Mass-based fraction collection of crude synthetic peptides in analytical and preparative scale

Synthetic peptides become more and more important as drug candidates in the treatment of a variety of diseases. A particular therapeutic focus for synthetic peptides is cancer treatment.1,2 In order to keep pace with the growing number of newly synthesized peptides, peptide purification should not represent the bottleneck in the drug discovery process. Since the target masses of synthetic peptides are well known, mass-based fraction collection represents an efficient technique for their purification. In contrast to fraction triggering with less specific detectors, employing a mass selective detector leads in each run only to the purification of the target mass. Consequently, it is not necessary to pick the compound of interest out of a series of redundant fractions. In this article we demonstrate mass-based purification of a variety of crude synthetic peptides by reversed phase high-performance liquid chromatography. The peptides were in the mass range from less than 1 kDa to more than 10 kDa and covered a pI range from 4 to 13. We particularly focused on some technical aspects of the system that were prerequisite for reliable compound purification with high recoveries.