Purification of Virus from the Allantoic Fluid of Embryonated Chicken Eggs

Begin with a reservoir containing allantoic fluid harvested from virus-infected chicken eggs.

This fluid contains virus particles along with host-derived contaminants.

Pass the fluid through a depth filter to capture large contaminants while allowing smaller contaminants and viruses to pass through.

Then, load the filtered fluid into a filtration system, where it flows tangentially across a membrane surface.

Smaller contaminants pass through the membrane, while the virus is retained and concentrated.

Load the concentrated virus onto a density gradient and centrifuge it. The virus forms a distinct band where its density matches the gradient.

Collect the virus-rich band in a dialysis cassette and place it in a buffer. The gradient components diffuse out, while water from the buffer diffuses in through the semipermeable membrane.

Next, place the cassette in a hypertonic solution to draw out water and concentrate the purified virus.

For the purification of Newcastle Disease Virus or NDV, first, prime the previously set experimental system by running 50 milliliters of PBS through it.

Stop the pump when approximately five milliliters of PBS are left in the tube. Next, attach a depth filter with a