AAV packaging service

We offer AAV packaging services for small-scale experimentation, all the way through to large scale commercial production. Please refer to the list of standard products for more information, or get in touch with our team to discuss custom orders.

The turnaround time for AAV packaging services depends on your specific order details. In general this usually takes 4-6 weeks for the Baculovirus Systema and 6-8 weeks for the Triple Transfection System. To find out more about expected turnaround times before you make an order, please get in contact with our team via our Project Concierge.

Choosing the right AAV Packaging Service will depend on your project requirements. The Triple Transfection system is suitable for smaller volumes of AAV, and can be relatively cost effective. The Baculovirus system is more suitable for larger volumes of AAV, and tends to take less time. For more information on both services see How Does it Work?. You will also need to choose your serotype depending on the tissue type or tropism of your sample. Please refer to the section Choose the Right AAV Serotype which help guide you to choosing an AAV Serotype based on tissue type or tissue tropism. Finally, the best resource you can use is to engage our Project Concierge, who are happily available to help with your specific requirements.

Our AAV packaging services are suitable for a range of different experimentation requirements. Due to its targeted specificity and low immunogenicity the predominant use of AAV technology is in gene therapy applications. AAV can be used in a wide range of pre-clinical and clinical applications in multiple diseases due to its unique biological and biophysical properties. Whether that be in vitro, clinical trials or full-scale production, we can ensure the right products for you. Contact our Project Concierge to discuss your specific project requirements.

Yes. Wild type AAV lacks many proteins necessary to reproduce on its own, making it a prime candidate for gene therapy research. All of Boster’s rAAV’s are replication-defective, non-communicable and can be used in BSL1 and 2 settings. For the Triple Transfection Method we utilise a 3-plasmid co-transfection system which makes the recombinant AAV non-replicable. In the Baculovirus System, we express AAV Rep and Cap genes in trans with our Baculovirus expression system. As Baculovirus is specific to insect cells it poses no threat to mammalian cell lines, and it allows us to make AAV products without the risk of residual adenovirus contamination from helper viruses in triple transfection.

AAV’s major drawback is its very small genome size compared to other existing viral vectors. For instance, compared to adenovirus or lentivirus, AAV is 5 to 10X smaller in genome size. The smaller genome size restricts the size of the gene that can be inserted to less than 4.7 kb in length. To discuss your specific requirements please contact our Project Concierge.

In general, the titer is of more importance because it is a measurement of the number of viral particles per volume.

Purity will be a more important consideration for in vivo studies. If the virus is not pure, it could cause undesired responses to the experimental animals. Our high titer AAVs are suitable for in vivo studies.

For more information on why AAV titer is important click here.

AAVs are known to have low immunogenicity, so it should not be an issue. Compared to other viral vectors (i.e. Lentivirus, Adenovirus), AAV elicits the least amount of immune response in vivo. For more information see Why use AAV?.

Our storage solution for AAVs produced via triple transfection is PBS with 5% Glycerol, which is suitable for in vivo injection. For AAV’s produced using the Baculovirus system storage solution is 1xPBS buffer containing 0.001% pluronic F-68. AAV is stable at 4° C for short-term use and room temperature for immediate use. For long-term storage, AAV should be stored at -80° C for best quality and longevity. Although AAV is a very stable virus, multiple freeze-thaw cycles may reduce the titer of the product.

For Shipping, Ordering and Storage details please see our Sample Collection Guidelines.

If your gene of interest is smaller than 4.7 kilo base pairs (kbp), we can package it into our rAAV products. We do not package gene sequences that may be unsafe to humans or the environment, such as those that are carcinogenic and those that encode for dangerous pathogens. We can always try to package larger AAV vectors as required, however if the GOI exceeds the packaging limit, the titer will be low.

These AAV products are modifications of wild-type AAV5 and AAV8. In each product, the Phospholipase A2 domain encoded by the VP1 Cap gene has been replaced with the corresponding domain from AAV2. Functionally, the activity of this enzyme allows the virus to escape the cellular vesicle once it has been endocytosed into the target cell, therefore avoiding breakdown by lysozymes. AAV2 demonstrates some of the highest phospholipase activity, so we have integrated the AAV2 wild-type domain into our AAV5.2 and AAV8.2 products with no alteration in tropism to increase infectivity

Please request a quote from us. Our controls are produced with a titer of ~2 x 10^13 vg/mL. We can schedule ready-made controls for shipment as soon as we receive a form of payment referencing a quote number. Please contact our Project Concierge for ordering details.

Our production timeline is between 4-8 weeks for custom projects. This timeline will be lengthened if you request a more complex multi-step cloning procedure, or want us to produce your own capsid. For accurate timelines on your specific project please contact our Project Concierge.

We can make AAV to over-express any genes (wildtype, mutant or synthetic) and to silence/knockout any genes from any species if you don’t have any starting material. Our scientists are available to discuss your specific requirement and come up with the recommended AAV constructs for your specific needs. Contact our Project Concierge to discuss your specific requirements.

Initially we will need to know the following in order to send you a quote:

1. The size of the gene you wish to package, and if gene synthesis is required.
2. The AAV serotype you wish to package.
3. The production scale size in vector genome (vg) equivalent to genome copy.
4. If a pre-made control AAV is required, please include the name of the control vector, the AAV serotype, and desired volume and titer.

Please contact our Project Concierge to discuss your specific requirements and we will tailor the service to your needs.

If you accept the quote and wish to proceed with one of Boster’s AAV Packaging Services, we require the following materials:

1. The plasmid with your target gene. 10-20uL of DNA at 0.5ug/uL. We can accept plasmids blotted onto filter paper, however this will require an extra purification step at an extra cost. If you do not have ready-made DNA we can offer a Gene Cloning Service.
2. The text file of the DNA sequence. Files we accept include .dna, .doc, .docx, or any other plain text files. We will keep your information absolutely confidential, and its use will be limited to the project outlined in the quote, unless further requests are made from the original ordering entity.
3. A form of payment.

We use vector genome (vg) as our unit to measure AAV production scale. This unit is similar to genome copy (gc) – it refers to the number of complete copies of the gene of interest that are present in the final suspension.

The titer (in vg/mL), or concentration, of our virus products is typically ~2 x 10^13 vg/mL. However, some genes are either toxic to cells or detrimental to AAV efficiency, and some serotypes (e.g, AAV 2, 3, 6) are low yield. In these cases, the expected titer may be lower.

Both titer and volume can be adjusted to the customer’s requirements, but the price of the product will always depend on the vector genome (vg) amount.

For more details on expected yields for your specific project, contact the Project Concierge.

Our QC details can be found here and are detailed below:

• Titer Test. This is done via qPCR with universal ITR
• Sterility Test.
• Infection QC. If your vector includes GFP and/or RFP under the control of a constitutive promoter (e.g., CMV, PGK, EF1α, etc.), we check whether GFP and/or RFP is expressed after packaging and infection in cells. This additional level of QC is provided free-of-charge.
• SDS Page Gel on AAV proteins VP1, VP2 and VP3 to determine purity
• We can also include an endotoxin assay, and/or have the final product sequence verified at an additional cost.