Tetanus toxoid loaded 'preformed microspheres' of cross-linked dextran

Abstract

The conformation of an antigen and hence its biological activity may get compromised when encapsulated in controlled release microspheres during formulation. In order to obviate the need for exposure of the antigen to the inactivating conditions, such as exposure to organic solvent and the high shear stress of emulsification required for microencapsulation, an alternate strategy was employed. 'Pre-formed' microspheres (20-340 μm in size) made of cross-linked dextran (Dex) were employed as matrix for conjugation of tetanus toxoid (TT) under aqueous conditions. The native immunoreactivity of TT was completely retained after conjugation, as checked by immunofluorescence and quantitative ELISA. Immunogenicity of Dex-TT conjugate was tested in rodents. No untoward mortality or adverse effects of immunization with the test material were observed on histopathology of the site of injection. A single immunization with the long-acting depot formulation elicited anti-TT antibody response lasting for 1 year without any need of booster. The titres were comparable after 12 weeks with those obtained using the conventional alum adsorbed toxoid.