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PET imaging of CD105/endoglin expression with a 61/64Cu-labeled Fab antibody fragment

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European Journal of Nuclear Medicine and Molecular Imaging Aims and scope Submit manuscript

Abstract

Purpose

The goal of this study was to generate and characterize the Fab fragment of TRC105, a monoclonal antibody that binds with high affinity to human and murine CD105 (i.e., endoglin), and investigate its potential for PET imaging of tumor angiogenesis in a small-animal model after 61/64Cu labeling.

Methods

TRC105-Fab was generated by enzymatic papain digestion. The integrity and CD105 binding affinity of TRC105-Fab was evaluated before NOTA (1,4,7-triazacyclononane-1,4,7-triacetic acid) conjugation and 61/64Cu labeling. Serial PET imaging and biodistribution studies were carried out in the syngeneic 4T1 murine breast cancer model to quantify tumor targeting efficiency and normal organ distribution of 61/64Cu-NOTA-TRC105-Fab. Blocking studies with unlabeled TRC105 were performed to confirm CD105 specificity of the tracer in vivo. Immunofluorescence staining was also conducted to correlate tracer uptake in the tumor and normal tissues with CD105 expression.

Results

TRC105-Fab was produced with high purity through papain digestion of TRC105, as confirmed by SDS-PAGE, HPLC analysis, and mass spectrometry. 61/64Cu labeling of NOTA-TRC105-Fab was achieved with about 50 % yield (specific activity about 44 GBq/μmol). PET imaging revealed rapid uptake of 64Cu-NOTA-TRC105-Fab in the 4T1 tumor (3.6 ± 0.4, 4.2 ± 0.5, 4.9 ± 0.3, 4.4 ± 0.7, and 4.6 ± 0.8 %ID/g at 0.5, 2, 5, 16, and 24 h after injection, respectively; n = 4). Since tumor uptake peaked soon after tracer injection, 61Cu-labeled TRC105-Fab was also able to provide tumor contrast at 3 and 8 h after injection. CD105 specificity of the tracer was confirmed with blocking studies and histological examination.

Conclusion

We report PET imaging of CD105 expression using 61/64Cu-NOTA-TRC105-Fab, which exhibited prominent and target-specific uptake in the 4T1 tumor. The use of a Fab fragment led to much faster tumor uptake (which peaked at a few hours after tracer injection) compared to radiolabeled intact antibody, which may be translated into same-day immunoPET imaging for clinical investigation.

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Acknowledgments

This work was supported in part by the University of Wisconsin Carbone Cancer Center, the Department of Defense (W81XWH-11-1-0644), and the Elsa U. Pardee Foundation.

Conflicts of interest

C.P.T. is an employee of TRACON Pharmaceuticals, Inc. The other authors declare no conflicts of interest.

Author information

Authors and Affiliations

  1. Department of Medical Physics, University of Wisconsin - Madison, 1111 Highland Avenue, Madison, WI, 53705-2275, USA

    Yin Zhang, Hector F. Valdovinos, Todd E. Barnhart & Weibo Cai

  2. Department of Radiology, University of Wisconsin - Madison, Madison, WI, USA

    Hao Hong, Hakan Orbay, Tapas R. Nayak & Weibo Cai

  3. TRACON Pharmaceuticals, Inc., San Diego, CA, USA

    Charles P. Theuer

  4. University of Wisconsin Carbone Cancer Center, Madison, WI, USA

    Weibo Cai

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  1. Yin Zhang
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  2. Hao Hong
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Correspondence to Weibo Cai.

Additional information

Yin Zhang and Hao Hong contributed equally to this work.

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Zhang, Y., Hong, H., Orbay, H. et al. PET imaging of CD105/endoglin expression with a 61/64Cu-labeled Fab antibody fragment. Eur J Nucl Med Mol Imaging 40, 759–767 (2013). https://doi.org/10.1007/s00259-012-2334-2

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  • DOI: https://doi.org/10.1007/s00259-012-2334-2

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