Separation of myelin basic protein peptide 43-88 and its fragments by analytic and preparative high-performance...

Reversed-phase (RP) high-performance liquid chromatography (HPLC) using a solvent mixture of triethylammonium formate buffer and methanol permitted the rapid separation of myelin basic protein peptide 43-88 and a mixture of synthetic fragments from this peptide. The elution times for some of the peptides were markedly affected by minimal changes in the solvent mixture. Attempts to separate the same peptides by gel permeation HPLC resulted in poor resolution and an aberrant elution pattern unrelated to molecular size. With the use of the volatile triethylammonium formate buffer in the RP-HPLC, material could be more readily separated and easily recovered by freeze-drying. Analysis of the components separated by this system of RP-HPLC demonstrated that the preparation of normal human BP peptide 43-88 results in an admixture of peptides 43-87 and 43-88. This procedure of RP-HPLC should make it possible to analyze the degradation of myelin basic protein peptide 43-88 and to isolate the degradation products for characterization.