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Peptide Synthesis >
Purification
Purification of Synthetic Peptides
Crude peptides are purified by reverse phase
HPLC on a YMC, C-18 column (ODS-A, 2.0 cm x 25 cm, 120 angstrom pore size, 5
micron particle size) that is equilibrated in 0.05% TFA at a flow rate of 7.0
ml/min. The standard linear gradient increases from 0% to 100% acetonitrile over
300 min. Generally, however, only a portion of this gradient (centered around
the expected acetonitrile concentration needed for elution) is run. To ensure
that the highest purity fractions are pooled, aliquots of peak fractions are
subjected to matrix assisted laser desorption ionization mass spectrometry (MALDI-MS)
and analytical HPLC, which provides higher resolution than can be attained on
our preparative HPLC. After pooling the highest purity fractions together, they
are lyophilized to dryness.
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