Adenovirus Packaging

Adenoviral vectors are used for achieving high levels of transgene expression in a wide variety of mammalian cell types, in which the vector remains as episomal DNA without integration into the host genome. High transduction efficiency and high levels of short-term gene expression make adenoviral vectors a preferred tool for in vivo gene delivery and they are often used in gene therapy and vaccination. VectorBuilder has developed a series of proprietary technologies and reagents that have greatly improved adenoviral packaging protocols in terms of titer, purity, viability and consistency, especially for the adenoviral 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 adenoviral vector cloning and packaging needs.
Click here to read detailed introduction of our adenoviral vector systems >>
Click here to view citations of our adenovirus >>
Click here to view user testimonials of our adenovirus >>
How do I obtain adenovirus packaging services?
Click here to view an example of a finished adenoviral vector design >>
Click here to view an example of an adenovirus 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 adenovirus offered

We offer packaging of HAd5 adenovirus and chimeric Ad5/F35 adenovirus.

Price and turnaround

Due to our technological advantage, our adenovirus 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 adenovirus packaging are shown below:

Scale Application Titer & Volume* Price (USD) Turnaround**
Pilot Cell culture >1010 PFU/ml, 10x 25 ul, HBSS buffer $399 20-32 days
Medium Cell culture >1010 PFU/ml, 10x 100 ul, HBSS buffer $599 20-32 days
Large Cell culture >1011 PFU/ml, 10x 100 ul, HBSS buffer $999 20-32 days
Ultra-purified large Cell culture and in vivo >1012 VP/ml, 10x 100 ul, GTS buffer $1499 20-32 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 adenovirus cargo limit (38.7 kb). For sizes above this, the adenoviral genome may be unstable, and rearrangement may occur during packaging. 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.

** Estimated turnaround is the time from production initiation to completion. It does not include waiting time for any customer-supplied materials (e.g. template DNA), QC of such materials, and transit time for shipping final deliverables to the customer.

Deliverables

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

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

If you choose ultra-purified large scale adenovirus packaging (in vivo grade), each order includes the following deliverables:

  • Aliquots of custom adenovirus 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 virus is shown below:
Vector System Control Vector Name Control Vector ID Custom Virus Packaging Scale Corresponding Control Virus Amount Price (USD)
Adenovirus gene expression system pAV[Exp]-CMV>EGFP VB150925-10024 Pilot >1010 PFU/ml, 100 ul in HBSS Free
Medium >1010 PFU/ml, 2x 100 ul in HBSS Free
Large >1010 PFU/ml, 2x 100 ul in HBSS Free
Ultra-purified large >1012 VP/ml, 10x 100 ul in GTS $500
Adenovirus U6-based shRNA knockdown system pAV[shRNA]-EGFP-U6>
Scramble_shRNA
VB161026-1127kyf Pilot >1010 PFU/ml, 100 ul in HBSS Free
Medium >1010 PFU/ml, 2x 100 ul in HBSS Free
Large >1010 PFU/ml, 2x 100 ul in HBSS Free
Ultra-purified large >1012 VP/ml, 10x 100 ul in GTS $500
Adenovirus miR30-based shRNA knockdown system pAV[miR30]-CMV>EGFP:Scramble_miR30-shRNA VB180124-1101mjs Pilot >1010 PFU/ml, 100 ul in HBSS Free
Medium >1010 PFU/ml, 2x 100 ul in HBSS Free
Large >1010 PFU/ml, 2x 100 ul in HBSS Free
Ultra-purified large >1012 VP/ml, 10x 100 ul in GTS $500
Adenovirus CRISPR system pAV[Exp]-CMV>EGFP VB150925-10024 Pilot >1010 PFU/ml, 100 ul in HBSS Free
Medium >1010 PFU/ml, 2x 100 ul in HBSS Free
Large >1010 PFU/ml, 2x 100 ul in HBSS Free
Ultra-purified large >1012 VP/ml, 10x 100 ul in GTS $500
Production, purification and titration

For our adenovirus packaging, the adenoviral vector carrying the gene of interest (GOI) is first linearized by restriction digestion with PacI. The linearized vector DNA is transfected into packaging cells expressing the adenovirus gene E1 to produce recombinant adenovirus, and adenoviral particles are released into tissue culture medium where they are collected. For ultra-purification (in vivo grade), viral particles are further purified and concentrated by cesium chloride (CsCl) gradient ultracentrifugation. We measure the titer using an immunocytochemistry-based approach to detect the adenovirus-specific hexon protein. For ultra-purified adenovirus, we measure the optical density (using OD260) of the viral particles to estimate the titer.

Read more about our titration method >>

Quality control

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

Customer-supplied vectors

If customer-supplied adenoviral 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.