AAV Purification Kits

Cat. 66100, 63200, 63300, 63250

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AAV Purification Kits

Cat. 66100, 63200, 63300, 63250

For concentrating and purifying AAV vectors from cell lysate and cell culture media

AAV Purification from any input – cell fraction or media fraction
High AAV recovery, up to 90%
No specialized equipment needed
Purification from a variety of AAV serotypes (including AAV6 and AAV9)
Yields highly active AAV for in vivo and in vitro experiments
Purification is based on spin column chromatography that uses Norgen’s resin separation matrix

For research use only and NOT intended for in vitro diagnostics.

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AAV Purification Kit
(Cat. 66100) 15 preps

AAV Purification Mini Kit
(Cat. 63200) 20 preps

AAV Purification Midi Kit
(Cat. 63300) 4-8 preps

AAV Purification Maxi Slurry Kit
(Cat. 63250) 1-10 preps

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Product Format
Norgen Biotek offers kits in different standardized formats. You can view and compare kit components in the Components Table in the Product Overview section. For more information on format types, please visit our Format Information page.

Product Size

Kit Specifications

You have selected: Cat. 66100

Kit Specifications
Resin Binding Capacity (total per kit)	At least 5 x 10¹⁰ AAV particles as determined by qPCR
AAV Vector Serotype	AAV6, AAV9 and others
Input Type	Cells, media
Input Volume (AAV supernatant)	1 - 33.5 mL SN per prep (500 mL SN in total)
Input Volume (AAV cell pellet)	1 mL cell pellet per prep (15 mL in total)
Time to Complete Purifications	2.5 to 4.5 hours with 1 hour hands on time
In vivo transduction	Yes

Storage Conditions and Product Stability

HL-SAN Nuclease should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 2 years after the date of shipment.

You have selected: Cat. 63200

Kit Specifications
Column Binding Capacity	At least 5 x 10⁹ AAV particles as determined by qPCR
AAV Vector Serotype	Any
Average Recovery	> 80%
Input Type	Cells, media, or mixed
Input Volume	0.5 mL - 8 mL
Minimum Elution Volume	200 µL
Time to Complete Purifications	1 - 2 hours

Storage Conditions and Product Stability
DNAse I and RNAse A should be stored at -20°C upon arrival. Elution Buffer O should be stored tightly capped at 4°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. Once opened, the solutions should be stored at 4°C. This kit is stable for 1 year after the date of shipment.

You have selected: Cat. 63300

Kit Specifications
Column Binding Capacity	At least 1 x 10¹⁰ AAV particles as determined by qPCR
AAV Vector Serotype	Any
Average Recovery	> 80%
Input Type	Cells, media, or mixed
Input Volume	8 mL - 45 mL
Minimum Elution Volume	1 mL
Time to Complete 4 Purifications	2 - 2.5 hours

You have selected: Cat. 63250

Kit Specifications
Column Binding Capacity	At least 1 x 10¹⁰ AAV particles as determined by qPCR
AAV Vector Serotype	Any
Average Recovery	50 - 80%
Input Type	Cells, media, or mixed
Input Volume	90 mL - 900 mL
Minimum Elution Volume	1 mL - 10 mL
Time to Complete Purifications	2.5 - 3.5 hours

Supporting Data

Figure 1.In vitro transduction. HTX cells transduced with 50 µL eluted vector from the Norgen AAV Purification Kit after purification of cell culture supernatant containing AAV. Both AAV9 and a bovine AAV capsid (isolate AAV-Ca) were tested in vitro on HTX cells. The vector encoded an alkaline phosphatase reporter gene driven by the CAG promoter. Dark/purple staining represents cells that have been transduced by AAV bearing the alkaline phosphatase reporter gene.

Figure 2. In vivo testing of AAV vector purified using the Norgen AAV Purification Kit in the mouse lung. 2 x 1011 ss genomes of an AAV9 vector encoding an alkaline phosphatase reporter gene was delivered via the intranasal route. 1 month post transduction, mouse tissues were harvested. Areas of transduction in the mouse lung are shown by the dark/purple staining where the alkaline phosphatase reporter gene was being expressed.

Figure 3. Transduction of HTX cells using Norgens AAV Purification Mini Kit. Microscopic view of HTX cells transduced with biologically active AAV vector after purification using Norgen's AAV Purification Mini Kit (dark purple represents alkaline phosphatase transgene expression).

