Are POROS analytical columns compatible with HPLC systems from Waters or Agilent?
All of the pre-packed POROS columns, regardless of column size or resin, have the same 10-32 female fittings on the column. Columns are compatible with Waters and Agilent HPLC systems. You will need 1/16 inch male fittings for the HPLC to attach the column.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
For any POROS analytical column, is the column compatible with an AKTA or Avant purification system?
All of the pre-packed columns, regardless column size or resin, have the same 10-32 female fittings on the column. They are all compatible with the AKTA instrument platform, however due to the column volume of the 2.1 mmD x 30 mmL columns (0.1 mL), these will not perform well in AKTA or Avant purification systems due to the relative size of the hold- up volume in these systems. The smallest column (PEEK or SS) that could feasibly be used in these systems is the 4.6mmD x 50mmL (0.8 mL) format.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
What is the physical or mechanical stability of POROS resins?
The physical stability of POROS resins is high. Customers have agitated, with overhead mixing and or bottom gassing for long periods of time (5-10 hours) and have performed hundreds of diaphragm pump passes during stall packs. Keep in mind that agitation with any device that creates grinding and sheer, such as with a magnetic stir bar, is not recommended for any type of resin.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
What should I do if a POROS column dries out or air is pumped onto the column?
POROS resins rehydrate quite well. Here is the procedure that we recommend:
1. Flow 5% ethanol over the column at ?350 cm/hour for 3 CVs, followed by water or 0.1 M sodium chloride at 300 cm/hour for 3 CVs.
2. If baseline noise is still present, flow 20% ethanol over the column at 300 cm/hour for 3 CVs, followed by water or 0.1 M sodium chloride at 300 cm/hour for 3 CVs.
3. If you have established column qualification tests and specifications, perform the tests after rewetting.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
Can POROS resins be used to purify large biomolecules?
In general POROS resins have behaved better for large biomolecules such as viral particles, fusion proteins, and globular proteins, where shape (not size) matters due to its unique pore structure and large pore structure as compared to traditional resins. As the target molecule size increases, capacities obtained will decrease. The large pore structure of POROS resins, which allows for convective flow (and therefore enhanced diffusion) is especially well suited for the purification of large biomolecules. POROS resins offer the best blend of high capacity (associated with chromatography beads) and improved chromatography efficiency (typically associated with monoliths or membranes). The average pore size for POROS resins is 1,000-3,600 Angstrom (100-360 nm) depending on the base bead/chemistry being utilized.
For example, the molecular weight of IgG is 150 kDa and the molecular radius is 55 Angstrom. The molecular weight of IgM is ~900 kDa (pentamer), and the molecular radius is 120 Angstrom. So both of these biomolecules can interact with the pore structure associated with POROS materials.
Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.
