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Seplife®6FF

Product Description

Fast flow agarose chromatography Resin

Seplife 6FF

Product profile:
Seplife® 6FF fast-flow agarose chromatography resin is formed by cross-linking on Seplife® 6B agarose chromatography media. Its chemical stability and physical properties are significantly enhanced with high flow rates, high loading, and low specific adsorption. It has high recovery rate and can be reused many times. It can be used for gel filtration chromatography and purification of proteinsnucleic acids and peptides downstream of biopharmaceuticals and bioengineering.

20200804083702

Resin Parameters:

Resin code Seplife® 6FF
Appearance White Sphere beads
Particle size(μm) 45~165
Matrix 6% cross-linked agarose
Flow rate(cm/h) <300
Pressure(MPa) 0.2
pH stability 2~12 (long term), 2~14 (Short term, CIP)
Application Proteins, nucleic acids and peptides downstream of 
biopharmaceuticals and bioengineering
Chemical stability Stable in water buffer solution: 2M NaOH; 70% ethanol, 30% 
isopropanol, 30% acetonitrile, 1% SDS, 6M guanidine 
hydrochloride, 8M urea.
Exclusion limit 10000~4×106 globulin
Test conditions: Chromatography column 16mm * 300mm; Column bed height 15 cm; temperature 25℃; 
mobile phase was 0.1 mol / L NaCl.


Test Instruction:
1.Column packing
Packing is performed in accordance with standard operating procedures. It must be ensured that each material is at operating temperature and that the gel needs to be degassed before it is packed.
2.Equilibrium
Equilibrate the column with 2 to 5 column volumes of the loading balance, making sure that the conductivity and pH of the effluent are exactly the same as the conductivity and pH of the loading buffer.
3.Sample Loading
(1)The sample is prepared with a buffer, and the turbid sample is centrifuged and filtered for loading. Samples with excessive salt content and too small concentration should be treated first and then loaded.
(2)The separation of the sample components by the medium is carried out according to the molecular weight of the components, and the molecular weight is first flowed out.
(3)The loading volume is about 1-2% of the column volume, and the smaller the better separation performance.
4.Elution
Elution was carried out with buffer, and the flow rate and buffer composition were kept constant during the elution.
5.Regeneration
It is usually washed with a buffer to balance and can be used again. Some inactivated proteins or lipids cannot be washed off during regeneration and can be removed by in-place cleaning (CIP).
6.CIP
(1)For an ion-bonded protein, it can be removed with 0.5 to 1 column bed volume of 2 M NaCl.
(2)For precipitated proteins, hydrophobically bound proteins or lipids, it can be removed with 1 column bed volume of 0.1 M NaOH. (3) For strongly hydrophobically bound proteins, lipids, etc., it can be washed with 4 to 10 column volumes of 70% ethanol or 30% isopropanol. However, it should be noted that the concentration of the organic solvent is gradually increased in a gradient manner, otherwise bubbles are easily generated.
After the cleaning is completed, equilibrate the column with at least 3 bed volumes of equilibration buffer until the pH and conductance remain unchanged.
7.Storage
Sealed and stored at 4~30℃(20% ethanol in storage solution), dry, ventilated, clean, not frozen; used columns are stored in 4℃, 20% ethanol solution.


Caution:
(1)The sample volume should be concentrated as much as possible before using the gel filtration.
(2)The sample must be clarified without particles.
(3)In order to obtain the highest resolution, the loading volume cannot exceed 5% of the bed volume.
(4)Column chromatography should be performed after the sample and the chromatographic medium must be thoroughly equilibrated with the elution buffer.
(5)The column bed must be flat, free of grooves and air bubbles, otherwise it should be reassembled.
(6)In order to protect the stability and activity of the protein, a buffer purification protein is selected.
(7)In order to avoid non-specific ionic interaction between protein and medium, up to 0.15 mol/L NaCl can be added to the buffer.
(8)According to the molecular weight of the target protein, the medium closest to the median separation range is selected.
(9)The flow rate should be strictly controlled during the elution process, and not too fast.
(10)During the loading and the entire elution process, the cylinder surface is prevented from drying out.
(11)This product should avoid contact with oxidizing agents and avoid prolonged exposure to the air.



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