CM Seplife® FF Agarose Chromatography Resin
Agarose chromatography Resin
CM seplife FF
CM Seplife® FF Agarose Chromatography Resin Product profile:
CM seplife® FF chromatography medium is a new type of highly cross-linked agarose chromatography medium independently developed by Sunresin. It is a weak cation formed by bonding carboxymethyl groups to agarose gel filtration chromatography medium. Ion exchange chromatography media. It has high flow rate, low back pressure, high dynamic load, good chemical stability and mechanical properties, low non-specific adsorption, high recovery rate, easy to scale up, can shorten production time and improve production efficiency. Widely used in ion exchange chromatography purification of downstream proteins, nucleic acids and peptides in biopharmaceuticals and bioengineering.
CM Seplife® FF Agarose Chromatography Resin Parameters:
| Resin code | CM Seplife® FF |
| Appearance | White Sphere beads |
| Particle size(μm) | 45~165 |
| Matrix | Seplife 6FF |
| Flow rate(cm/h) | <750 |
| Pressure(MPa) | 0.3 |
| Heat resistance | 121 ℃, in water for 30 mins |
| pH stability | 4~13 (long term), 3~14 (Short term, CIP) |
| Application |
Proteins, nucleic acids and peptides downstream of biopharmaceuticals and bioengineering |
| Chemical stability |
Stable in the following liquids: all common aqueous buffers; 1 mol/L sodium hydroxide; 8 mol/L urea; 8 mol/L guanidine hydrochloride; 70% ethanol |
| Adsorption (mg / ml) | 60mg/ml lysozyme/ml |
| Ion exchange capacity | 0.09～0.13mmol /ml |
| Working temp. | 4~40℃ |
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Test conditions: Chromatography column 16mm * 300mm; Column bed height 15 cm; temperature 25℃; mobile phase was 0.1 mol / L NaCl. |
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CM Seplife® FF Agarose Chromatography Resin Test Instruction:
1.Column packing
Packing operation in accordance with standard operating procedures. It must be ensured that each material is at working temperature, and the gel needs to be degassed before packing.
2.Balance
Equilibrate the column with 2 to 5 times the bed volume of the loading balance solution. Make sure that the conductivity and pH of the effluent are exactly the same as the conductivity and pH of the loading buffer. The balance solution is a low-concentration buffer solution, such as Tris, PBS, etc.
3.loading
(1)The sample is prepared with the balance solution, and the turbid sample should be centrifuged and filtered before loading. Samples with too much salt concentration are prepared after processing.
(2)Generally, the target product is bound to the column, the impurities are washed away with the equilibration solution, and then the eluent is selected to wash the target product.
(3)The degree to which the medium adsorbs the sample components depends on the charged properties of the sample, the ionic strength of the mobile phase, and the pH value. The salt concentration is small, and the medium adsorbs the sample components more firmly. When using CM chromatography media, the recommended pH value is 1 unit greater than the isoelectric point of the target product.
4.Elution
Increase the salt concentration or increase the pH value for elution, commonly used to increase the salt concentration.
5.Regeneration
Generally, first wash with a high salt concentration buffer (containing 1~2mol/L NaCl) or reduce the pH by more than 10 times the column volume, and then wash to equilibrium with the equilibration solution when binding protein.
If there are inactivated proteins or lipids that cannot be washed away during regeneration, they can be removed with CIP.
6.Cleaning in place
(1)For the protein bound by ionic bonding, it can be removed by 0.5~1 times the bed volume of 2M NaCl.
(2)For precipitated proteins, hydrophobically bound proteins or lipids, you can first rinse with 1M bed volume of 0.1M NaOH, and then wash with equilibration buffer solution until the pH is neutral.
(3)For proteins and lipids with strong hydrophobic binding, wash with 4 to 10 times the bed volume of 70% ethanol or 30% isopropyl alcohol. It should be noted that the concentration of organic solvent gradually increases in a gradient manner, otherwise it is easy Bubbles are generated.
7.Storage
Sealed and stored at 4~30℃(preservation solution is 20% ethanol) in a dry, ventilated, clean place, and can not be frozen; used columns are stored at 4~8℃, 20% ethanol solution.
8.Transportation
Avoid sunlight, rain and heavy pressure during transportation, and it is strictly forbidden to transport with toxic and harmful materials.
CM Seplife® FF Agarose Chromatography Resin Matters needing attention:
(1)Column selection: In theory, as long as the column is long enough, you can get the ideal resolution, but because the column flow rate is related to the pressure gradient, increasing the column length makes the flow rate slower, the peak broadens, and resolution Decrease; the diameter of the column increases, so that the unevenness of the liquid flow increases, and the resolution decreases significantly. A
(2)The pH and ionic strength of the elution buffer must be strictly controlled during the purification process. The sample and CM chromatography medium must be thoroughly equilibrated with elution buffer before column chromatography. A
(3)The installed column bed must be flat and free of channels and bubbles, otherwise it should be reinstalled.
(4)The flow rate should be strictly controlled during the elution, and not too fast.
(5)The volume of the sample should be small, and the concentration should not be too high.
(6)Prevent the cylinder from drying out during sample addition and the entire elution process.
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