Protein G Focurose 4FF

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℃

Issue

Possible Causes

Solutions

Target protein does not bind or has low binding capacity during purification

Overloading of sample

Reduce the sample load

Sample flow rate is too fast

Reduce the sample flow rate and load the sample according to a retention time of 4-6 minutes

Protein or lipids aggregate in the resin, affecting binding

Efficiently clean the resin or replace with a new resin

Weak binding between target molecule and resin

Confirm the source and subtype of IgG and select the appropriate resin

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.

Target protein does not bind to the resin or has low binding capacity

First, confirm if the target protein binds to the resin

Unsuitable elution conditions

Optimize elution conditions to increase elution strength

Insufficient elution time

Lower the flow rate and extend the retention time of the elution buffer

Elution volume is too small

Increase elution volume

Target protein purity is low

Unreasonable sample pre-processing

Based on the sample source, it is recommended to perform filtering or dilution before column loading

The sample has high viscosity

Dilute the sample appropriately with a balancing solution to reduce sample viscosity and concentration

Incomplete removal of impurities

Gradually increase the wash volume until the baseline stabilizes and matches the equilibrium solution

Impurities such as proteins or lipids aggregate and precipitate in the resin

Clean the resin promptly and effectively

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

Poor column packing

Reload or use new packing material

Large dead volume at the top of the column

Re-pack the column or reduce the dead volume

Microbial growth in the resin

Store the medium properly and promptly after use

Decrease in resin loading

Too fast sample flow rate

Reduce the sample flow rate

Aggregation of proteins or lipids in the resin resulting in decreased loading

Clean the resin promptly

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

Aggregation of proteins or lipids

Clean the resin or membrane promptly

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