Concentration and Purification of Viral Particles

Take a suspension of viral particles mixed with host cell debris.

Centrifuge it to pellet the larger debris. Collect the supernatant, which contains the viral particles along with the smaller debris.

Filter the supernatant to remove the remaining smaller debris.

Add a salt solution containing polyethylene glycol, or PEG, to the filtrate. Invert the tube to mix and incubate.

PEG acts as a molecular crowding agent that pushes the viral particles closer together, promoting their precipitation.

Simultaneously, the salt enhances this effect by neutralizing electrostatic repulsion between the viral particles, enabling them to aggregate.

Centrifuge the tube to pellet the viral aggregates. Discard the PEG-rich supernatant.

Repeat centrifugation, and discard any residual supernatant.

Add a buffer and incubate under agitation to resuspend the pellet.

Aliquot the suspension containing viral particles, and store it at an ultra-low temperature to preserve viral structural integrity and infectivity.

Centrifuge the viral supernatant at 2000 times g for 10 minutes. Afterward purify the viral media by filtering it through a 0.45 micrometer low protein binding syringe filter. Next, add 5X polyethylene glycol