For the first-time generation of antibodies against a particular antigen, we recommend the Multiple-peptide Co-immunization strategy, co-immunize multiple peptide antigens, derived from the same native protein, into the same animal. This two for one approach is intended to enrich the diversity of antibodies that are produced and to short-circuit potential failure if a single antigen does not produce an immune response. While this strategy is not necessarily applicable in all situations, there are real benefits that can be derived from the multiple-peptide immunization strategy.
- Increase the chance for the ability to generate more than one peptide antibodies to one protein due to exposure of greater number of selected target epitopes which can be isolated using specific peptide affinity purification.
- Higher chance of finding best antibodies from the pool of antibodies generated. After isolation, the individual peptide antibodies can be characterized for application such as immunoprecipitation
- Lower cost than separate protocols
This two-for-one approach is intended to enrich the diversity of antibodies that are produced and to short-circuit potential failure if a single antigen does not produce an immune response. In some cases a good polyclonal antibody that can capture the protein from multiple epitopes is produced. However, in practice all antigens are not equal in immunizing potential. Very often the immunodominant peptide will produce the overwhelming majority of antibodies, and little ground is gained for the added expense.
Bio-Synthesis will synthesize, purify and conjugate each peptide separately. Individual KLH conjugates are then mixed in equimolar ratios and used as immunogen. Individual antisera are titered by ELISA against each peptide separately. If client choose to use this strategy, however, one can still use single-anti-peptide strategy by using 2-3 peptides, then conjugate individually and inject to host animal using separate serum packages if budget were not an issue.
See article: Antibody responses to non-immunogenic synthetic peptides induced by co-immunization with immunogenic peptides