Hydrocell
|
| |||||||||||||||||||||||||||||||||||||||||||
Ion Exchange HPLC Columns HYDROCELL ion exchange adsorbents were developed by BioChrom Labs, Inc. for protein and other biomolecule separations. They are based on the coatings of the neutral hydrophilic layer on the rigid macroporous polystyrene-divinylbenzene (PS-DVB) matrix, followed by chemical immobolization of the polymer chains of bonded phase on the hydrophilic surface. The packings do not have any weak ester or amide linkages as do the other polymeric gels. Their chemical and physical stabilities allow rapid equilibration and fast clean-up using high salt, HCl, NaOH and organic solvents. Unlike some columns where hydrophobic retention can complicate ion exchange chromatography by producing irreversible adsorption, HYDROCELL ion exchangers are very hydrophilic. Elution of proteins can be achieved with buffers of moderate ionic strength, typically less than 0.5M NaCl, in 15 minutes at a flow rate of 1.0 ml / minute. |
| |||||||||||||||||||||||||||||||||||||||||||
Weak Anion Exchanger: DEAE 1500 and DEAE 3000 DEAE 1500 is prepared from 10µm particles of highly cross-linked PS-DVB beads with pore diameter of 500 Å. Similarly, DEAE 3000 is prepared from 10 µm particles of PS-DVB beads with average pore diameter of larger than 1000Å. The packing has an optimum pore size and is suitable for loading and analysis of large globular proteins. In coupling of the polymer chains of diethylaminoethyl (DEAE) groups with the hydrophilic surface of the rigid macroporous polymer matrix, weak anion separations can be achieved over the wide pH range of 1 to 13. Elution is accomplished with gradients of increasing ionic strength or pH. The dynamic loading capacity for BSA is about 30 mg/ml of column volume. Recoveries of proteins are typically from 95% to quantitative. Chromatogram for Protein Mixture on DEAE 1000 Column |
| |||||||||||||||||||||||||||||||||||||||||||
Strong Anion Exchanger: QA 1500 and QA 3000 QA 1500 and QA 3000 are quaternary amine anion exchanger designed for the rapid separation of proteins and enzymes. The rigid macroporous PS-DVB matrix with a hydrophilic surface has been bonded with the polymer chains of diethylmethylaminoethyl groups to provide the optimum density of strong anion exchange functionalities for macromolecule separations. The media is compatible with aqueous solvents in the pH range of 1 to 13 and with most organic solvents. It provides a mild, nondenaturing adsorbent from which proteins can be eluted with buffers of moderate ionic strength (< 0.5M NaCl) in 15 minutes at a flow rate of 1.0 ml/minute. HYDROCELL anion exchangers are very hydrophilic, proteins with pI close to or above the mobile phase pH will not be retained under these conditions. Chromatogram for QA 1000 Column
|
| |||||||||||||||||||||||||||||||||||||||||||
Weak Cation Exchanger: CM 1500 and CM 3000 CM 1500 is prepared from 10 µm particles of highly cross-linked PS-DVB beads with pore diameter of 500 Å. Similarly, CM 3000 is prepared from 10 µm particles of highly cross-linked PS-DVB beads with pore diameter of larger than 1000 Å. The polymer has been completely covered with neutral, hydrophilic layer to shield the hydrophobic surface. In coupling of the polymer chains of carboxymethyl (CM) groups with the rigid macroporous polymer matrix, weak cation separations can be achieved over the wide pH range of 2 to 13. The separation is usually accomplished with salts or pH gradients using an aqueous mobile phase. The dynamic loading capacity for lysozyme is ca. 30 mg/mL of column volume. Recoveries of proteins are typically from 95% to quantitative. Chromatogram of Protein Mixture on CM 1000 column |
| |||||||||||||||||||||||||||||||||||||||||||
Strong Cation Exchanger: SP 1500 and SP 3000 SP 1500 is prepared from 10 µm particles of highly cross-linked PS-DVB beads with pore size of 500 Å. Similarly, SP 3000 is produced from 10 µm particles of highly cross-linked PS-DVB beads with pore size of larger than 1000 Å. After being coated with neutral, hydrophilic layer, the polymer is chemically bonded with the polymer chains of sulfopropyl (SP) groups to provide the optimum density of strong cation exchange functionalities for macromolecule separations. The media is compatible with aqueous solvents in the pH range of 1 to 13, as well as with most organic solvents. Elution of proteins can be achieved at relatively low ionic strength (< 0.5 M NaCl) in 15 minutes at a flow rate of 1.0 mL/min. Because of the hydrophilic nature of the adsorbents, proteins with pI value close to or below the mobile phase pH will not be retained on Hydrocell cation exchange columns. Chromatogram of Protein Mixture on SP 1000 |
| |||||||||||||||||||||||||||||||||||||||||||
| ||||||||||||||||||||||||||||||||||||||||||||
Recovery of Proteins from HYDROCELL Ion Exchange Columns |
| |||||||||||||||||||||||||||||||||||||||||||
Non-Porous High Speed Ion Exchange Columns | ||||||||||||||||||||||||||||||||||||||||||||
