Health Service grant EB002114 from the National Institute of Biomedical Imaging and Bioengineering, US National Institutes of Health, and grant 05-1842-FS-N-0 from the New Jersey Department of Health and Senior Services.

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Authors and Affiliations

  1. Garden State Cancer Center, Center for Molecular Medicine and Immunology, 520 Belleville Avenue, Belleville, 07109, New Jersey, USA

    Robert M Sharkey, Habibe Karacay & David M Goldenberg

  2. Immunomedics, Inc., 300 American Road, Morris Plains, 07950, New Jersey, USA

    Thomas M Cardillo, William J McBride, Hans J Hansen & Ivan D Horak

  3. IBC Pharmaceuticals, Inc., 300 American Road, Morris Plains, 07950, New Jersey, USA

    Edmund A Rossi & Chien-Hsing Chang

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  1. Robert M Sharkey
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Correspondence to David M Goldenberg.

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Competing interests

David M. Goldenberg, Chien-Hsing Chang and Edmund A. Rossi have stock interest or other salary support from IBC Pharmaceuticals, Inc.. David M. Goldenberg, Thomas M. Cardillo, Edmund A. Rossi, Chien-Hsing Chang, William J. McBride, Hans J. Hansen and Ivan D. Horak have stock interest or other salary support from Immunomedics, Inc..

Supplementary information

Supplementary Fig. 1

Quantification of radiotracer uptake during dynamic imaging. (PDF 210 kb)

Supplementary Fig. 2

Emailed author 10/5/05 (PDF 146 kb)

Supplementary Fig. 3

SDS-PAGE analysis of three batches of hBS14 following single-step purification with Affigel-IMP-291 affinity chromatography. (PDF 94 kb)

Supplementary Fig. 4

BIAcore sensorgram showing simultaneous HSG and CEA binding for hBS14 multivalent bsMAb, confirming dual specificity of the bsMAb. (PDF 30 kb)

Supplementary Fig. 5

99mTc-IMP-245 bivalent HSG peptid used as the radiotracer for these studies. (PDF 97 kb)

Supplementary Table 1

The effect of adjusting the bsMAb doe and interval on the uptake of 99mTc radiotracer in the tumor and blood. (PDF 14 kb)

Supplementary Table 2

Biodistribution of 99mTc radiotracer in nude mice bearing GW-39 tumors that were given 500 pmol of the di-bsMAb followed 24 h later with 50 pmol (32 μCi) of the 99mTc radiotracer. (PDF 16 kb)

Supplementary Movie 1

Dynamic 2-min images showing the uptake and elimination of each 99mTc agent over 60 min. (MOV 12376 kb)

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Sharkey, R., Cardillo, T., Rossi, E. et al. Signal amplification in molecular imaging by pretargeting a multivalent, bispecific antibody. Nat Med 11, 1250–1255 (2005). https://doi.org/10.1038/nm1322

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