Making Weak Antigens Strong: Modifying Protein Antigens by Denaturation

Many molecules can be made more immunogenic by denaturation. This treatment will change the structure of many compounds, particularly proteins, and expose new epitopes. In addition, heating will often cause protein antigens to aggregate, and, because aggregated antigens are often more immunogenic, this can increase the antibody response. Either heating alone or heating with sodium dodecyl sulfate (SDS) is the usual treatment. Injecting denatured antigens will be more likely to produce an antibody response against epitopes that are not found on the native antigen. If antibodies against totally denatured proteins are desired, the SDS-heat treatment is normally best. These treatments will generate antibodies that should be particularly good for immunoblots, for screening bacterial expression libraries, and for immunoprecipitation of proteins synthesized during in vitro translations.