Viral vector platform
Adenovirus packaging
Adenoviral vectors deliver large transgenes without integrating into the host genome, which suits short-term high-level expression and any workflow where genome integration is undesirable. FuBio packages Ad5-based vectors carrying reporter, gene-editing and control cassettes.
What Are Adenoviruses?
Adenoviruses are non-enveloped viruses with an icosahedral capsid of roughly 90 to 100 nm, carrying a linear double-stranded DNA genome of about 36 kilobases with a terminal protein covalently attached at each end. Human adenovirus type 5 (Ad5) is the best characterized serotype and the backbone used for most adenoviral vectors.
Adenoviral vectors are engineered to be replication-deficient. In first-generation Ad5 vectors the E1 region required for replication is deleted and replaced by the expression cassette, and E3 is deleted to create packaging room, so the vector can only replicate in complementing producer cells such as HEK293 that supply E1 in trans. The genome stays episomal: expression is strong and rapid, does not depend on cell division, and is not passed on through integration.
Adenoviral Vector Packaging and Production
FuBio builds adenoviral vectors on a replication-deficient human adenovirus type 5 backbone. The expression cassette is introduced on a shuttle plasmid and combined with the E1-deleted adenoviral genome before the vector is rescued in producer cells:
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Shuttle Plasmid
Carries the expression cassette — reporter, transgene or CRISPR nuclease — flanked by the adenoviral inverted terminal repeats and the packaging signal, with the E1 region replaced by the cassette.
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Adenoviral Backbone Plasmid
Supplies the remainder of the Ad5 genome with E1 and E3 deleted. Recombination with the shuttle plasmid reconstitutes the full vector genome, and the construct is linearised before transfection so that viral rescue can start.
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Producer Cells (HEK293 / 293A)
Complement the deleted E1 functions in trans, so the vector genome is replicated and packaged into capsids while the vector itself remains replication-deficient in ordinary target cells.
After transfection, the vector is amplified through successive passages in producer cells, then released by lysis, clarified and purified by caesium chloride gradient ultracentrifugation or chromatography. Because the capsid is exceptionally stable, adenovirus tolerates purification and concentration far better than enveloped vectors.
Infectious titer is reported in PFU/mL, with particle concentration available as VP/mL, and every lot is tested for sterility, mycoplasma, endotoxin and replication-competent adenovirus.
Adenovirus Packaging Service Advantages
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Up to 1E+11PFU/mL
Product titers of up to 1E+11 PFU/ml, with particle concentration and infectious titer reported per batch and PFU-based potency where applicable.
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100+Vector Options
Access to over 100 vectors featuring diverse regulatory and packaging systems, including Cas9 expression vectors alongside EGFP, GFP, tdTomato, mCherry and luciferase control constructs.
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LargePayload Capacity
Streamlined processes for the rapid release of Ad5 vectors suited to transient expression in dividing and non-dividing cells, including cassettes that exceed the packaging limit of integrating vectors.
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QualityControl
Ten comprehensive quality control measures and six rigorous testing protocols to ensure the highest standards.
Applications of Adenoviral Vectors
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1
High-Level Transient Expression
Driving strong, rapid expression in dividing and non-dividing cells for short-term assays where integration is undesirable.
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2
CRISPR-Cas9 Delivery
Delivering nucleases and guide cassettes, including constructs too large for integrating vectors.
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3
Reporter and Control Vectors
Fluorescent and luciferase control constructs that mark infection efficiency and normalize delivery across experiments.
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4
Vaccine and Immunotherapy Research
Immunogenic, non-integrating delivery suited to antigen expression, oncolytic designs and immune-activation studies.
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5
In Vivo Delivery to Liver, Lung and Tumor
Systemic and local delivery in animal models, with capsid choice matched to receptor profile and target tissue.
Adenoviral Vector Features
| Feature | Adenovirus |
|---|---|
| Gene Expression | Strong and rapid, transient (non-integrating) |
| Immunogenicity | High |
| Payload Capacity | Large – beyond integrating-vector limits |
| Tropism (Targeting ability) | Broad – Ad5 and chimeric capsids such as Ad5/F35 |
| Best used for | Short-term high-level expression, large cassettes |
Production and quality control
- Vectors are amplified in HEK293 or HEK293A producer cells, lysed, clarified and purified by CsCl gradient ultracentrifugation or chromatography.
- Infectious titer is reported in PFU/mL; particle concentration (VP/mL) and the particle-to-PFU ratio are available on request.
- Every lot is tested for sterility, mycoplasma and endotoxin, and screened for replication-competent adenovirus (RCA) — the critical safety attribute for first-generation Ad5.
- Because the capsid is exceptionally stable, adenovirus tolerates purification and concentration far better than enveloped vectors; titers of 1E+10 PFU/mL or higher are routine.
Our Service Options
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Standalone Adenoviral Packaging
For researchers with existing adenoviral transfer vectors, we provide adenovirus packaging services using customer-supplied constructs, subject to compatibility review.
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Vector Construction & Adenoviral Packaging
For projects requiring additional support, our integrated service covers vector construction and adenovirus packaging, offering a streamlined solution from vector design requirements to adenoviral vector production.
