Conjugation of catalase to a carrier antibody via a streptavidin-biotin cross-linker.
Targeting of catalase could be useful in antioxidative protection of cells challenged with H2O2. In the present study we conjugated catalase to a carrier model antibody using a biotin-streptavidin (SA) cross-linker and characterized the functional activity of the conjugate. Neither biotinylation nor conjugation with SA decreased the enzymic activity of catalase. Further coupling of radiolabelled biotinylated catalase (b-catalase) to biotinylated antibody (b-Ab) via SA using a two-step conjugation procedure did not change enzymic activity of b-catalase. b-Ab-SA-b-catalase specifically bound to antigen-coated plastic wells, but not to albumin-coated plastic wells. Substitution of b-Ab with control biotinylated IgG (b-IgG) abrogated binding of the catalase to the antigen. H2O2 was degraded in antigen-coated wells preincubated with b-Ab-SA-b-catalase, but not with b-IgG-SA-b-catalase. Thus b-Ab-SA-b-catalase specifically binds to immobilized antigen and degrades H2O2 after binding to the target. The methodology described in the present paper may be useful for the development of a novel strategy for antioxidant therapy.
Muzykantov
V R
VR
Institute for Environmental Medicine, University of Pennsylvania Medical School, Philadelphia, USA.
D000818:Animals; D000906:Antibodies; D000975:Antioxidants / Q000008:administration & dosage / Q000378:metabolism; D001709:Biotechnology; D001710:Biotin; D002374:Catalase / Q000008:administration & dosage* / Q000378:metabolism; D002417:Cattle; D003432:Cross-Linking Reagents; D006041:Goats; D006861:Hydrogen Peroxide / Q000378:metabolism; D018796:Immunoconjugates / Q000008:administration & dosage* / Q000302:isolation & purification; D066298:In Vitro Techniques; D051379:Mice; D019809:Streptavidin
D000906:Antibodies; D000975:Antioxidants; D003432:Cross-Linking Reagents; D018796:Immunoconjugates; D001710:Biotin; D019809:Streptavidin; D006861:Hydrogen Peroxide; D002374:Catalase