Choosing the Solid Phase Extraction (SPE) Media for Synthetic Peptide Clean Up

Parallel peptide app note - sorbent selection when doing 96 well plate peptide library purification.

Solid phase extraction (SPE) is a commonly used method to clean up various types of samples before analyzing them using LCMS. It is also an effective way to improve the purity of crude synthetic peptide samples, which significantly reduces the time required to deliver compounds for evaluation. Additionally, this method reduces solvent consumption by over 90% compared to standard sequential HPLC purification, which achieves purities typically not required for secondary screening assays. Importantly, peptide clean up methodologies utilizing SPE in a 96-well format can be automated, allowing large numbers of samples to be processed in parallel and complementing high throughput peptide synthesis methodologies commonly employed. Taken together, these features make SPE an excellent technique for purifying peptide libraries.

However, to achieve these benefits, it is essential to select a media in the well plate that interacts predictably with the peptide samples – eluting in high yield with a single mobile phase treatment, thereby eliminating the need to analyze every sample prior to the cleanup process and every fraction generated therein. Biotage has developed the PeptiRen-96 C18 well plate for peptide library cleanup. In this study, we show that using Biotage®  PeptiRen C18 media results in significantly improved purity of crude samples in a predictable manner compared to other commercially available media.