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

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.

Identification of an enteric adenovirus by immunoelectroosmophoresis (IEOP) technique.

Infantile gastroenteritis can be caused by several of the established adenovirus types. There are also adenoviruses that defy in vitro cultivation but have been recognized by electron microscopy. One isolate of these viruses, which have been designated enteric adenoviruses, has been characterized. Several of the established adenovirus types are shed in stools over long periods. The conditions for direct identification of enteric adenoviruses by immunoelectroosmophoresis (IEOP) have therefore been evaluated. Application of this technique requires highly specific reagents. However, a high prevalence of antibodies in rabbits reacting with human rotavirus was noted. For these reasons, immunoadsorbent purified antibodies were prepared. Because of the difficulty in purifying immunogens from stools, an immunization procedure characterized by immunization with adenovirus subunits bound to affinity chromatography beads was elaborated. An identification procedure for adenoviruses causing infantile gastroenteritis based a) on IEOP using group-specific and monospecific antibodies and b) on the determination of the in vitro cultivatability is suggested.