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Near Infrared Technology and Optical Agents For Molecular Imaging

1, 8.



In a cell based assay system using IRDye 800CW labeled secondary antibodies to assess cell signaling pathways, IRDye 800CW was shown to yield superior signal-to-background ratios and enabled quantification of low levels of protein phosphorylation9.

In contrast, the signal to background using Cy5.5- labeled secondary antibodies was too low to allow its use in the assay (data not shown). Further experiments showed that Cy5.5 exhibited a high level of non-specific binding to cells (data not shown). This observation, coupled to the fact that Cy5.5 is outside the optimal NIR region, makes it less suitable for imaging studies requiring high signal to background.

The performance of IRDye 800CW has been compared to radiochemical detection in animal studies. Using gamma scintigraphy and NIR imaging, Houston et al . compared the ability of a cyclopentapeptide dual-labeled with 111indium and I R D ye 800CW to image av3-integrin positive melanoma xenografts10. The tumor regions were clearly delineated by optical imaging of the IRDye 800CW signal. In contrast, the tumor boundaries could not be identified by scintigraphy due to high noise levels.

We have successfully used IRDye 800CW conjugated to epidermal growth factor (IRDye 800CW-EGF) as an optical agent for imaging tumor progression11. Figure 2 shows an example of a prostate tumor-bearing SCID mouse injected with IRDye 800CWlabeled EGF. The IRDye 800CW-EGF showed good sensitivity with very low background from autofluorescence or non-specific binding.



For animal imaging, targeted NIR-labeled ligands can be used to visualize virtually any pathology without the need for engineered cell lines as required for imaging by bioluminescence or f
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