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Immunization, Isolation of Immunoglobulins and Antibody Titre Determination

During the past 10 years we have studied the immunization of several thousand outbred rabbits with a multitude of immunogens and analysed the resulting antisera. It is obvious that an immunization procedure that is optimal in all situations and guarantees a good antibody response in every animal does not exist. The aim of this chapter is to mention some of the factors that influence the antibody response and to describe an immunization scheme that has a statistically fair chance of giving good antisera. We also describe a simple procedure for the isolation of the immunoglobulin fraction of rabbit antiserum and compare some methods for antibody titre determination and determination of antibody specificity. Throughout the chapter the emphasis is on precipitating antisera to macromolecular antigens.