Phenyl Focurose FF(HS) is a hydrophobic ligand coupled to provide interactions with hydrophobic groups on the surface of proteins or antibodies, particularly under high ionic strength conditions (where increased ionic strength enhances the interaction between ligands and hydrophobic groups). This enables the separation and purification of biomolecules. Phenyl Focurose FF(HS) is primarily used for initial sample capture and moderate purification.
Features
Fast and simple (one-step purification)
Compared to reversed-phase chromatography, the ligand concentration on the hydrophobic interaction resin is low, leading to mild elution conditions that help preserve the biological activity of molecules
Versatile application: it can be used for initial capture and moderate purification individually, or in combination with ion exchange resins
High loading capacity
Phenyl Focurose FF(HS) Performance Parameters
Resin | Highly cross-linked 6% agarose |
Particle size range | 45-165µm |
Average particle size (D50) | 90±5µm |
pH stability | 2-14 (long-term) 3-13 (short-term) |
Chemical stability | Compatible with all common buffer solutions, 1M acetic acid, 1M sodium hydroxide, 8M urea, 6M guanidine hydrochloride, 30% isopropanol, and 70% ethanol |
Linear flow velocity (0.3 MPa) | ≥300 cm/h |
Maximum pressure | ≤0.3MPa |
Storage solution | 20% ethanol |
Storage conditions | 4℃-30℃ |
Factors affecting hydrophobic interation resins
Influencing Factors | Mechanism | Recommendations |
Ligand structure | Different ligands have varying binding affinities for proteins | It is advisable to perform pre-experiments to select the appropriate resin |
Ligand concentration | Higher ligand concentrations result in stronger binding affinity | Pre-experiments should be conducted to determine the optimal ligand concentration |
Sample properties | The hydrophobicity of a protein depends on the distribution of hydrophobic groups on its surface | / |
Salt concentration | Higher salt concentrations lead to stronger binding between the ligand and the protein, but excessively high salt concentrations can cause protein precipitation | It is important to evaluate protein solubility and stability under different salt concentrations |
Salt type | Different types of salts can yield different binding effects | Priority should be given to (NH4)2SO4 and NaCl |
Temperature | Higher temperatures enhance protein hydrophobicity | It is crucial to maintain a consistent temperature, preferably at room temperature |
pH | Extreme pH values can affect protein solubility and stability, as well as binding efficiency | It is recommended to maintain the pH within the range of 5.0-8.5 while ensuring protein solubility and stability |
Frequently Asked Questions and Solutions
Issue | Possible Causes | Solutions |
Target protein does not bind or has low binding capacity during purification | 1. Overloading of sample | Reduce the sample load |
2. Sample flow rate is too fast | Lower the sample flow rate | |
3. Aggregation of impurities proteins or lipids in the resin | Clean the resin promptly and effectively or replace it with a new resin | |
4. Lower salt concentration in the equilibration buffer or weaker hydrophobicity of the target molecule | Increase the salt concentration in the equilibration buffer or change the type of salt, or replace it with a hydrophobic resin with stronger binding capacity | |
Not collecting the target protein during elution or collecting only a small amount of the target protein | 1. Target protein does not bind to the resin or has low binding capacity | First, confirm if the target protein binds to the resin |
2. Insufficient elution time | Reduce the flow rate to extend the retention time of the elution buffer | |
3. Insufficient elution volume | Increase the elution volume | |
4. Excessive binding strength between the target molecule and the resin | Reduce the salt concentration in the equilibration buffer or change the type of salt, or replace the hydrophobic resin with weaker binding capacity. Add additives (such as a small amount of detergent or low concentration organic reagents) to the elution buffer | |
Target protein purity is low | 1. Sample not pre-processed | The sample must be centrifuged or filtered before loading onto the column |
2. The sample has high viscosity | Dilute the sample with an appropriate equilibration buffer to reduce viscosity | |
3. Incomplete removal of impurities | Increase the washing volume until the baseline stabilizes and matches the equilibration buffer | |
4. Impurities such as proteins or lipids aggregate and precipitate in the resin | Clean the resin promptly and effectively | |
5. Poor elution conditions, such as excessive elution flow rate or steep gradient | Optimize the elution conditions | |
6. Poor column packing | Repack the column or purchase a new one | |
7. Large sample volume retained at the top of the column | Repack the column or reduce the sample volume | |
8. Inappropriate selectivity of the resin | Screen and select an appropriate hydrophobic resin | |
9. Microbial growth in the resin | After using the resin, store it correctly and promptly to prevent microbial growth. | |
Decrease in resin loading | 1. Too fast sample flow rate | Reduce the sample flow rate |
2. Aggregation of proteins or lipids in the resin resulting in decreased loading |
Clean the resin promptly | |
3. Excessive use | Replace with a new resin | |
Rapid increase in chromatographic peak | Overly tight packing of the resin | Repack the column |
Slow or tailing chromatographic peak | Loose packing of the resin | Repack the column |
Cracks or dryness in the column bed | Leakage or introduction of large air bubbles | Check for leaks or bubbles in the tubing and repack the column if necessary |
Slow liquid flow | 1. Aggregation of proteins or lipids | Clean the resin or membrane promptly |
2. Protein precipitation in the resin | Adjust the composition of the equilibration and elution buffers to maintain the stability of the target molecule | |
3. Microbial growth in the separation column | All reagents used must be filtered and degassed. The sample must be centrifuged or filtered before applying it to the column |
Order information
Product | Specification | Item number |
Phenyl Focurose FF(HS) | 25mL | HS060302025M |
Phenyl Focurose FF(HS) | 100mL | HS060302100M |
Phenyl Focurose FF(HS) | 500mL | HS060302500M |
Phenyl Focurose FF(HS) | 1L | HS060302001L |
Phenyl Focurose FF(HS) | 5L | HS060302005L |
Phenyl Focurose FF(HS) | 20L | HS060302020L |
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