Seplife® MMC Large Scale/75 Multimodal Strong Base Anion Resin

Multimodal Chromatography - Agarose

Appearance: White to off white spherical beads
Type: Multimodal Weak Acid Cation agarose
Matrix: Highly cross-linked 6% agarose
Ion exchange capacity (mmol/ml): 0.07-0.09
pH ligand fully charged: Negatively charged at pH>5
Particle size range (μm): 45-125
pH stability: 3-12 (operational), 2-14 (CIP)
Chemical Stability: Stable in common aqueous solutions: 1M NaOH, 1M acetic acid. AVOID the use of Oxidizing agents, cationic detergents
Flow rate* (cm/h): max 1000cm/h
10% dynamic binding capacity (mg/ml)**: ≥45
Shipped as: 20% ethanol slurry

*Testing conditions: Chromatography column 16mm×200mm; column bed height 20cm; temperature 25°C; mobile phase water.
** Testing conditions: Binding buffer: 50mM acetate + 0.25M NaCl, pH 4.75; Elution buffer: 50mM acetate + 1.0M NaCl, pH 7.0, Sample: bovine serum albumin, Column: 8mm*100mm, room temperature, Retention time 2 minutes.

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

Equilibration
Equilibrate the column with at least 5 BV of the initial buffer solution until the conductivity and pH of the effluent remain constant. The pH of the initial buffer is 0.5-2.0 pH units lower than the isoelectric point of the target protein.

Sample feeding
Samples are prepared in buffer. Cloudy samples should be centrifuged and filtered before loading.

Elution
Elute with lower conductivity or higher pH buffer. Keep the flow rate and buffer composition unchanged during elution.

Regeneration
Elute the reversibly bound molecules with a solution of high ionic strength (such as 2M NaCl buffer), and adjust the pH to 10-11. Rinse with at least 5 BV of the initial buffer until the conductivity and pH of the effluent remain constant.

Cleaning-in-place (CIP)
1. For proteins bound by ionic bonds, backwash with 0.5-2 BV of 2M NaCl for 10-15 minutes.
2. For precipitated proteins, hydrophobically bound proteins or lipids, backwash with 1M NaOH at a flow rate of 40cm/h for 1 to 2 hours. 3. For proteins and lipids with strong hydrophobic binding, backwash with 2-4 BV of 70% ethanol or 30% isopropanol. However, it should be noted that the concentration of the organic solvent should be gradually increased to avoid bubbles.
After cleaning, equilibrate the column with equilibration buffer solution at least 3 times the volume of the column bed until the pH and conductivity remain unchanged.

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