Seplife® DX 50/40 Dextran-Based Size Exclusion Chromatography Resin

Dextran-Based Chromatography Resins

Appearance: White spherical beads
Type: Gel Filtration
Matrix: Crossed linked dextran
Particle size (dry, µm): 20-80
Swelling property (ml/g): 9-13
pH stability: 3-10 (operational), 2-13 (CIP)
Chemical Stability: Stable in all common aqueous buffers; 1M sodium hydroxide; 8M urea; 6M guanidine hydrochloride; 70% ethanol.
Flow rate* (cm/h): max. 150
Shipped as: dry

*Testing conditions: Chromatography column 16mm×200mm; column bed height 10cm; temperature 25°C; mobile phase water.

Product pretreatment Weigh the required Seplife®DX 50/40 and place it in 50~100 times distilled water for swelling. It usually takes 1~2 days to swell at room temperature, and 2 hours in boiling water to allow the media to fully swell until the volume no longer changes.
Note: Add distilled water before swelling to make it settle naturally. After natural settlement, if there are many fragments floating in the solution, they need to be removed by repeated rinsing to prevent blocking the column during chromatography and reducing the flow rate.

Column packing
Column packing should be performed according to standard operating procedures. It is important to ensure that each material is at its working temperature, and the media may need to be degassed before column packing.

Equilibration
Equilibrate the column with an appropriate 2-5 column volume buffer. Ensure the conductivity and pH of the effluent are exactly the same as the buffer.

Sample feeding
1. The separation of sample components by the media is carried out according to the molecular size of the components, and the ones with the larger molecular size flow through first.
2. The sample feeding for gel filtration is generally 5% of the column bed in volume, and we recommend that the initial sample feeding be controlled at 1-2% of the bed volume, which can be adjusted depending on the separation situation. When desalting, the sample feeding can reach 20% of the column bed volume. The selection of column height is also related to the separation requirements. The column height should be controlled below 40 cm. Too high a gel layer will cause greater back pressure and should be avoided as much as possible.
Difficult-to-separate molecules must have a certain column height and flow rate control. A 5:1 height-to-diameter ratio for desalting is recommended.
3. Samples with particulates and precipitate should be filtered or centrifuged before the chromatography purification. The viscosity of the sample should not be too high, otherwise the separation effect will be reduced.

Elution
Elution can be done with salt-free water or with equilibration buffer as eluent. Complete separation can be achieved by adding NaCl to the equilibration buffer for gradient elution, or salt gradient elution.

Cleaning-in-place (CIP)
After the media is used 10 times, a CIP is performed to remove the precipitated and stubborn residual proteins in the column bed. The method is to backwash with 4 bed volumes of 1M NaOH at 40cm/h, and then regenerate with at least 3 bed volumes of equilibration buffer.

Storage
Seplife®DX 50/40 dry powder should be stored in a dry, ventilated and clean place at 4-30°C; the hydrated media should be stored in 20% ethanol solution. Avoid contact with oxidants and do not freeze.