Peptide synthesis for preparing antisera
How long should the peptides be?
Peptides from 10 to up to 20 amino acids are useful for antibody development.
How precise is the prediction of the peptide sequence which will be suitable as an epitope?
The prediction is based on the amino acid sequence of the protein and uses a computer program which employs various mathematical models.
These models can only simulate the natural structure of the protein without taking modifications (e.g. phosphorylations, glycosylation, S-S bonds) into consideration. For this reason, peptide sequences which are found being suitable as epitopes might not work against the respective protein in the customers test system(s), even when antibodies can be generated against them.
In general there are several possible sequences you can choose from. In case of doubt, it is advisable to select more than one peptide for antibody development.
When making the prediction, the following factors are investigated:
- hydrophobicity
- hydrophilicity
- helical membranous regions
- solvent accessibility
- antigenicity
Why do peptides have to be coupled to a carrier protein?
The peptide itself is too small to be recognized by the animal, therefore it has to be coupled to a protein carrier in order to gain an immune response.
How are the peptides coupled?
There are several possibilities:
- undirected (random) coupling by way of the NH2 or COOH groups
- directed coupling by way of the cysteine SH groups
We recommend directed coupling by way of a cysteine in order to obtain a specific confirmation for the peptide. If the peptide sequence does not contain a natural N-terminal or C-terminal cysteine, a cysteine should be added on during the synthesis.
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