Seplife® MMC Large Scale/75

Instructions

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. 

Ordering information

Production date: See label

Expiry Date : 5 years, under proper storage conditions