Preparation of specific antisera against adenoviruses by affinity bead immunization (ABI)

Abstract

Certain adenovirus types can be replicated only to low titer in tissue cultures. Other, such as adenovirus strains associated with infantile gastroenteritis, cannot be replicated in vitro. A method which allows preparation of specific antisera has therefore been evaluated. The procedure involves coupling of group-specific antibodies against adenovirus capsid subunits to CNBr-activated Sepharose 4B; reaction of crude virus suspensions with immobilized adenovirus-specific IgG; elimination of contaminating material by extensive washing using a wide pH range; and immunization with adenovirus immunogens immobilized on the beads. Efficient immunization was obtained with immunogen doses of both 50 ng and 50 microgram. The immunization procedure which has been designated affinity bead immunization (ABI) could therefore have a wide applicability in cases where the relevant immunogen constitutes a minor fraction of a crude preparation.