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PEDS Advance Access published online on July 31, 2006
Protein Engineering Design and Selection, doi:10.1093/protein/gzl030
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© The Author 2006. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org
Received January 11, 2006
Revised May 3, 2006
Accepted June 14, 2006

Article

Bifunctional antibody-Renilla luciferase fusion protein for in vivo optical detection of tumors

Katy M. Venisnik 1, Tove Olafsen 1, Andreas M. Loening 2, Meera Iyer 3, Sanjiv S. Gambhir 2, and Anna M. Wu 1 *

1 Crump Institute for Molecular Imaging, Department of Molecular & Medical Pharmacology, David Geffen School of Medicine at UCLA, 700 Westwood Plaza, Los Angeles, CA 90095, USA
2 Molecular Imaging Program at Stanford (MIPS), Department of Radiology and Bio-X Program, Stanford University, 300 Pasteur Drive, Stanford CA 94305, USA; Department of Bioengineering, Stanford University, 300 Pasteur Drive, Stanford CA 94305, USA
3 Crump Institute for Molecular Imaging, Department of Molecular & Medical Pharmacology, David Geffen School of Medicine at UCLA, 700 Westwood Plaza, Los Angeles, CA 90095, USA; Molecular Imaging Program at Stanford (MIPS), Department of Radiology and Bio-X Program, Stanford University, 300 Pasteur Drive, Stanford CA 94305, USA

* To whom correspondence should be addressed.
Anna M. Wu, E-mail: awu{at}mednet.ucla.edu


  Abstract

An anti-carcinoembryonic antigen (CEA) antibody fragment, the anti-CEA diabody, was fused to the bioluminescence enzyme Renilla luciferase (RLuc) to generate a novel optical imaging probe. Native RLuc or one of two stabilized variants (RLucC124A, RLuc8) was used as the bioluminescent moiety. A bioluminescence ELISA showed that diabody-luciferase could simultaneously bind to CEA and emit light. In vivo optical imaging of tumor-bearing mice demonstrated specific targeting of diabody-RLuc8 to CEA-positive xenografts, with a tumor:background ratio of 6.0 ± 0.8 at 6 h after intravenous injection, compared with antigen-negative tumors at 1.0 ± 0.1 (P = 0.05). Targeting and distribution was also evaluated by microPET imaging using 124I-diabody-RLuc8 and confirmed that the optical signal was due to antibody-mediated localization of luciferase. Renilla luciferase, fused to biospecific sequences such as engineered antibodies, can be administered systemically to provide a novel, sensitive method for optical imaging based on expression of cell surface receptors in living organisms.

Keywords: bioluminescence; carcinoembryonic antigen; engineered antibodies; nude mouse xenograft models; Renilla luciferase.
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