How To Freeze Dry RNA?
How to freeze dry RNA — background
The application of RNA is a representative of modern molecular medicine, and it also shows some superior properties relative to the application of DNA. It is generally known that the transfection of DNA molecules can cause serious complications. However, if RNA, especially mRNA, is used instead of DNA, these complications will not occur. However, the main disadvantage of using RNA is its instability. RNA is easily hydrolyzed by ubiquitous ribonuclease or divalent cations and is usually rapidly degraded. To avoid this rapid degradation, RNA (in solution) is usually stored at -20°C or even -80°C and ribonuclease-free conditions. However, such storage conditions are costly and not sustainable. Another method for stabilizing RNA is the freeze-drying of RNA. Lyophilization is a known and recognized method worldwide, and it can be used to enhance the storage stability of temperature-sensitive biomolecules. How to freeze dry RNA? Here is the process.
How to freeze dry RNA — the freeze-drying process
a) Provide a liquid containing at least one RNA and at least one lyoprotectant;
b) Load the provided liquid into the chamber of a lyophilization machine;
c) Cooling the liquid to freezing temperature, where the cooling is performed at a certain rate;
d) Freeze the liquid to obtain a frozen product;
e) Reduce the pressure in the freeze-drying chamber to a pressure lower than atmospheric pressure;
f) Drying the frozen liquid obtained in step d to obtain a lyophilized composition containing at least one RNA and one lyoprotectant;
g) Equilibrate the pressure in the freeze-drying chamber with atmospheric pressure, and remove the freeze-dried composition obtained in step from the chamber of a freeze dryer lyophilizer.
How to freeze dry RNA — freeze-drying notes
Before step g, the freeze-drying chamber needs to be filled with inert gas. These gases can be nitrogen, carbon dioxide, helium, neon, argon, xenon, and krypton.
In step e, the pressure in the freeze-drying chamber needs to be reduced to the primary drying pressure, and this pressure value needs to be maintained during the primary drying process. Subsequently, the pressure in the freeze-drying chamber is reduced to the secondary drying pressure, and the pressure needs to be maintained during this period to ensure that the activity of RNA is not affected.
In step f, the heating rate has an effect on the quality of the lyophilized product. Therefore, we must choose an appropriate heating rate to ensure the integrity and biological activity of RNA. Generally speaking, the heating rate is best controlled at 5°C/hour to 10°C/hour.
How to freeze dry RNA — features of freeze-dried RNA
Excellent integrity of RNA.
Stable chemical properties, suitable for storage and environments without any cooling equipment.
Suitable for use on an industrial scale. The RNA composition can be advantageously stored, transported, and used.
How to freeze dry RNA, Hawach has given the answer. As a freeze dryer manufacturer, Hawach is willing to share all the knowledge they know to the dear customer. If you have any suggestions, remember to contact us!