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Fusion of Gaussia Luciferase to an Engineered Anti-carcinoembryonic Antigen (CEA) Antibody for In Vivo Optical Imaging

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Abstract

The bioluminescent protein Gaussia luciferase (GLuc) was fused to an anti-carcinoembryonic antigen (CEA) antibody fragment, the diabody, for in vivo optical tumor imaging. A 15-amino acid N-terminal truncation (GLΔ15) resulted in a brighter protein. Fusions of the anti-CEA diabody to full-length GLuc and GLΔ15 retained high affinity for the antigen, emitted light, and exhibited excellent enzymatic stability. In vivo optical imaging of tumor-bearing mice demonstrated specific targeting of diabody-GLΔ15 to CEA-positive xenografts, with a tumor/background ratio of 3.8 ± 0.4 at four hours after tail-vein injection, compared to antigen-negative tumors at 1.3 ± 0.1 (p = 0.001). MicroPET imaging using 124I-diabody-GLΔ15 demonstrated specific uptake in the CEA-positive tumor (2.6% ID [injected dose]/g) compared to the CEA-negative tumor (0.4% ID/g) at 21 hours. Although further optimization of this fusion protein may be needed to improve in vivo performance, the diabody-GLΔ15 is a promising optical imaging probe for tumor detection in vivo.

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Acknowledgments

The authors wish to thank Dr. Arion Chatziioannou, Dr. David Stout, Waldemar Lladno, and Judy Edwards for assistance with the microPET imaging studies. We appreciate the advice and assistance of Vania Kenanova, Karl Bauer, and Felix Bergara during various stages of the work. We thank Dr. Bruce Bryan for providing plasmids and proteins, and also appreciate the insights of Dr. Bruce Bryan, Dr. Byron Ballou, and Dr. Chris Szent-Gyorgi on the repeated sequence and possible signal peptide in the GLuc gene. This work was supported in part by NIH Grants T32 GM 08652 (K.M.V.), P01 CA 43904 (T.O., A.M.W.), R24 CA92865 (S.S.G.), R01 CA082214 (S.S.G.). A.M.W. is a member of the Jonsson Comprehensive Cancer Center (NIH P30 CA 16042).

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Authors and Affiliations

  1. Crump Institute for Molecular Imaging, Department of Molecular & Medical Pharmacology, David Geffen School of Medicine at UCLA, 700 Westwood Plaza, Los Angeles, California, 90095, USA

    Katy M. Venisnik, Tove Olafsen & Sanjiv S. Gambhir

  2. Molecular Imaging Program at Stanford (MIPS), Department of Radiology and Department of Bioengineering, Stanford University, 300 Pasteur Drive, Stanford, California, 94305, USA

    Anna M. Wu

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  1. Katy M. Venisnik
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  2. Tove Olafsen
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  3. Sanjiv S. Gambhir
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  4. Anna M. Wu
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Correspondence to Anna M. Wu.

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Venisnik, K.M., Olafsen, T., Gambhir, S.S. et al. Fusion of Gaussia Luciferase to an Engineered Anti-carcinoembryonic Antigen (CEA) Antibody for In Vivo Optical Imaging. Mol Imaging Biol 9, 267–277 (2007). https://doi.org/10.1007/s11307-007-0101-8

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  • DOI: https://doi.org/10.1007/s11307-007-0101-8

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