Methodology for purification of large hydrophobic peptides by high-performance liquid chromatography.

To aid in structural studies of pig cardiac myosin light chains (L27 and L28), a procedure of ion-exchange chromatography (IEC) on Trisacryl M (noted for its high capacity) in combination with reversed-phase high-performance liquid chromatography (RP-HPLC) and volatile buffers has been developed. In contrast with other IEC methods (resins or HPIEC), the use of Trisacryl M facilitates subsequent peptide purifications by RP-HPLC. The advantage of the present combination of techniques is also that it enables the isolation of hydrophobic peptides in high yield, e.g., the N-terminal chymotryptic peptide from L27 was thus purified and, after sub-digestion with trypsin, its sequence has been established.