2. Characterization of Endothelial Internalization and Targeting of Antibody—Enzyme Conjugates in Cell Cultures and in Laboratory
Animals
| Abstract |
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Streptavidin-biotin conjugates of enzymes with carrier antibodies provide a versatile means for targeting selected cellular
populations in cell cultures and in vivo. Both specific delivery to cells and proper subcellular addressing of enzyme cargoes
are important parameters of targeting. This chapter describes methodologies for evaluating the binding and internalization
of labeled conjugates directed to endothelial surface adhesion molecules in cell cultures using anti-intercellular adhesion
molecule/catalase or antiplatelet endothelial cell adhesion molecule/catalase conjugates as examples. It also describes protocols
for characterization of biodistribution and pulmonary targeting of radiolabeled conjugates in rats using anti-intercellular
adhesion molecule/tPA conjugates as an example. The experimental procedures, results, and notes provided may help in investigations
of vascular immunotargeting of reporter, experimental, diagnostic, or therapeutic enzymes to endothelial and, perhaps, other
cell types, both in vitro and in vivo.
Affiliation(s): (2) Institute for Environmental Medicine, University of Pennsylvania Medical Center, Philadelphia, PA
Book Title: Bioconjugation Protocols: Strategies and Methods
Series: Methods in Molecular Biology | Volume: 283 | Pub. Date: Jun-10-2004 | Page Range: 21-36 | DOI: 10.1385/1-59259-813-7:021
Subject: Biochemistry
Key Words: Endothelium - cell adhesion molecules - catalase - plasminogen activators - lung targeting
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