Making antigens immunogenic

There is no perfect recipe for the generation of a good antiserum. Many foreign molecules (viruses, bacteria and cells) elicit strong antibody responses but some foreign substances fail to induce a good antiserum. In some of these instances the immune system can be manipulated to generate a response, and this can be achieved either by modifying the antigen or varying the host animal. Many of these modifications have been established empirically over the years; however, our increased understanding of the molecular events controlling an immune response should allow more rational approaches for the generation of high titer, specific antisera. Before attempting to produce an antiserum, consideration should be given to the following points: the nature of the antigen and its purity, the site/route of immunization, the dose of the antigen, the choice of adjuvants and the genetic background of the animal to be immunized.

In general, particulate antigens and higher molecular weight proteins tend to be more immunogenic than soluble antigens and proteins of low molecular weight. Converting small soluble molecules into large aggregates is an effective way of enhancing their immunogenicity. The physical coupling of a small molecule which is nonimmunogenic on its own (hapten) to a larger immunogenic carrier molecule also achieves the same result. The most common routes of immunization are subcutaneous, intramuscular, intradermal, intraperitoneal and intravenous. Other routes of immunization include oral ingestion of antigen, antigen inhalation, topical application to the skin and the direct injection into an organ such as the spleen. The dose of the antigen to be administered can profoundly influence the outcome of the procedure. Thus either very low or high doses of antigen may induce tolerance rather than the successful production of an antiserum. One should also bear in mind that with multiple injections, trace contaminants within an antigenic preparation may eventually elicit responses which will be detected in later bleeds collected from the animal and may result in an antiserum with multiple specificities. The degree of specificity of an antiserum depends largely on the degree of purity of the antigen used. The decision about antigen purity is often based on the intended use of the antiserum. If a highly specific antiserum that will only recognize the appropriate antigen is needed, then before commencing immunization the antigen must be purified to homogeneity.

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