Table 1. Resin Characteristics
|Bead Size||60-160 µm|
|Flow Rate1||>20 mL/min (>680 cm/hr) @ 25oC|
|Storage Temperature||2-8o C|
|Chemical Stability2||Stable in all commonly used aqueous solutions and buffers.|
|Physical Stability2||Negligible volume variation due to changes in pH or ionic strength.|
1Linear flow rate = volumetric flow rate (cm3/h)/column cross-sectional area (cm2)
2Data refer to the coupled product, provided that the ligand can withstand the pH or chemical environment. Please note the following: pH stability, long term refers to the pH interval where the medium is stable over a long period of time without adverse effects on its subsequent chromatographic performance. pH stability, short term refers to the pH interval for regeneration and cleaning procedures.
Instructions for Use
Protein binding to Heparin Superflow is generally most efficient at neutral pH and low ionic strength. Elution usually requires a high ionic strength buffer. Heparin Superflow is manufactured to withstand flow rates of 680 cm/hr. Batch processing is an option that can lead to improved product recovery as well as reduced processing time. As with all chromatography, purification conditions may need optimization.
(Note: buffers for heparin chromatography vary greatly depending on the application; these conditions are recommended for AT3 purification and are not to be considered generic).
1. Measure the appropriate amount of resin to fill the column.
2. Wash the resin with 5 bed volumes (BV) of deionized (DI) water.
3. Equilibrate the resin by washing it thoroughly with 3 BV of Binding Buffer. The recommended flow rate for the washing and equilibrations steps is 200cm/hr.
4. Load protein sample. The recommended flow rate for this step is 100cm/hr.
5. Wash to remove unbound material with excess Binding Buffer. The recommended flow rate for washing is 200cm/hr. The optical density of the wash should approach baseline.
6. Elute bound protein with Elution Buffer until the OD (A280) shows that most of the bound material has been eluted. The recommended flow rate for this step is 100cm/hr.
Binding Buffer: 10mM Citrate, 50mM NaCl, 20% Ethanol, pH 6.8
Elution Buffer: 10mM Imidazole, 4M NaCl, pH 6.5
Storage Buffer: 20% Ethanol
Sanitization and regeneration:
1. Sanitize the resin with 1 BV of 1M NaOH for 1 hour. The maximum recommended flow rate for steps 1-3 is 200cm/hr.
2. Wash the resin with DI water until neutral.
3. Equilibrate the resin by washing with 3 BV of Binding Buffer.
4. Column is now ready for reuse.
Store the cleaned beads in 0.1M NaOH at 2-8oC for up to six months or as a 70% slurry in 20% ethanol for extended storage.
To Download Instructions for use:
INST 37304SF4 Heparin Superflow 4