PRP-3 polymeric reversed phase columns last longer than silica-based columns. The columns are packed with a poly(styrene-divinyl)benzene support. The support is pressure stable up to 5,000 psi and crosslinked to prevent shrinkage or swelling (common in other polymeric HPLC columns) when the mobile phase is changed. Since the support is entirely polymeric and there are no C18 groups to hydrolyze, column lifetime is longer. Regardless of whether you use ionpair reagents or high percentage aqueous buffers (80-100%) for your separation, PRP-3 columns outlast silica-based columns under common (pH 2.0) or uncommon (pH 12.7) conditions. There can be sample artifacts in protein samples that will not elute under regular separation conditions. After continued use these artifacts will accumulate and cause an increase in back pressure and a decrease in column performance. When this occurs a simple overnight regeneration process washes away the sample artifacts and restores your column to peak performance.

Superior pH Stability
While most reversed phase columns are slowly destroyed by buffers below pH 2.0 or above 8.0, the inherent stability of PRP-3 columns allows their use for any protein purification method from pH 1 to 13. This is very helpful for proteins that are unstable or have poor solubility at a pH below 8.0. While most proteins are best chromatographed at pH 2.0, it's good to know you can separate them at elevated pH if the need arises. PRP-3 columns allow you to analyze protein purity at low and high pH extremes. Applications #7 and #250 illustrate the separation of a protein mixture at pH 2.0 and 12.7. At pH 2.0 the order of elution is Bradykinin, Ribonuclease A and Insulin. When the same sample is chromatographed at pH 12.7, the elution order has changed to Ribonuclease A, Insulin, and Bradykinin.

Protein Recovery
The 300 Angstrom pores on the PRP-3 support help ensure good recovery (>90%) of your protein. The highly inert poly(styrene-divinylbenzene) packing enhances protein recovery because there are no silanol groups on the support to cause irreversible protein adsorption.

Wide Applicability
PRP-3 columns are easy to use and ideal for almost any protein separation, including pH conditions from 1 to 13, with organic modifier (0-100%) of virtually any type from routine to the unusual.  

Scaleup
A selection of column sizes are available from analytical to semi-prep and preparative scale. Sample scaleup is easy because the PRP-3 packing is consistent from analytical to preparative columns. This saves time and eliminates the need to redevelop your separation on semi-prep or preparative columns. The short analytical (50 mm) column is well suited for the gradient elution of high molecular weight proteins (see Application #251). The longer (150 mm) column is well suited for smaller proteins (see Application #252).  

Polymeric columns for protein separations

• Hamilton HPLC Column Instructions

• Hamilton HPLC Column Compound Index

• Column Restoration Procedure

• HPLC Resin Specifications