The generation of custom antibodies can be accomplished by using one of two primary approaches. The first is the use of full length protein as immunogen. This could consist of either native, recombinant, or fusion protein, or even protein extracted from a gel band. The second option is to generate short peptides from specifically chosen, short sequences of the native protein and use these as immunogen instead of a full length protein. In recent years, the use of such synthetic peptide antigens as a means to generate custom antibodies has markedly increased. Here we outline a number of advantages to the use of peptide antigens, as well as several factors that affect successful anti-peptide antibody production.
In cases where access to a target protein is limited or unavailable, but the amino acid sequence is known, peptide antigens offer a smart solution for antibody generation. Peptide antigens are also ideal for producing antibodies against post-translational modification sites within a target protein. In fact, Phospho-Specific Antibodies developed by our expert scientists detect specific protein phosphorylation with high sensitivity even within complex cellular protein mixtures.
Additional Advantages of Peptide Antigens
Lower probability of cross-reactivity
More flexibility in antigen selection
Only protein sequence is needed
Easy to synthesize and cost-effective
Fast turnaround time
Epitope well-defined from beginning
As you begin your custom antibody project, it is important to consider that while many peptide sequences can be immunogenic, not all are equally effective at generating antibodies with reactivity against the target protein. Factors affecting successful antibody production using peptide antigens include: the accuracy of the target protein sequence, the predicted secondary structure of the intact protein, the coupling approach of the peptide antigen to a larger carrier protein, and the ease of synthesis of specific peptide sequences. Click here to learn more about peptide antigen design.
GenScript offers several unique strategies to increase the chances of producing anti-peptide antibodies which recognize native protein. The first is our proprietary OptimumAntigen design tool that combines top-notch algorithms with proven expertise to help design the optimal peptide antigen for your project. Each peptide antigen is measured against several databases to confirm the desired antibody and epitope specificity, ultimately finding the best candidate peptides with antigenicity and solubility suitable for antibody production. This state-of-the-art OptimumAntigen design tool also allows us to offer guaranteed ELISA titers of at least 1:32,000 for any antibody package.
The second strategy is the use of carrier proteins for peptide immunization. A peptide antigen with a low molecular weight is not sufficient to stimulate antibody generation from the immune system of a host animal without the conjugation of a carrier protein such as KLH, BSA or OVA. GenScript's experienced scientists provide intelligent and diverse approaches to peptide conjugation for effective antibody production.
The final strategy is the use of multiple peptide antigens from a single target protein. Multiple peptide antigens, designed by OptimumAntigen, have been proven to be an effective way to markedly increase the success rate of native protein detection. In our experience, antibodies generated using three different peptide antigens against one protein can achieve a 95% success rate of Western blot endogenous protein detection.
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