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Custom Adenovirus Production Introduction Recombinant adenoviruses (rADVs) have been widely used by researchers in the areas of biological research. Among several adenovirus serotypes, human adenovirus 5 has been well characterized and become a popular gene delivery tool. The recombinant human adenovirus 5 particles are usually E1/E3 deletion to make them replication deficient. Sequence of interest can be inserted into either E1 or E3 region. rADV has a larger packaging capacity compared with recombinant AAV or lentivirus. It can almost infect any mammalian cell and is good for both in vitro cell experiments and in vivo animal tests. However, unlike recombinant lentivirus, rADV does not integrate into cell genome. Thus it can only be used for transient expression. RGBiotech has a team of experts working on adenoviuses for years. We can help you to get the optimal adenovirus particles for your special project. Either small scale or pilot scale adenovirus production is available at our company. RGBiotech also offers Oncolytic Adenovirus (CRAds, Conditionally-Replicative Adenoviruses) production. Don’t hesitate to contact us if you have any requirement on adenovirus. Features - One-stop adenovirus service: start from any step of adenovirus production- Adenovirus backbone: human adenovirus 5 - Choice of adenovirus shuttle plasmid construction: ->E1 region: wild type or deleted with MCS ->E3 region: wild type or deleted with MCS ->E4 region: wild type or deleted with MCS - Provide multiple options of promoters: promoterless, CMV, CAG, Synapsin, EF1α, UBC, ALB1.4, ApoE/AAT1, CaMKII, ELA1, Enh358MCK, cTNT, GFAP, MBP, SST, TBG, αMHC, hRPE, mIP1, tMCK, H1, U6 - Choice of adenovirus fibers: wild type Ad5 fiber, hybrid Ad5/3 fiber, hybrid Ad5/35 fiber - Insert type: mammalian genes (human/mouse/rat etc.), virus gene (e.g., HBV genome), shRNA/miRNA encoding sequence etc. - Large insert capacity: up to ~ 11.2 kb - High titer: up to 1 x 10^11 PFU/mL, or 1 x 10^15 VP/mL - High purification via CsCl / Iodixanol ultra-centrifugation gradient, or by chromatography - Suitable for both in vivo animal injection tests and in vitro cell infection experiments - Fast turnaround time: typical 3~ 5 weeks - Save time from investigation of adenovirus marketing and provide reasonable price Service Workflow - Communication with customers to discuss the fit-to-project solution- Complete adenovirus production outline: -> sequence design and synthesis -> sequence subcloning into a shuttle plasmid -> recombination between the shuttle plasmid and the appropriate adenoviral genome backbone -> produce crude adenovirus stock in 293 helper cells -> adenovirus amplification and purification via CsCl / Iodixanol ultra-centrifugation gradient, or by chromatography -> qPCR titration or PFU titration - Technical support for adenovirus using Case Studies  Figure1. Rat astrocytes were infected with Ad5-hHTT at an MOI of 10, 50, and 100. At 24 h after infection, total protein were harvested and processed for western blot analysis. 
Figure2. Comparison of GFP delivery efficiency between plasmid transfection and adenovirus transduction in different cell types. |
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