Seplife® RP LX-21SS (L) Reversed Phase Resin (DVB)

Synthetic Chromatography Resins

Appearance: White spherical beads
Matrix: Poly divinylbenzene
Particle size (μm): 100-300
Uniformity Coefficient: ≤1.3
pH Limits, Stability: 1-14
Pore ​​Volume: 1.70±0.40
Specific Surface Area (m2/g): ≥600
Pore ​​Radius (Å): 150-500
Shipped as: 20% Ethanol Slurry
Flow rate (cm/h): > 400 cm/h, 0.3MPa (0.1M NaCl)
Recommended operating temperature (°C): 20% Ethanol Slurry
Shipped as: Slurry in 20% ethanol solution

The Seplife® RP LX-21SS group of resins can be used in chromatographic columns in a packed bed or in batch mode. When using in column packed bed, the resins can withstand pressures up to 30bar for the Seplife® RP LX-21SS(S) and Seplife® RP LX-21SS(M) and up to 20bar for Seplife® RP LX-21SS(L).
Suggested applications are:
・Purification of small peptides, oligonucleotides and proteins
・Adsorption of vitamins, antibiotics, enzymes, steroids and other substances from fermentation broth
・Decolorization of sugar solutions
・Adsorption of fatty acids
・Adsorption of perfumes
・Decolorization and purification of various chemicals
(6) Lock the column plunger in the compressed position so that the column can be operated in the Static Axial Compression (SAC) mode.
(7) The packed column is now ready for use. It can be used while still assembled on the packing station or it can be undocked for use in a purification facility.

Column Efficiency Evaluation
After packing, clean the chromatographic column with 3-5 CV of 100% methanol or 80% acetonitrile solution. The flow rate should be controlled at 120-180 cm/h to balance and perform column efficiency test.
The test method for column efficiency of RP chromatography columns is as follows:
Mobile phase: 100% methanol or 80% acetonitrile solution
Linear flow rate: 120-180 cm/h
Sample: 1:9 (v:v) Acetone in 100% methanol or 80% acetonitrile
Loading volume: 1% of column volume;
Detection: UV @ 254 or 280 nm;
The prep-HPLC system geometry, including dead volume, will significantly affect the plate count determination.

Equilibration
Equilibrate with the mobile phase for 3-4 CV, and control the flow rate at 120-180 cm/h until the conductance and pH of the flow-through remain unchanged before loading the sample.

Sample feeding
The solid sample can be prepared by dissolving in the equilibrium solution. Low-concentration sample solutions can be concentrated in advance as much as possible while too high concentration sample solution can be diluted with the equilibration solution. To avoid clogging of the column, samples should be processed by centrifugation or membrane filtration. The feed amount is calculated according to the capacity of the resin and the content of the target protein in the feed solution. Before loading, ensure the sample buffer is as consistent as possible with the equilibration solution.

Elution
Use 2-10 CV of methanol, ethanol, acetonitrile, acetone, etc. (aqueous) solution to elute; use acid, caustic or buffer to adjust the pH or use a combination both to elute the molecules of interest.

Regeneration and CIP
First use acetonitrile, methanol, ethanol, acetone, NaOH in ethanol or other solvents to wash the column (3-4 CV) according to the operating flow rate, and then use the equilibration solution to rinse (3-4 CV).

Storage
Chromatography resins that are not for immediate use should be stored in 20% ethanol solution at 4-30 °C.Seplife® RP LX-21SS(S) and Seplife® RP LX-21SS(M) are shipped in 20% ethanol and Seplife® RP LX-21SS(L) is shipped in wet form.