Streptavidin-Biotin Crosslinking of Therapeutic Enzymes With Carrier Antibodies: Nanoconjugates for Protection Against Endothelial Oxidative Stress

The streptavidin-biotin system may be used to synthesize immunoconjugates for targeted delivery of drugs, including therapeutic enzymes. The size of antibody-enzyme conjugates, which is controlled by the extent of biotinylation and molar ratio between the conjugate components, represents an important parameter that in some cases dictates subcellular addressing of drugs. This chapter describes the methodology of formation and characterization of polymeric immunoconjugates in the nanoscale range. A theoretical model of streptavidin conjugation based on general principles of polymer chemistry is considered. Factors that influence size and functional characterization of resulting polymer conjugates, as well as advantages and limitations of this approach, are described in detail. The protocols describe the formation of immunoconjugates possessing an antioxidant enzyme, catalase, directed to endothelial cells by anti-platelet endothelial cell adhesion molecule antibodies. However because of the modular nature of the streptavidin-biotin crosslinker system, the techniques herein can be easily adapted for the preparation of nanoscale immunoconjugates delivering other protein drugs to diverse cellular antigens.