The Laboratory of Prof. dr. Kyriakos E. Kypreos at the University of Patras Department of Medicine in Greece provides investigators from all over the world with comprehensive service in the creation of high quality attenuated recombinant adenoviruses. A team of experts with long presence in the field of recombinant adenovirus construction and purification, under the direct supervision of Dr. Kypreos, will make sure that your adenoviruses will be of the highest possible titer and purity for your in vitro and in vivo applications. For your convenience, the recombiant attenuated adenovirus will express:

• Your Favorite Gene (YFG) under the control of a CMV promoter
• The Green Fluorescent Protein (GFP) under the control of an independent CMV promoter. 

Fees

Customers provide the gene of intertest cloned in the appropriate carrier vector and in return our core facility will supply the investigator with a purified recombinant adenovirus. 

For amplification, purification, and titration of the adenovirus the expected project duration is 4 weeks. The fee for this service is €3,000 per viral preparation.
 
For generation, amplification, purification and titration of the adenovirus, the expected project time is 8-10 weeks. The fee for this service is €7,000 per viral preparation.

Investigators should also provide an account number of the courier of their choice for the shipment of the adenovirus.

How to order

To initiate the requested service, please express your interest in generating an adenovirus by submitting and email request to Dr. Kyriakos Kypreos (kkypreos@med.upatras.gr). Following confirmation of your order you will be requested to pay the fee in full using one of the following bank accounts:

National Bank of Greece

Account No: 22954000232
IBAN: GR6001102290000022954000232
Swift Code: ETHNGRAA

Alpha Bank

Account No: 543002001000029
IBAN: GR1101405340534002001000029
Swift Code: CRBAGRAAAXXX

To easily track your payment, when making the wire transfer please use the following elements in the justification field:

Prof. dr. K Kypreos Laboratory, Adenovirus Facility, Account C.699

For the generation of a new adenovirus: Following receipt of the order form and full payment, our facility will then provide an adenovirus shuttle plasmid map and a “project number” to the investigator. The investigator is responsible for sucloning the gene of interest and screening the construct in order to confirm that the correct construct has been made.)  The investigator will then return the adenovirus shuttle plasmid to the facility in sufficient quantities (200 μg). A copy of the order form along with the “project number” must accompany the shuttle plasmid.  The newly created adenoviral vector will be grown at a large scale and purified using double CsCl ultracentrifugation and dialysis.

For the amplification of an existing adenovirus: Following receipt of the order form and full payment, our facility will then provide a “project number” to the investigator. Investigators who have already an adenovirus and need only the amplification, purification and titration services should should ship an aliquot of the adenovirus on dry ice to the core facility. A copy of the order form along with the “project number” must accompany the adenovirus aliquot.    

Considerations for cloning

1) The insert of interest that is cloned into the shuttle vector should not contain the restriction sites Pme I and Pac I. Please avoid cloning your insert in these sites.

2) Please avoid cloning elements that are present more than once in the vector (e.g., CMV promoters) in head-to-head orientations.

3) Please note that all the shuttle vectors confer resistance to Kanamycin (not ampicilin).

Do you need on-site help with in vivo administration of adenovirus to mice or rats via the tail-vein? 

Do you need to use your adenovirus for in vivo hepatic expression of YFG and don't know how? Our facility experts can visit your laboratory and perform in vivo administration of adenoviruses via tail vein to the mouse and rat model of your choise. Custom rates apply. Please inquire by email.  

References

1. He et .al.,  (1998) A simplified system for generating recombinant adenoviruses, Proc. Natl. Acad. Sci U.S.A. 95: 2509-2514.

2. Kypreos KE (2008) ABCA1 promotes the de novo biogenesis of apolipoprotein CIII-containing HDLparticles in vivo and modulates the severity of apolipoprotein CIII-induced hypertriglyceridemia, Biochemistry, 47: 10491-502.

3. Kypreos KE et. a., (2007) Pathway of biogenesis of apolipoprotein E-containing HDL in vivo with the participation of ABCA1 and LCAT, Biochem J, 403: 359-67. 

4. Kypreos KE, et. al., (2005) Generation of a recombinant apolipoprotein E variant with improved biological functions: hydrophobic residues (L261, W264, F265, L268, V269) of apoE can account for the apoE induced hypertriglyceridemia J.Biol.Chem., 280:6276 - 6284.