There is much to be said on the topic of antibody development and peptide design considerations. Here however we will focus on the production of peptides where these are used as antigens. One could play it safe and go for high purity peptides for every project, but depending on the target and on the synthesis strategy, this is seldom necessary and thus there is time and money to be saved.
Is the target depending on a specific site? For example, a post translational modification such as phosphorylation, a neo-epitope, or an exon bridge defining a specific isoform. In such a case high purity is often a good idea – but it may not always be necessary.
A key point is knowing how the peptide is synthesised. If a capping cycle is run between each amino acid coupling step, then much is gained. However, since this adds to the production cost, few vendors will do this by default (you’ll have to explicitly ask for it). The advantage is that, regardless of purity, the impurities largely consist of N-terminally truncated sequences. That means that 100% of the material is of the correct sequence, albeit some shorter than full length. But since this is a polyclonal antibody, it is not much of an issue. On the other hand, without the capping you will have deletion sequences, and you will effectively immunise with X% of peptide with the wrong sequence.
Since peptides are synthesized from C-terminus to N-terminus, the strategy of employing a capping cycle for each amino acid coupling step means that when the peptide is conjugated to the carrier molecule via its N-terminus, only full length peptide will take part in the reaction - regardless of purity.
In the cases where high purity peptide is called for, e.g. for monoclonal antibody development, you have to think quality in all steps of the production; raw materials, synthesis strategy, and quality control. We’ll invite you to read about our view on the production of high quality peptides.
Innovagen provides a full range of custom services for both antigen and antibody production.
Innovagen has serviced the research community since 1992. In that time we have delivered a numerous antibodies raised against synthetic peptides. Our experience and commitment to quality and customer service are all things that you will benefit from when turning to us for your peptide requirements.
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