Service Specifications
| Specification | Information |
|---|---|
| Vector Type | Replication-deficient human adenovirus type 5 (Ad5), E1 and E3 deleted |
| Producer Cells | HEK293 / HEK293A |
| Catalog Vectors | 14 vectors available from stock |
| Vector Titer | 1E+10 PFU/mL or 1E+13 GC/mL, per construct |
| Volume | 200 μL |
| Quality Control | Infectious titer (PFU/mL) and particle concentration (VP/mL); sterility, mycoplasma, endotoxin and RCA screening |
| Turnaround Time | Quoted per project in the statement of work |
| Application | In vitro / In vivo Research |
Start Your Adenovirus Packaging Project
Whether you need packaging for an existing transfer vector or a comprehensive vector construction and packaging solution, our team is ready to help you identify the right approach for your research.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
Experimental validation
Adenoviral performance is confirmed in vivo and at the cell level. The images below show biodistribution five days after tail-vein injection of Ad5-WT and hepatotoxicity-reducing Ad5 mutants in Balb/c mice, and flow cytometric analysis of Ad5/F35 chimeric vector infection in Jurkat cells.
Resources
User instructions
A condensed bench protocol for infecting cultured mammalian cells with FuBio Ad5 preparations. The full version ships with each order and the current PDF can be requested through the quote form.
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1
Prepare the cells
Seed cells to reach 70 to 90 percent confluence at infection — adenovirus tolerates higher confluence than most vectors. Verify your line expresses CAR if transduction efficiency is unexpectedly low; many primary and hematopoietic cells do not.
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2
Thaw the stock
Thaw on ice or in the hand, mix gently and keep cold. Avoid freeze-thaw cycles: each one reduces infectious titer measurably even though the particle count stays constant.
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3
Infect
Remove most of the medium and add virus diluted in a small volume of serum-free or complete medium so the inoculum just covers the monolayer. A starting MOI of 1 to 10 PFU per cell suits easy lines; 10 to 100 may be needed for refractory ones. Rock gently for 1 to 2 hours at 37 degrees, then add complete medium to working volume.
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4
Harvest the readout
Expression peaks 24 to 72 hours after infection depending on the promoter and the assay. Collect cells or supernatant inside that window; expression then decays as the episomal genomes are diluted and the innate immune response builds.
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5
Dispose correctly
Inactivate waste with hypochlorite or a validated chemical disinfectant before it leaves the cabinet; non-enveloped capsids survive detergent steps that inactivate enveloped viruses.
Bench guidance only: adenovirus is highly immunogenic in vivo and CAR-dependent in vitro — verify permissiveness of your target cells, and work under the biosafety conditions approved by your institution.
Why should I choose adenoviral vectors for my study?
Adenoviral vectors infect dividing and non-dividing cells with high efficiency, carry larger cassettes than integrating vectors, and can be produced at very high titer. Expression starts within hours rather than days and is strong, because the genome stays episomal and is transcribed at high levels. Choose adenovirus when you need short-term, high-level expression and do not want genome integration; choose AAV or lentivirus when you need expression to persist for weeks in vivo or through many passages.
What are adenoviral vectors used for?
Typical uses include transient high-level transgene expression, delivery of CRISPR-Cas9 components and other large cassettes, reporter and control constructs for infection-normalized experiments, and vaccine or immunotherapy research where the strong innate response to the capsid is an advantage rather than a problem. They are also used for in vivo delivery to liver, lung and solid tumor after local or systemic administration.
What is the difference between PFU/mL and VP/mL?
PFU per milliliter is the infectious titer, measured by counting plaques or infectious foci on a permissive cell layer, and it is the number to dose from. VP per milliliter is the total particle count, which is always higher; the ratio between the two tells you how much of the preparation is infectious and is a useful indicator of how a lot has been handled. We report infectious titer per batch and can provide particle concentration and the particle-to-PFU ratio on request.
Why does adenovirus infect some of my cells poorly?
Ad5 entry depends primarily on the coxsackievirus and adenovirus receptor, CAR, so cells with low CAR expression — many hematopoietic and primary cells — are poorly transduced by Ad5. Chimeric capsids such as Ad5/F35, which use CD46 instead, are the usual fix for these targets. Before assuming failure, check the receptor profile of your line, raise the multiplicity of infection, and confirm the cells are at the confluence that suits the assay.
What is the biosafety profile of adenoviral vectors?
First-generation Ad5 vectors are replication-deficient outside E1-complementing cells, and every lot is screened for replication-competent adenovirus. They are nonetheless commonly handled at biosafety level 2 or the equivalent approved by your institution, and vectors expressing hazardous transgenes may require additional containment. The capsid is non-enveloped and resists detergents that inactivate enveloped viruses, so waste should be inactivated with hypochlorite or another validated disinfectant. All products are for research use only.
How should adenoviral vectors be stored and handled?
Store single-use aliquots at minus 80 degrees Celsius and avoid freeze-thaw cycles, which lower infectious titer even though the particle count stays constant. Thaw on ice, mix gently rather than vortexing, and keep the aliquot cold while you prepare dilutions in serum-free or complete medium. Because the capsid is stable, short handling at room temperature is tolerated better than for enveloped vectors, but the virus should still go back onto ice promptly.
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beta-Arrestin 2 mediates arginine vasopressin-induced IL-6 induction via the ERK1/2-NF-kappaB signal pathway in murine hearts
Acta Pharmacologica Sinica
doi: 10.1038/s41401-019-0292-y
School of Pharmacy, Nantong University
FuBio supplied adenoviral vectors for in vitro and in vivo pathway manipulation.
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The YAP/HIF-1alpha/miR-182/EGR2 axis is implicated in asthma severity through the control of Th17 cell differentiation
Cell & Bioscience
doi: 10.1186/s13578-021-00560-1
Shanghai Children\u2019s Medical Center, Shanghai Jiao Tong University School of Medicine
FuBio supplied the adenoviral vectors used to modulate the axis in Th17 differentiation assays.
Feedback from researchers who have used our adenovirus packaging service will be published here.
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