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Cell Rupture | Cell Disruption | Cell Lysis Methods

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Deb Shechter
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Sep 12, 2014
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1
min read
Cell Rupture | Cell Disruption | Cell Lysis Methods

Cell Rupture / cell disruption / cell lysis, by any name, rupturing cells in a controlled fashion is essential for both research and manufacturing. Some cells, such as yeasts, are much harder to rupture than others. The method chosen for rupturing the cell also has to take into account the nature of the final product- if the cell is being ruptured in order to obtain an active protein, the method chosen has to be gentle enough to not denature or damage the protein.

Enzymatic digestion is often used in the laboratory as a very gentle method of cell rupture. The cell walls of yeast and bacteria are digested with special enzymes and then the unprotected cell can easily be ruptured by a mild osmotic shock. Enzymatic rupture methods are, in general, too expensive to use in large-scale manufacturing.

Production cell disruption

Chemical methods of rupturing cells are sometimes used. When RNA is the desired product, rupturing cells with a strong chaotropic agent such as urea or guanidine is the approach of choice. These agents denature the proteins, including the many RNA-digesting enzymes present in cells, thus allowing the RNA to be isolated intact. Milder chemical methods, such as using surfactants (e.g. Triton), will allow some active proteins to be isolated. Chemical methods can also be expensive to use in large-scale manufacturing.

Mechanical methods of cell rupture include ball mills, blenders, French presses, homogenizers, and ultrasonic disruption. Ultrasonic disruption is often used in the laboratory with yeast and bacteria, but it is too expensive to use on a larger scale. Blenders and French presses are easy to use in the laboratory, but cannot be readily scaled up for manufacturing. Ball mills are not as popular because the balls need to be removed- an extra step- and usually can only be used once- adding to the cost. Homogenizers are the method of choice for large-scale manufacturing.  

Pion's cell disruption technology comes in both laboratory-sized machines and also in large-scale sizes for manufacturing. The technology is an in-line process that uses cavitation, shear and impact to rupture cells. The forces applied can be precisely adjusted to be as gentle as possible or to be very harsh for more difficult cell types. The sample sizes can vary as well. For example, the Nano DeBEE Laboratory Homogenizer can also handle sample sizes as small as 15 ml.

All aspects of the process can be adjusted- pressure, flow, cavitation, shear, impact and time- allowing the user to fine-tune the method for each application. The precision control of all aspects allows the method to be highly reproducible, and also allows for easy scaling-up. In fact, the Nano DeBEE Laboratory Homogenizer can handle up to 45,000 PSI / 3100 bar for maximum experimentation. The BEE machines are also easy to clean and require little maintenance. 

Don't hesitate to contact us if you have any questions about our cell disruption technology. We have three sizes of machines ideal for any laboratory, and industrial-sized machines for manufacturing. 

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