AAV Packaging

Adeno-associated virus (AAV) is a versatile and popular viral vector tool used for in vitro and in vivo gene delivery. AAVs have emerged as the most effective vehicles for gene therapy due to their ability to transduce a wide variety of mammalian cell types and their low immunogenicity in host organisms. VectorBuilder has developed a series of proprietary technologies and reagents that have greatly improved AAV packaging protocols in terms of titer, purity, viability and consistency, especially for the AAV vector systems used in our vector cloning services. As a result, we have a growing base of highly satisfied customers who come back to us time after time for their AAV vector cloning and packaging needs.
Click here to read detailed introduction of our AAV vector systems >>
Click here to view citations of our AAV >>
Click here to view user testimonials of our AAV >>
How do I obtain AAV packaging services?

Click here to view an example of a finished AAV vector design >>
Click here to view an example of an AAV service proposal >>
Click here to view instructions on designing a vector on VectorBuilder >>
Click here to view instructions on placing an order on VectorBuilder >>

Types of AAV offered

We offer second-generation AAV packaging for the serotypes* listed below. We can package both ssAAV (single-stranded AAV) and scAAV (self-complementary AAV).

Serotypes 1, 2, 3, 4, 5, 6, 6.2, 7, 8, 9, rh10, DJ, DJ/8, PHP.eB, PHP.S, AAV2-retro and AAV2-QuadYF.

* Please note that the ITRs carrying your gene of interest (GOI) is from the AAV2 genome. Different serotypes are distinguished by the capsid protein serotype.

Price and turnaround

Due to our technological advantage, our AAV packaging services have the highest quality in the industry and are also faster and more affordable than those from other suppliers. Prices and turnaround times for different scales of AAV packaging are shown below:

Scale Application Titer & Volume* Price (USD) Turnaround**
Pilot Cell culture >1011 GC/ml, 10x 25 ul, PBS buffer $399 15-25 days
Medium Cell culture >1011 GC/ml, 10x 100 ul, PBS buffer $599 15-25 days
Large Cell culture >1012 GC/ml, 10x 100 ul, PBS buffer $999 15-25 days
Ultra-purified pilot Cell culture and in vivo >1013 GC/ml, 4x 25 ul, PBS buffer $1099 15-25 days
Ultra-purified medium Cell culture and in vivo >1013 GC/ml, 10x 50 ul, PBS buffer $1699 15-25 days

* Our titer guarantee only applies to vectors for which the region being packaged into virus (from 5’ ITR to 3’ ITR) is below the AAV cargo limit (4.7 kb). For sizes above this, ITRs packaged into the virus may be truncated and result in loss-of-function. Additionally, we are not able to guarantee titer for the following vectors:

  • vectors containing sequences that could adversely affect the packaging process such as toxic genes (e.g. proapoptotic genes), genes that compromise the integrity of packaging cells or virus (e.g. membrane proteins that cause cell aggregation), and sequences prone to rearrangements or secondary structures (e.g. repetitive or highly GC-rich sequences);
  • customer-supplied vectors as we have no control over their quality.

** Turnaround time is defined as the time from when an order is accepted to when it is shipped. It does not include waiting time for any materials that the customer needs to supply (e.g. viral vectors) and QC of customer-supplied materials.

Deliverables

If you choose pilot, medium or large scale AAV packaging, each order includes the following deliverables:

  • Aliquots of custom AAV at selected scale
  • Free: aliquots of control virus*

If you choose ultra-purified pilot or ultra-purified medium scale AAV packaging (in vivo grade), each order includes the following deliverables:

  • Aliquots of custom AAV at selected scale
  • "Add-on"purchase (optional): aliquots of ultra-purified control virus*
* The control virus is designed to match the biological application of the custom virus and to be used for testing transduction. For example, if the custom virus overexpresses a gene, then the control virus provided will overexpress EGFP, and if the custom virus expresses an shRNA against a gene, then the control virus provided will express a scramble shRNA. Detailed information for the various control viruses is shown below:
Vector System Control Vector Name Control Vector ID Custom Virus Packaging Scale Corresponding Control Virus Amount Price (USD)
AAV gene expression system pAAV[Exp]-CMV>EGFP VB150925-10026 Pilot >1011 GC/ml, 100 ul in PBS Free
Medium >1011 GC/ml, 2x 100 ul in PBS Free
Large >1011 GC/ml, 2x 100 ul in PBS Free
Ultra-purified pilot >1013 GC/ml, 4x 25 ul in PBS $360
Ultra-purified medium >1013 GC/ml, 10x 50 ul in PBS $560
AAV U6-based shRNA knockdown system pAAV[shRNA]-EGFP-U6>
Scramble_shRNA
VB180117-1020znr Pilot >1011 GC/ml, 100 ul in PBS Free
Medium >1011 GC/ml, 2x 100 ul in PBS Free
Large >1011 GC/ml, 2x 100 ul in PBS Free
Ultra-purified pilot >1013 GC/ml, 4x 25 ul in PBS $360
Ultra-purified medium >1013 GC/ml, 10x 50 ul in PBS $560
AAV miR30-based shRNA knockdown system pAAV[miR30]-CMV>EGFP:Scramble_miR30-shRNA VB180124-1104yzn Pilot >1011 GC/ml, 100 ul in PBS Free
Medium >1011 GC/ml, 2x 100 ul in PBS Free
Large >1011 GC/ml, 2x 100 ul in PBS Free
Ultra-purified pilot >1013 GC/ml, 4x 25 ul in PBS $360
Ultra-purified medium >1013 GC/ml, 10x 50 ul in PBS $560
AAV CRISPR system pAAV[Exp]-CMV>EGFP VB150925-10026 Pilot >1011 GC/ml, 100 ul in PBS Free
Medium >1011 GC/ml, 2x 100 ul in PBS Free
Large >1011 GC/ml, 2x 100 ul in PBS Free
Ultra-purified pilot >1013 GC/ml, 4x 25 ul in PBS $360
Ultra-purified medium >1013 GC/ml, 10x 50 ul in PBS $560
Production, purification and titration

For our AAV packaging, the transfer plasmid carrying the gene of interest (GOI) is co-transfected with our proprietary Rep-cap plasmid and helper plasmid encoding adenovirus genes (E4, E2A and VA) that mediate AAV replication into packaging cells. After a short incubation period, viral particles are harvested from cell lysates and concentrated by PEG precipitation. For ultra-purification (in vivo grade), viral particles are further purified and concentrated by cesium chloride (CsCl) gradient ultracentrifugation. We use a qPCR-based approach to measure AAV titer.
Read more about our titration method >>

Quality control

For each AAV produced by VectorBuilder, quality control includes titer measurement, sterility testing for bacteria and fungi, and mycoplasma detection. If the transfer vector encodes a fluorescent protein, we would transduce cells with virus and detect corresponding fluorescence. Additionally, for ultra-purified AAV, we routinely check virus quality by SDS-PAGE analysis and endotoxin assay.

Customer-supplied vectors

If customer-supplied AAV vectors are needed, please send us the vectors following the Materials Submission Guidelines. Please strictly follow our guidelines to set up shipment to avoid any delay or damage of materials. All customer-supplied materials undergo mandatory QC by VectorBuilder which may incur a $100 surcharge for each item. Please note that production may not be initiated until customer-supplied materials pass QC.

Questions

Please refer to our Vector FAQ for commonly asked technical questions. To get additional information about what we can offer, please contact VectorBuilder's service team following the contact information here.