POROS™ CaptureSelect™ LC Lambda Affinity Column, 4.6 x 50 mm

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.

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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.

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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.

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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.

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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.