PEDS Advance Access originally published online on July 31, 2006
Protein Engineering Design and Selection 2006 19(10):453-460; doi:10.1093/protein/gzl030
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Bifunctional antibody-Renilla luciferase fusion protein for in vivo optical detection of tumors
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 2 Molecular Imaging Program at Stanford (MIPS), Department of Radiology and Bio-X Program CA 94305, USA 3 Department of Bioengineering, Stanford University, 300 Pasteur Drive, Stanford CA 94305, USA
4To whom correspondence should be addressed. E-mail: awu{at}mednet.ucla.edu
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
Received January 11, 2006; revised May 3, 2006; accepted June 14, 2006.
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