Ceramic HyperD® Ion Exchange Chromatography Sorbents (Q, S, DEAE, and CM)
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Preparative Sorbents for the Purification
of Biomolecules by Charge
- High binding capacity. “Gel-in-a-shell” design delivers outstanding dynamic capacity and exceptional dimensional stability for unsurpassed productivity.
- High flow rates. Proteins diffuse rapidly within the hydrogel, facilitating rapid uptake of product. This mechanism of mass transfer (known as “enhanced diffusion”) allows the sorbents to operate free of constraints.
- Abundant ion exchange sites in the hydrogel are highly accessible to protein molecules.
- Does not shrink or swell in response to changes in pH, ionic strength, or flow rate.
- Rigid, non-compressible sorbents are easy to pack.
- Easy cleaning with sodium hydroxide.
- Scalable from research and development to manufacturing.
- Direct capture of biomolecules from a variety of feedstocks
- Purification of polypeptides, IgG, and albumin
- Large-scale purifications
- Purification of monoclonal antibodies from ascites or cell culture
- Plasmid purification
- Process polishing steps
- Rapid, high resolution purification (20 µm grade)
- Ceramic HyperD 20 particle size is more adapted to polishing steps or rapid separations when a higher resolution is required
| Type of Ceramic HyperD Resin | Q | S | Q | S | DEAE | CM |
| Grade | 20 | 20 | F | F | F | F |
| Particle Size (µm) | ~20 | ~20 | ~50 | ~50 | ~50 | ~50 |
| Dynamic Binding Capacity (mg/mL) 10% Breakthrough at 200 cm/h |
BSA 851 | Lysozyme 852 | BSA 851 | Lysozyme 852 | BSA 851 | IgG 603 |
| Amount of Ionic Groups (µeq/mL) | 250 | 150 | 250 | 150 | 200 | 250 - 400 |
| Working pH | 2 - 12 | 2 - 12 | 2 - 12 | 2 - 12 | 2 - 12 | 2 - 12 |
| Cleaning pH | 1 - 14 | 1 - 14 | 1 - 14 | 1 - 14 | 1 - 14 | 1 - 14 |
| Volume Changes Due to pH and Ionic Strength |
Non -compressible | |||||
| Pressure Resistance | 20 grade: 200 bar (20,000 kPa, 2,901 psi) | F grade: > 70 bar (7,000 kPa, 1,015 psi) | ||||
| Storage Temperature |
2 - 30 °C (36 - 56 °F) | |||||
1Sample: 5 mg/mL BSA in 50 mM Tris-HCl buffer, pH 8.6
2Sample: 5 mg/mL lysozyme in 50 mM sodium acetate, pH 4.5
3Sample: 5 mg/mL Human IgG in 50 mM sodium acetate, 100 mM NaCl, pH 4.7
Dynamic Binding Capacity of Ceramic HyperD Ion Exchange Resin Packed in 1 mL Chromatography Glass Columns at a Range of Flow Rates (Linear Velocities)
| Dynamic Binding Capacity (mg/mL)1 | |||
| Media | 1 mL/min (258 cm/h) | 5 mL/min (1290 cm/h) | 10 mL/min |
| HyperD Q-20 µm | 106.0 | 91.5 | 82.5 |
| HyperD F DEAE | 101.5 | 87.5 | 77.5 |
| HyperD F S | 80.5 | 61.5 | 53.5 |
| HyperD S-20 | 97.0 | 89.5 | 83.5 |
| HyperD F CM | 108.0 | 87.5 | 73.5 |
Impact of Linear Velocity on Dynamic Binding with HyperD Ion Exchange Resin
Panel A, Anion Ion Exchange
Ceramic HyperD ion exhange resins (1mL) were packed into Omnifit glass columns (6.6 mm diameter x 2.8 cm bed height) and equilibrated with 25 nM Tris HCl pH 8.5 (anion); and 10 mM MES-NaOH pH 5.8 (cation) at 1 mL/min until a stable pH and conductivity were obtained. Solutions of 5 mg/mL BSA and lysozyme were then pumped onto their respective anion or cation columns at 1 mL/min until "break through" was seen on the absorbance trace at 280 nm.
Panel B, Cation Ion Exchange
The protein solution pumping continued until a plateau of absorbance was seen, usually after 15 CV. The dynamic binding capacity was then calculated at 10% of the plateau value, allowing for system dead volume and expressed as mg/mL of media. This study was repeated at high flow reates of 5 and 10 mL/min.
1Dynamic binding capacity measured by breakthrough curve analysis at 10% of media saturation; a 1 mL volume column of ion exchange resin was packed and equilibrated with 25 mM Tris HCl pH 8.5 (anion ion exchange) or 10 mM MES-NaOH pH 5.5 (cation ion exchange) at the flow rates of 1, 5, or 10 mL/min. For anion ion exchange, 5 mg/mL BSA in the above buffer was then pumped onto the column until a break through in absorbance at 280 nm was seen. The flow was continued until a plateau in absorbance was achieved corresponding to 100% protein feed. Dynamic binding capacity was then calculated at 10% of the plateau value, correcting for any "dead volume" in the system and expressed as mg BSA/mL media volume. For cation ion exchange, 5 mg/mL lysozyme was used to test these resins in a similar manner to the anion ion exchange media.
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