Hello
I am trying to extract a virus from some wastewater. Is it possible to use PEG and NaCl at lower speeds? Or are there any alternatives that aren't costly? I don't have access to a high speed centrifuge.
Thank You
Principle:
PEG is well known as good inductor of attractive interactions that crystallize virus in the interpolymer spaces between PEG molecules. Also, When the NaCl concentration is increased, some of the water molecules are attracted by the salt ions, which decreases the number of water molecules available to interact with the charged part of the protein. As a result of the increased demand for solvent molecules, the protein–protein interactions are stronger than the solvent-solute interactions; the protein molecules coagulate by forming hydrophobic interactions with each other. This process is known as salting out.
TBS, or TES, or PBS, buffer is used for eliminating the PEG polymer remnants and also, it is employed to keep the pH within an optimal range to maintain the native structure of the virus .
Methods:
To the 100-mL primary concentrate, 14 g PEG 8000 and 1.17 g NaCl (for a final sample concentration of 0.2 M NaCl) is added and shaken vigorously by hand (~ 5 min, room temperature) until dissolved. Samples are then shaken overnight (16–18 h, 200 RPM, 4 °C), centrifuged (30 min, 4500×g, 4 °C), and the supernatant carefully removed. The pellets were resuspended in 10–20 mL PBS, pH 7.4 and assayed.
Virus Precipitation using Kit:
PEG Virus Precipitation Kit can be used for small lab samples or large scale virus preparation with high yield and high viral titer.
A quick, easy and inexpensive method is desired to concentrate virus and remove impurities • Easy, convenient and time-saving method to concentrate virus without ultra-centrifugation • Virus can be concentrated over 100 folds • The whole process uses non-toxic reagents
Thank you Santhana Krishnan . This is a great answer! However, my lab space is very limited and do not have a machine that shakes samples overnight. Is this step crucial or is there an alternative to that? Thank you again.
@Antonio Zapata . Sir, try to consider stirring with a bar magnet at low rpm.
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