Application of ultrafiltration in protein separation and purification

Application of ultrafiltration in protein separation and purification1

Ultrafiltration technology is a new and high-efficiency separation technology. It has the characteristics of simple process, high economic benefit, no phase change, large separation coefficient, energy saving, high efficiency, no secondary pollution, continuous operation at room temperature and so on. Today, manager Yang whom from Beijing inquired about our ultrafiltration equipment for protein purification and communicated in detail with our technology. Now, the editor of Shandong Bona group will introduce the application of ultrafiltration in protein separation and purification.

1. For protein desalination, dealcoholization and concentration
The most important applications of ultrafiltration in the purification of proteins are desalting and concentration. Ultrafiltration method to desalination and concentration are characterized by large batch volume, short operation time and high efficiency of protein recovery. The traditional method of steric exclusion chromatography to remove various substances from proteins has been replaced by modern ultrafiltration technology, which has become the main technology for protein desalination, dealcoholization and concentration today. In recent years, ultrafiltration technology has been widely used in the desalination and recovery of high nutritional value proteins in cheese whey and soybean whey. Lactose and salts and other components in the protein, as well as the actual needs of successfully completing the desalting, de-alcoholization and concentration of proteins. The use of ultrafiltration technology can also concentrate serospecies immunoglobulins to meet the actual demand of protein yield.

2. For protein fractionation
Protein fractionation refers to the process of separating each protein component section by section according to the difference of physical and chemical properties (such as relative molecular weight, isoelectric point, hydrophobicity, etc.) of each protein component in the feed liquid. Gel chromatography is one of the most commonly used methods for fractionation of biological macromolecules (especially proteins). Compared with traditional chromatography, ultrafiltration separation technology has a good prospect of application in the fractionation and industrial production of proteins and enzymes with important economic value because of its low cost and easy amplification. Egg white is the cheapest raw material to obtain lysozyme and ovalbumin. Recently, ultrafiltration is often used to separate ovalbumin and lysozyme from egg white.

3. Endotoxin removal
Endotoxin removal is one of the main application forms of ultrafiltration technology in protein purification. The production process of endotoxin is very complex. In the process of practical application, because the medicinal protein produced by prokaryotic expression system is easy to mix with endotoxin produced by bacterial cell wall breaking, and endotoxin, also known as pyrogen, is a kind of lipopolysaccharide. After entering the human body, it can cause fever, microcirculation disturbance, endotoxic shock and other symptoms. In order to protect human health, it is necessary to use ultrafiltration technology comprehensively to remove endotoxins.

Although ultrafiltration technology is widely used in the separation and purification of proteins, it also has certain limitations. If the molecular weight of the two products to be separated is less than 5 times, it cannot be separated by ultrafiltration. If the molecular weight of the product is less than 3kD, it cannot be concentrated by ultrafiltration, because ultrafiltration is usually performed at the minimum molecular weight of the membrane at 1000 NWML.

With the continuous development of bioengineering industry, higher requirements are put forward for downstream separation and purification technology. The traditional methods of vacuum concentration, solvent extraction, dialysis, centrifugation, precipitation and pyrogen removal is not avalible to meet the needs of production any more. Ultrafiltration technology is bound to be more and more widely used because its advantages in protein separation.


Post time: Apr-20-2022
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