Figure 4. Linear Scaling of Norgen's AAV Purification Mini Kit From 2.5 mL to 15 mL Input. Purification using Norgen's AAV Purification Mini Kit across different input volumes consisting of mixed cells and supernatant ranging from 2.5 mL to 15 mL. Increased input volumes resulted in no loss of vector as demonstrated by linear scaling across the different volumes.

Figure 5. Transduction of HTX cells with Norgen's AAV Purification Mini Kit with different input volumes of mixed cells and supernatant (0.5 mL. 2.5 mL, 10 mL and 15 mL). Microscopic view of HTX cells transduced with biologically active AAV vector after purification using Norgen's AAV Purification Mini Kit (dark purple represents alkaline phosphatase transgene expression).

Figure 6. Purification from Both Cell Pellet and Supernatant. Purification of AAV vector from both Cell Pellet and Cell Media/Supernatant fraction.

Figure 7. Linear Scaling of Norgen's AAV Purification Midi Kit From 2 mL to 50 mL Input. Purification using Norgen's AAV Purification Midi Kit across different input volumes of mixed cells and supernatant ranging from 2 mL to 50 mL. Increased input volumes resulted in no loss of vector as demonstrated by linear scaling across the different volumes.

Figure 8. Transduction of HTX cells with Norgen's AAV Purification Midi Kit with different input volumes (2 mL, 10 mL and 50 mL) of mixed cells and supernatant. Microscopic view of HTX cells transduced with biologically active AAV vector after purification using Norgen's AAV Purification Midi Kit (dark purple represents alkaline phosphatase transgene expression).

Figure 9. Cellular Transduction of HTX cells with 50 µL of AAV purified using Norgens AAV Purification Maxi Slurry Kit. Dark purple staining indicates expression of a human placental alkaline phosphatase reporter gene from AAV transduced cells.

Figure 10. Supernatant from cells transfected with AAV production plasmids was purified using the Norgen AAV Purification Maxi Slurry Kit. Three different volumes were tested: 40, 60 and 80 mL, demonstrating scalable purification of AAV vector over increasing volumes.

Click for expanded view

Protocols:

(66100) AAV Purification Kit - Protocol (15 preps)

(63200) AAV Purification Mini Kit - Protocol (20 prep)

(63300) AAV Purification Midi Kit - Protocol (4-8 prep)

(63250) AAV Purification Maxi Slurry Kit - Protocol (1-10 preps)

Application Notes:

Purification of Adeno-associated Virus (AAV) Vectors Using Norgen’s AAV Purification Kit

Purification of Adeno-associated Virus (AAV) Vectors Using Norgen’s AAV Mini and Midi Purification Kits

Product Overview
Documentation

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AAV Purification Kits

Recombinant adeno-associated virus (AAV) vectors are highly promising tools for both in vitro and in vivo gene transfer. Norgen’s AAV Purification Kits provide fast and simple procedures for concentrating and purifying AAV vectors from cell lysate and cell culture media. Purification is based on precipitation onto Norgen Biotek’s proprietary resin. Contaminating cellular debris is largely removed from the sample via a centrifugation step, while contaminating DNA and RNA is reduced using enzymatic digestion. AAV vector purified in this manner is highly active for use in in vitro and in vivo transduction experiments.

AAV Purification Kit

Norgen’s AAV Purification Kit contains sufficient materials for 15 preparations (33.5 mL per prep of supernatant (SN) 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. Up to 33X sample concentration.

AAV Purification Mini Kit

Each spin column is able to concentrate and purify AAV from 0.5-8 mL of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (200 µL). Preparation time for 4 samples is 1.5 hours, with 45 minutes of hands-on time.

AAV Purification Midi Kit

Each spin column is able to concentrate and purify AAV from 8 mL up to 45 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 50X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1 mL). The kit may be used to purify up to 8 x 25 mL or 4 x 45 mL of samples using the included columns. Preparation time for 4 samples is approximately 2 to 2.5 hours, with 1.5 hours of hands on time.

AAV Purification Maxi Kit (Slurry Format)

Each spin column is able to concentrate and purify AAV from 45 mL to 90 mL of input consisting of cell pellet, cell culture media, or cells and culture media mixed together. Up to 200X sample concentration. AAV vector purified in this manner is highly active for use in in vitro transduction experiments, and is eluted into a small volume (1-10 mL) using the optional concentration step. The kit may be used to purify up to 1 x 900 mL samples or 10 x 45-90 mL samples using the included columns. Preparation time for 1 x 900 mL sample is approximately 2.5 to 3.5 hours, with an optional concentration step requiring an additional 30 min.