Hamilton’s PRP-3 is a polymeric reversed-phase HPLC column designed for the purification and isolation of proteins and peptides with very good recovery (> 90%). It is based off of the PRP-1 but utilizes a 300 Å pore size rather than the PRP-1’s 100 Å pore size. The highly inert polymeric packing poly(styrene-divinylbenzene) enhances protein recovery because there are no silanol groups on the support to cause irreversible protein adsorption.
PRP-3 is a PS-DVB support that is pressure stable up to 5,000 psi and cross-linked to prevent shrinking or swelling when the mobile phase is changed. Chemically, proteins present solubility problems unlike many small molecules. Most proteins are hydrophobic on the inside, with highly charged exteriors. This often presents dissolution problems, particularly when pH is near the isoelectric point of the protein. The rugged chemical nature of the PRP-3 allows the protein chemist a much broader selection of agents for dissolution, including concentrated acids, aggressive chaotropes, as well as detergents. In proteomics, the PRP-3 has excellent potential for single-column 2D HPLC. The orthogonal selectivity between low and high pH separations is often comparable to that achieved with two different column formats (SCX, RP), but with the added bonus of MS-compatible mobile phase.
PRP-3 stationary phase structure and applications
Organic compounds: large molecules (> 2,000 mw), peptides, proteins, protein digests, protected and de-protected oligonucleotides, nucleic acids
Examples of analytes that can be separated on PRP-3 columns:
- Globular proteins
- Antibody fragments
- Tryptic digests
- RNA oligomers
- Synthetic high mw polymers