Analysis of the main steps of protein separation and purification
Today, let ’s talk about the pretreatment of the first step of protein separation and purification and cell disruption. The following is introduced by Suzhou protein purification system manufacturers.
When separating and purifying a certain protein, the protein must first be released from the tissues or cells and keep the original natural state without losing its activity. Therefore, appropriate methods should be used to break up tissues and cells. Common methods of breaking tissue cells are:
1. Mechanical crushing method
This method uses mechanical shear to break up the cells. Commonly used equipment include high-speed tissue masher, homogenizer, mortar, etc.
2. Osmotic crushing method
This method swells and breaks cells under hypotonic conditions.
3. Repeated freeze-thaw method
After the biological tissue is frozen, the intracellular fluid freezes and expands, causing the cells to burst. This method is simple and convenient, but it should be noted that those proteins that are sensitive to temperature changes should not use this method.
4. Ultrasonic method
Ultrasonic shaker was used to make the cell membranes unevenly tensioned and the cells were broken.
5. Enzymatic method
Such as destroying microbial cells with lysozyme.
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