Protein G Focurose 4FF is suitable for the purification of ascites fluid, cell culture supernatants, monoclonal and polyclonal antibodies, and Fc-tagged proteins derived from serum sources.
Features
Fast and simple purification (one-step purification)
High loading capacity, fast flow rate, and easy scale-up
Compared to Protein A Focurose 4FF, Protein G Focurose 4FF has a broader binding surface (for more species sources) and stronger binding capacity
Protein G Focurose 4FF Performance Parameters
Resin | Highly cross-linked 4% agarose |
Particle size range | 45-165µm |
Average particle size (D50) | 90±5µm |
Dynamic binding capacity | ≥20 mg (IgG) /mL |
pH stability | 3-9 (long-term) 2-10 (short-term) |
Linear flow rate (0.3 MPa) | ≥250cm/h |
Operating pressure | ≤0.3MPa |
Storage solution | 20% ethanol |
Storage conditions | 4℃-8℃ |
Note:
1. The binding capacity of the resin may vary depending on the sample source and subtype variations.
2. Prolonged immersion of the resin in the elution solution can result in ligand hydrolysis, ultimately affecting the resin's loading capacity.
Binding Strength of Protein A and Protein G with IgG from Different Species
Species | Subclass | Protein A | Protein G |
Human | IgA | Variable | - |
IgD | - | - | |
IgE | - | - | |
IgG1 | ++++ | ++++ | |
IgG2 | ++++ | ++++ | |
IgG3 | - | ++++ | |
IgG4 | ++++ | ++++ | |
IgM | Variable | - | |
Guinea pig | IgG1 | ++++ | ++ |
IgG2 | ++++ | ++ | |
Hamster | / | + | ++ |
Mouse | IgG1 | + | ++++ |
IgG2a | ++++ | ++++ | |
IgG2b | +++ | +++ | |
IgG3 | ++ | +++ | |
IgM | Variable | - | |
Rat | IgG1 | - | + |
IgG2a | - | ++++ | |
IgG2b | - | ++ | |
IgG3 | + | ++ | |
Rabbit | / | ++++ | +++ |
Egg yolk | IgY | - | - |
Dog | / | ++ | + |
Pig | / | +++ | +++ |
Horse | / | ++ | ++++ |
Cow | / | ++ | ++++ |
Sheep | / | -/+ | ++ |
Goat | / | - | ++ |
Monkey | / | ++++ | ++++ |
Camel | / | - | + |
Bear | / | - | + |
"-" : No binding "+" : Weak binding "++" : Binding "+++" : Moderate binding "++++" : Strong binding |
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 | Reduce the sample flow rate and load the sample according to a retention time of 4-6 minutes | |
3. Protein or lipids aggregate in the resin, affecting binding | Efficiently clean the resin or replace with a new resin | |
4. Weak binding between target molecule and resin | Confirm the source and subtype of IgG and select the appropriate resin | |
5. Inappropriate choice of buffer during purification | Confirm the pH of the sample, and for weakly binding target molecules, increase the pH (8-9) and salt concentration (1-3M NaCl) to enhance the binding between the sample and resin | |
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. Unsuitable elution conditions | Optimize elution conditions to increase elution strength | |
3. Insufficient elution time | Lower the flow rate and extend the retention time of the elution buffer | |
4. Elution volume is too small | Increase elution volume | |
Target protein purity is low | 1. Unreasonable sample pre-processing | Based on the sample source, it is recommended to perform filtering or dilution before column loading |
2. The sample has high viscosity | Dilute the sample appropriately with a balancing solution to reduce sample viscosity and concentration | |
3. Incomplete removal of impurities | Gradually increase the wash volume until the baseline stabilizes and matches the equilibrium solution | |
4. Impurities such as proteins or lipids aggregate and precipitate in the resin | Clean the resin promptly and effectively | |
5. Degradation of the target substance | Pay attention to the storage conditions before and after sample purification, and avoid prolonged exposure to high temperatures, excessive acidity, or alkalinity | |
6. Poor column packing | Reload or use new packing material | |
7. Large dead volume at the top of the column | Re-pack the column or reduce the dead volume | |
8. Microbial growth in the resin | Store the medium properly and promptly after use | |
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. Gradual detachment of ligands | Optimize process conditions and replace with new resin | |
Chromatographic peak rises too sharp | Resin is packed too tightly | Adjust column parameters and repack the column |
chromatographic peak rises too slowly or has a tailing | Resin is packed too loosely | Adjust column parameters and 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. 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 |
Protein G Focurose 4FF | 25mL | HQ030316025M |
Protein G Focurose 4FF | 100mL | HQ030316100M |
Protein G Focurose 4FF | 500mL | HQ030316500M |
Protein G Focurose 4FF | 1L | HQ030316001L |
Protein G Focurose 4FF | 5L | HQ030316005L |
Protein G Focurose 4FF | 20L | HQ030316020L |
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