アデノウイルスパッケージング

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, often used in gene therapy and vaccination. VectorBuilder has developed a series of proprietary technologies and reagents that 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 adenovirus packaging from E1/E3 deleted HAd5 adenoviral vector.

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 time for different scales of adenovirus packaging are shown below:

スケール 推奨の使用系 タイター & 容量* 価格(税抜き) 作業日数**
パイロット 培養細胞 >1010 PFU/ml, 10x 25 ul, HBSS buffer 64,000円 20-32 日
中容量 培養細胞 >1010 PFU/ml, 10x 100 ul, HBSS buffer 89,000円 20-32 日
大容量 培養細胞 >1011 PFU/ml, 10x 100 ul, HBSS buffer 139,000円 20-32 日
超純粋大容量 培養細胞とin vivo >1012 VP/ml, 10x 100 ul, GTS buffer 201,000円 20-32 日

* Our titer guarantee only applies to vectors for which the region being packaged into virus (from 5’ ITR to 3’ ITR) is below adenovirus cargo limit (38.7 kb). For sizes over this range, 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.

** 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).

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 of the 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 TBD
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 TBD
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 TBD
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 TBD
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 to tissue culture medium and collected from there. 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 test for bacteria and fungi, and mycoplasma detection. If the adenoviral vector encodes a fluorescent protein, we would transduce cells with the for detection of the corresponding fluorescence. Additionally, for ultra-purified adenovirus, we routinely check virus quality by 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 12,500JPY surcharge per material. Please note that production may not be initiated until customer-supplied materials are received and pass QC.

Questions

Please refer to our Vector FAQ for commonly asked technical questions. To get additional information on our offerings, please contact VectorBuilder service team following the contact information here.