Macro-Prep HIC Support Datasheet

Hydrophobic interaction chromatography (HIC) separates proteins on the basis of relative hydrophobicity. HIC is a natural second step after either ion exchange or salt precipitation since the sample is applied in high-salt buffer. At high ionic strengths, hydrophobic sites of the protein interact with the alkyl groups of the support. Retention, selectivity, and biological activity are somewhat dependent on pH, type of salt used, and its concentration.

The Macro-Prep methyl HIC support is ideal for purification of proteins with strongly hydrophobic regions. The Macro-Prep t-butyl HIC support is ideal for purification of proteins with few or weakly hydrophobic regions. The two different ligands provide alternative selectivities for easier optimization of separation (Figure 1 and Table 1). The properties of the supports are summarized in Table 2. These methacrylate copolymer beads provide high resolution at very high flow rates. They can be sanitized quickly and efficiently in 0.15% peracetic acid (Figure 2), and are compatible with many common solutions (Table 3). Changes in pH or ionic strength of the buffer do not cause shrinking or swelling of the support.