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Metabolic biotinylation of cell surface receptors for in vivo imaging

A Corrigendum to this article was published on 01 August 2006

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We have developed a versatile, potent technique for imaging cells in culture and in vivo by expressing a metabolically biotinylated cell-surface receptor and visualizing it with labeled streptavidin moieties. The recombinant reporter protein, which incorporates a biotin acceptor peptide (BAP) between an N-terminal signal sequence and a transmembrane domain, (BAP-TM) was efficiently biotinylated by endogenous biotin ligase in mammalian cells with the biotin displayed on the cell surface. Tumors expressing the BAP-TM have high sensitivity for magnetic resonance and fluorescence tomographic imaging in vivo after intravascular injection of streptavidin conjugated to magnetic nanoparticles or fluorochromes, respectively. Moreover, streptavidin–horseradish peroxidase conjugates in conjunction with a peroxidase-sensitive gadolinium agent further increased and prolonged the magnetic resonance signal. This BAP-TM allows noninvasive real-time imaging of any cell type transduced to express this reporter protein in culture or in vivo.

*Note: In the version of this article originally published, reference 12 was incorrect. The correct reference 12 is Querol, M., Chen, J.W., Weissleder, R. & Bogdanov, A. Jr. DTPA-bis-amide based MR sensor agents for peroxidase imaging. Org. Lett. 17, 1719–1722 (2005). This error has been corrected in the PDF version of the article.

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Figure 1: Scheme for metabolic biotinylation of surface reporters and imaging of mammalian cells.
Figure 2: Biotinylation of recombinant BAP-TM protein in mammalian cells.
Figure 3: T2-weighted magnetic resonance imaging of tumor cells expressing either biotinylated surface reporter or transferrin receptor reporter in culture and in vivo.
Figure 4: In vivo fluorescence quantitation of tumors expressing metabolically biotinylated mammalian surface receptors.
Figure 5: Magnetic resonance imaging of tumors expressing BAP-TM reporter using a paramagnetic agent that rapidly polymerizes in the presence of peroxidase.

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This work was supported by grants from the US National Cancer Institute CA69246 (X.O.B. and R.W.), CA86355 (R.W. and X.O.B.), CA92782 (R.W.), U01 HL080731 (R.W.), R01 HL078641 (R.W.) and the Brain Tumor Society (X.O.B. and B.A.T.). We thank M. Sena-Esteves (MGH) for providing the lentivirus vectors, P. Waterman for FMT analysis, S. Rhee for magnetic resonance image acquisitions, M. Pittet for help with FACS analysis, N. Sergeyev for synthesizing MNP and F. Reynolds for synthesizing the Gd agent. We also thank S. McDavitt and M. Carlson for editorial assistance.

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Correspondence to Ralph Weissleder.

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Tannous, B., Grimm, J., Perry, K. et al. Metabolic biotinylation of cell surface receptors for in vivo imaging. Nat Methods 3, 391–396 (2006).

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