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Adenovirus Purification Kit
Adenovirus Purification from any input – cell fraction or media fraction
Rapid purification within 2 to 4.5 hours
No specialized equipment needed (ultracentrifuge not required)
Purify adenovirus cell culture supernatant from 1 mL to 33.5 mL input per prep
Purify adenovirus cell pellet from 1 mL of input per prep
Up to 25X sample concentration
Purification is based on spin column chromatography that uses Norgen’s proprietary resin separation matrix
Adenoviral vectors are useful tools for both in vitro and in vivo gene transfer, and oncolytic viruses based on adenovirus are highly promising for cancer treatment. Norgen’s Adenovirus Purification Kit provides a fast and simple procedure for concentrating and purifying adenoviral vectors from cell lysate and cell culture media. Purification is based on spin column chromatography using Norgen’s proprietary resin as an ion exchanger. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. Adenoviral vectors purified in this manner are highly active for use in transduction experiments.
Norgen’s Adenovirus Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant or a total of 500 mL of supernatant input). Approximately 1 mL of cell pellet can be purified per prep, up to a maximum of 15 mL of cell pellet in total for the entire kit.
Figure 1. HeLa cells were infected with the eluted adenovirus vector expressing the beta galactosidase reporter gene. The cells were fixed and stained 48 hours post infection. The dark staining indicates cellular transduction by the adenoviral vector and expression of the beta galactosidase reporter gene.
Figure 2. Total adenoviral vector eluted for 1, 3, or 15 mL of supernatant containing adenovirus. An increase in titer is observed as the volume of adenovirus containing supernatant increases, demonstrating effective purification at different volumes.
Figure 3. Adenovirus vector was eluted from the kit from various volumes of supernatant: 1 mL, 3 mL, and 15 mL. Three separate elutions (500 µL each) were performed with elution buffer P to elute the adenoviral vector.
At least 3x1011 adenovirus particles as determined by qPCR
At least 3x108 transducing units as determined by transduction assay
Input volume (Supernatant)
1 - 33.5 mL SN per prep (500 mL SN in total)
Input Volume (Cell pellet)
1 mL cell pellet per prep (15 mL in total)
Minimum elution volume
1.3 mL per prep
Time to complete purification
2.5 to 4.5 hours with 1 hour hands on time
In vivo transduction
Storage Conditions and Product Stability
HL-SAN Nuclease should be stored at -20°C upon arrival. Elution Buffer P should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.