Summary. High titred monospecific antibodies are obtained in rabbits injecting as little as 25 μg pure antigen per kg body weight. For polyspecific antibodies the dose is increased 10 times. The immunization schedule is described in detail. A procedure is given for the isolation of the immunoglobulins from antiserum by salting out and ion exchange chromatography. Finally antibody titres as measured by 4 different titration methods are compared using anti‐human IgG from rabbits as a model.

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PRINCIPLEThis method is a modified version of the rocket immunoelectrophoresis. Small volumes from fractions obtained in separation experiments are transferred to a row of sample wells in an antibody-free agarose gel, where the applied samples are allowed to diffuse into the gel for 30-60 minutes. The proteins are then by electrophoresis forced into an antibody-containing agarose gel. The antibodies are raised against the antigens that were applied in the preceding separation experiment. The result is that elution profiles for each individual protein are obtained in one single experiment (see Fig. 6.1).

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