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Specification Sheet: Method Development Guide for Hypercarb columns Product Literature

Application Note: An Improved UHPLC-MS/MS Assay for the Determination of Acrylamide from Foodstuffs Product Literature

Specification Sheet: Preparative HPLC Columns – Scale up to More Options Product Literature

Brochure: Preparative HPLC Columns – Scale up to More Options Product Literature

Specification Sheet: Syncronis C18 HPLC Columns Product Literature

Application Note: Analysis of Ibuprofen and Valerophenone Using an Accucore XL C18 4 μm HPLC Column Product Literature

Application Note: Automated Extraction and Determination of Human Hormones in Drinking Water Using Solid-Phase Extraction and HPLC with UV Detection Product Literature

Technical Bulletin: Comparison of the Chromatographic Resolution of Solid Core 4 μm and Fully Porous 3 μm and 5 μm Columns Product Literature

Are the pre-packed POROS CaptureSelect or any POROS analytical columns compatible with an UPLC or is the use of these columns limited to HPLC? Product FAQ

Answer

UPLC is typically described as chromatography with sub-2 µm particles, which drives higher efficiency/better separation independent of operating flow rate. As particle size decreases, backpressure increases so the UPLC systems tend to have higher pressure ratings compared to HPLC systems. The flow rates on UPLC systems would be sufficient for POROS columns. Pump capabilities are typically 0.1-1.0 mL/min, so not as fast as a HPLC, but sufficient. Hold-up volumes on UPLC systems are lower than HPLCs, so this can help with sensitivity and efficiency. So from a pressure and flow standpoint there should be no challenge to operate a POROS CaptureSelect column on a UPLC system.

That being said, care should be taken running POROS columns on UPLC systems. Higher pressure safety interlock MUST be set at the rated column pressure, typically 180 bar. HPLC column hardware is not rated to operate at the static pressures possible and typical on UPLC systems. A clog or the like on a UPLC system could generate unsafe pressures on any HPLC-rated column before the default UPLC safety pressure interlock is tripped.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Answer Id: E12922

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Are POROS analytical columns compatible with HPLC systems from Waters or Agilent? Product FAQ

Answer

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.

Answer Id: E12924

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Can POROS resins be used to purify large biomolecules? Product FAQ

Answer

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.

Answer Id: E12916

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How many uses should I expect out of my POROS or CaptureSelect column? Product FAQ

Answer

Resin lifetime depends on how the resin is used and the cleaning process that is employed. Therefore, each purification process will need to be evaluated specifically, especially if long lifetime is desired. Because binding can be different between resins, different cleaning schemes may be needed. An unoptimized cleaning process of any resin may yield <5 uses, and an optimized cleaning process can yield a hundred cycles or more. Discoloration of resin can occur for different reasons from process compounds such as metals like iron (Fe2+/3+) and copper (Cu2+), additives to the media (like vitamin B12 and folic acid), and elution solutions (like magnesium chloride, Mg2+). We can provide recommendations that have proved helpful for many customers to optimize the life of their POROS or CaptureSelect column.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Answer Id: E12915

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What type of column do you recommend for POROS 50 µm resins? Do I need a high pressure column for POROS resins? Product FAQ

Answer

POROS resins are mechanically rigid and incompressible and can be packed effectively in low-pressure glass columns and in high-pressure stainless steel columns. The lack of wall support with increasing column diameter has minimal impact on chromatography performance because the beads support themselves, allowing for flexible column packing approaches and consistent and robust results. Columns can be packed with traditional flow pack, axial compression, or pack-in-place/stall pack packing methods.

Find additional tips, troubleshooting help, and resources within our Protein Purification and Isolation Support Center.

Answer Id: E12914

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For any POROS analytical column, is the column compatible with an AKTA or Avant purification system? Product FAQ

Answer

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.

Answer Id: E12923

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What is the physical or mechanical stability of POROS resins? Product FAQ

Answer

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.

Answer Id: E12920

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