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Virus Purification Kits

Virapur, LLC (San Diego, CA, USA) is a virus production and purification company. Their expert virologists have years of experience in producing and purifying adenovirus and many other human and animal viruses.

Features

Short time for purification
Unnecessary CsCl gradients
Easy operation

Adenovirus Purification Kit


Figure 1	Figure 2

1.	HEK293 cells or their variants should be seeded into tissue culture flask at approximately 4 x 10⁴ cells per cm². The cell monolayer will become nearly confluent within approximately 2 to 4 days. When the monolayer is 95 to 100% confluent, remove most of the media in the flask or vessel and replace with new growth media with serum reduced to 2%.
2.	Add your adenovirus (3 PFU per cell) to the culture to initiate the infection. Incubate for 3 to 5 days until cytopathic effect is complete.
3.	Pool all the cell lysate and media into one capped vessel and freeze and thaw at least two times. Pour the entire cell lysate into a centrifuge tube or bottle. Spin at 2500 to 2800 rpm for 30 minutes. Collect the supernatant into another clean bottle.
4.	Add 1000 Kunitz Units of DNase I to 100 ml of the clarified crude virus solution. Gently invert the solution and incubate at 37^oC for 30 minutes.
5.	Carefully place the provided single glass fiber pre-filter on top of the membrane in the top of the filter unit. Attach the bottle top filter to a vacuum source and pre-wet the filters with approximately 25 ml of PBS or media.
6.	Carefully pour the spun virus supernatant into the filter unit to clarify infected cell lysate.

7.	Pour the filtered supernatant into a new clean bottle and measure the volume. Add an equal number of ml of Dilution Buffer A to the filtered supernatant. Mix gently but thoroughly.
8.	Wet the interior of the purification filter by passing about 5 ml of sterile PBS (user provided) through the filter with a syringe. Attach the purification filter to the tubing and syringe. Pass the diluted virus supernatant through the purification filter at a rate of about 20 ml per minute. (Figure 1) Pull the Wash Buffer B through the purification filter at a rate of about 20 ml per minute.
9.	Draw at least 1.5 ml of Elution Buffer C into 3-5 ml syringe. Attach the syringe to the filter unit. Position a tube under the purification filter to catch the elution buffer containing virus. (Figure 2)
10.	Slowly pass the elution buffer through the filter. The final 0.5 ml can be drawn up and down very slightly before being expelled from the filter to release all virus on the filter.
See instruction for more details.
Product instruction

Adenovirus Mini Purification Kit

Figure 3

Product instruction

AAV Purification Kit


Diluted virus supernatant
Figure 4	Figure 5

1.	HEK293 cells or their variants can be grown in tissue culture flask at approximately 4 x 10⁴ cells per cm². The cell monolayer will become nearly confluent within approximately 2 to 4 days. You will transfect the cultures according to your own protocol with multiple plasmids.
2.	After transfection, harvest the cultures within 2 to 5 days. At harvest, pool all the cell lysate and media into one capped vessel and freeze and thaw at least three times. After the third thaw, pour the entire cell lysate into a centrifuge tube or bottle. Spin at 2500 to 2800 rpm for 30 minutes. Collect the supernatant into another clean bottle.
3.	Remove much of the contaminating DNA by adding 1000 Kunitz Units of DNase II to 100 ml of the unpurified virus solution. Gently invert the solution and incubate at 37^oC for 30 minutes.
4.	Carefully place the provided single glass fiber pre-filter on top of the membrane in the top of the filter unit. Attach the bottle top filter to a vacuum source and pre-wet the filters with approximately 25 ml of PBS or media.
5.	Carefully pour the spun virus supernatant into the filter unit to clarify infected cell lysate.
6.	Measure the volume of the filtered supernatant. Add the calculated volume of Dilution Buffer 1 to the filtered supernatant. (Dilution Buffer 1: the filtered supernatant =1:9) Mix gently but thoroughly.

7.	Attach the Large/Small purification filter assembly to the tubing and syringe. and wetting it with PBS. Pass the diluted virus supernatant through the purification filter at a rate of about 20 ml per minute (Figure 4). Disconnect the large filter, and pass the wash buffer through to clean the SMALL filter at a rate of about 20 ml per minute.
8.	Obtain two new 3-5 ml syringes. Draw at least 1.5 ml of Elution Buffer 3 into one syringe. Attach the two syringes to the SMALL filter using the female luer adaptor provided in the plastic bag to attach the second syringe to the SMALL filter. (Figure 5)
9.	Pass the elution buffer slowly from the first syringe through the filter into the second syringe. Slowly pass the elution buffer back to the first syringe. Blow a little air through the filter to collect all elution into one syringe.
See instruction for more details.

Product instruction

Ordering information

Adenovirus Purification Kit

Product name	Storage	Product number	PKG Size
Adenovirus Purification Kit, Single-Use	RT	003051	1kit
Adenovirus Purification Kit, Four-Use	RT	003054	1kit
Adenovirus Large Scale Purification Kit, Single-Use	RT	003056	1kit
Adenovirus Mini Purification Kit, Four-Use	RT	003058	1kit
Adenovirus Mini Purification Kit, 24-Use	RT	003059	1kit

AAV Purification Kit

Product name	Storage	Product number	PKG Size
AAV Purification Kit, Single-Use	RT	003061	1kit
AAV Purification Kit, Three-Use	RT	003063	1kit

Printed Version

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