Seplife® LXMS 15S Ion Exchange Resin (DVB)

Synthetic Chromatography Resins

Appearance: White to light yellow spherical beads
Type: Strong acid cation-Sulfonic acid
Matrix: Polystyrene/divinylbenzene
Ligand: Sulfonic acid
Ion exchange capacity (mmol/ml): 0.07-0.10
Particle size range (μm): 13-17
Pore ​​size (nm): 500
pH stability: 2-12 (operation), 1-14 (CIP)
Chemical stability: Stable in commonly used aqueous ion exchange buffers
Flow rate* (cm/h): Max 600
Dynamic binding capacity** (mg/ml): ≥90
Shipped as: Slurry in 20% ethanol solution

*Testing conditions: chromatography column 16mm×200mm; Column bed height 100mm; Packing pressure 2 MPa; Mobile phase 0.5MNaCl.
**Testing conditions: Chromatography column 8mm×100mm; Column bed height 100mm; Packing pressure 0.5 MPa; Mobile phase: 20mmol/L PB buffer pH 6.8; Sampe: Lysozyme; Retention time 2 minutes.

Column loading
The best column packing results are obtained with the resin slurry in 0.5M NaCl at a concentration of 60%-70%.
· Mix the resins to form a homogenate and measure the desired mass or volume, about 1.2 times the column volume.
· Replace 20% ethanol with 0.5 M NaCl solution and equilibrate overnight.
· Before loading the column, adjust the slurry concentration to 65-70% with 0.5 M NaCl solution; pour the resin slurry into the chromatography column in one go, allow to sediment by gravity and mark the height after sedimentation.
· Load the distributor and adjust the height so that the compression coefficient is 1.05-1.10; then pump 0.5M NaCl through the column and stabilize the bed with 1.5 to 2 times the highest working flow rate. (2 - 5 column volumes).
· Efficiency and asymmetry determinations can be performed as described below.

Column Efficiency Evaluation
The test method for column efficiency of the chromatography columns is as follows:
Mobile phase: 0.5 M NaCl solution
Linear flow rate: 100 cm/h
Sample: 2 M NaCl solution
Loading volume: 0.5-1% of column volume
Detection: conductivity detector
Acceptable column asymmetry factor is in the range 0.8-1.8.

Rinsing
The loaded columns should be rinsed with at least 5 BV of deionized water.

Equilibration
Equilibrate the column with an appropriate 5-10 column volume buffer until the conductivity and pH of the effluent remain unchanged.

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
After loading the sample, continue rinsing with equilibration buffer until the baseline is stable. Typically, the molecules retained on the chromatographic media are eluted by increasing the salt concentration or changing the pH to ensure repulsion of the charged species. Occasionally, a solution containing a small percentage organic solvent such as acetonitrile or ethanol is required to disrupt any hydrophobic interactions.

Regeneration and CIP
Regular CIP can prevent the enrichment of protein precipitation impurities in the column bed, and help to maintain the capacity and separation effect of the chromatography media. In general, specific CIP methods and the frequency of CIP need to be designed for each process according to the type of contaminants. The recommended regeneration and CIP method is as follows: Rinse with 5 BV 1-2 M NaCl followed by 5 BV 0.5-1 M NaOH.

Storage
Sealed and stored at 4-30°C (storage solution is 20% ethanol) in a ventilated, dry and clean place, do not freeze.