Basic operation of ion exchange protein purification system company

The ion exchange protein purification system company is specially designed for the purification and preparation of protein and other biological samples. The high-performance infusion pump not only withstands high pressure, but also has good stability at low temperature and low pressure, providing safety guarantee for low-pressure chromatography columns; using DAD detector , Can facilitate the real-time monitoring of the purity of the separated components. In addition, SCG is equipped with an acidity / conductivity detector to accurately and instantly monitor the gradient environment during separation.

Basic operation of ion exchange protein purification system  

1. Pretreatment of DEAE-cellulose:  

Weigh 2gDE-52 and place in a sand core funnel, first soak in 20mL0.5mol / LNaOH for 30min, then rinse with distilled water to pH = 8.0, then soak in 20mL0.5mol / LHCl for 30min, then rinse with distilled water to pH = 4.0. After zui, soak in 20mL0.5mol / LNaOH for 30min and wash with distilled water to pH = 8.0. Soak the treated DEAE-cellulose with eluent A and replace it 1 to 2 times.

2. Pack a column and take a 1.5ⅹ 20cm chromatography column, as shown in Figure 1. Fill about 10mL of eluent A, open the lower piston to let the buffer drip out slowly, at the same time, pour DEAE-cellulose suspended in the appropriate amount of eluent A into the chromatography column while stirring, let it settle to All have been added. The column was equilibrated with eluent A at a flow rate of 1.0 ml / min until the pH of the effluent was exactly the same as that of the starting buffer.

Note: When loading the column, the exchange agent should be poured once. If it is poured in portions, the exchange agent at the interface must be stirred up before adding again to ensure that the column bed is not divided. The cylinder surface should be flat, and there should be no air bubbles in the column.

3. Loading

After the above balancing process is completed, the piston is closed after the liquid level is about 1 cm from the cellulose sedimentation surface. Use a dropper to carefully add the sample along the wall of the tube. After all the sample enters the column, add a layer of eluent A to the column bed. The sample volume accounts for about 2.5% of the column volume.

4. Elution

Connect the elution bottle and open the bottom of the column to start elution. First elute a column volume with eluent A (collected in a test tube) at a flow rate of about 0.5 ml / min; then elute one column volume with 5 gradients in eluent B and collect separately at a flow rate of about 0.5 ml / min.

5. Identify and collect each eluted component, and determine the protein content of each component by ultraviolet absorption method.

Protein Purification System Company-Suzhou Sepure Instrument Co., Ltd. is committed to developing precision purification systems for biological products such as protein purification, automatic protein chromatography systems, antibodies, natural products, and small molecule drugs. A variety of purification systems with different configurations, from laboratory research level to industrial production amplification level, provide customers with a one-stop shopping experience, and strive to provide customers with stable and practical products and a comprehensive product service system. The company has established good cooperation with many well-known biopharmaceutical companies (including CRO companies) at home and abroad, and the product indicators have reached the international leading level of the same type of products, and has received recognition and praise from many customers. The protein purification system company welcomes everyone to come to consult.