Use of direct fluorescence labeling and confocal microscopy to determine the biodistribution of two protein therapeutics,Cerezyme® and Ceredase® |
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Authors: | Peter A. Piepenhagen Scott Vanpatten Heather Hughes James Waire James Murray Laura Andrews Tim Edmunds Michael O'Callaghan Beth L. Thurberg |
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Affiliation: | 1. Department of Pathology, Genzyme Corporation, Framingham, Massachusetts 01701;2. Department of Structural Protein Chemistry, Genzyme Corporation, Framingham, Massachusetts 01701;3. Department of Immunology, Genzyme Corporation, Framingham, Massachusetts 01701;4. Department of Pharmacology and Toxicology, Genzyme Corporation, Framingham, Massachusetts 01701;5. Preclinical Biology Genzyme Corporation, Framingham, Massachusetts 01701 |
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Abstract: | Efficient targeting of therapeutic reagents to tissues and cell types of interest is critical to achieving therapeutic efficacy and avoiding unwanted side effects due to offtarget uptake. To increase assay efficiency and reduce the number of animals used per experiment during preclinical development, we used a combination of direct fluorescence labeling and confocal microscopy to simultaneously examine the biodistribution of two therapeutic proteins, Cerezyme® and Ceredase®, in the same animals. We show that the fluorescent tags do not interfere with protein uptake and localization. We are able to detect Cerezyme and Ceredase in intact cells and organs and demonstrate colocalization within target cells using confocal microscopy. In addition, the relative amount of protein internalized by different cell types can be quantified using cell type‐specific markers and morphometric analysis. This approach provides an easy and straightforward means of assessing the tissue and cell type‐specific biodistribution of multiple protein therapeutics in target organs using a minimal number of animals. Microsc. Res. Tech. 2010. © 2009 Wiley‐Liss, Inc. |
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Keywords: | confocal microscopy morphometry protein biodistribution |
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