ELISA-Entwicklung GmbH & Co. KG
The laboratory for professional assay development
Assay Development using Synthetic Peptides
Antibody production against a synthetic peptide is useful if the natural antigen is not available or if it is unstable or in support of the purification programme. Further applications of this approach may include production of antibodies with high specificity for a particular amino acid sequence; this is a useful approach for protein mapping studies or for assays with high specificity. For this purpose a partial sequence of approximately 15-20 amino acids will be selected from protein data bases.Antisera against a synthetic peptide differ from antisera against the complete protein in the following manner:
- High specificity often comparable to monoclonal antibodies.
- Affinity is sometimes lower in comparison with polyclonal antisera against the complete protein.
- Usefulness of the synthetic peptide as standard or tracer in a quantitative test may be limited, due to differences in cross-reactivity or affinity of the synthetic peptide in comparison with the native protein.
- Longer immunisation times for production of synthetic peptide antisera in rabbits may be encountered.
Peptide synthesis and antibody production are outsourced to another specialised laboratory.
Despite the described disadvantages peptide antibodies are a very good and highly specific biochemical tool during protein purification. Sometimes, they are the only realistic possibility for test development of a given protein.
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