2] Production of antisera with small doses of immunogen: multiple intradermal injections

Abstract

This chapter describes the production of antisera with small doses of immunogen. A wide variety of immunization techniques has been used to generate specific antisera in laboratory animals. Those techniques incorporate a variety of injection routes, vehicles, and frequencies of injection into appropriate laboratory animals. The immunogen is initially dissolved in a buffer at an appropriate pH and molarity to enhance solubilization of the immunogen in that aqueous solution. An equal volume of that solution is combined with Freund's complete or incomplete adjuvant. It is suggested that, when using Freund's complete adjuvant, which contains per milliliter 2 mg of heat-killed tubercle bacillus, one must be certain that the insoluble mycobacterium is suspended in the oil. One of the primary factors, governing the selection of the animal species, is the antigenic similarity between the substance to be injected and that present in the animal species to be injected. As a generalization, antibody titers are readily maintained or increased with booster injections, but the specificity of the antisera may be lost or markedly changed with repeated injections.