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Viral Vector Production Unit (UPV) Service Request Form for the Production of a Recombinant Viral Vector (VV) Do not fill this area Tech Form Reception Date: Job accepted: Yes No Starting Material : DNA / Virus / n/a Starting Material Reception Date: Accepted by : Date : UPV DNA ID : UPV VIRUS ID: Solic. # Comments : CLIENT INFORMATION 1. Principal researcher Last name: Position: Phone: e-mail: 2. Associate researcher (optional) Last name: Position: Phone: e-mail: 3. Center or Institution Name: Address: First name: Fax : First name: Fax : City: Country: Zip Code: BILLING INFORMATION 1. Billing contact Last name: Phone: e-mail: 2. Billing address Name: Address: City: Country: P.O. # 1: 1 First name: Fax : Zip Code: Purchase Order number or your equivalent ; only if requested by your Institution for the billing. 81914697 1/7 SHIPPING INFORMATION The shipping costs are at the expense of the requester. The viral vectors must be shipped in compliance with the appropriate international transportation regulations. Therefore, a company authorized for the shipment of Biological Substance, Category B, UN3373 in dry ice from Spain to the country of destination must be used, and the requester should have an account with one such company. 1. Shipping contact (if different from the principal or the associate researcher) Last name: First name: Phone: Fax : e-mail: 2. Shipping address (if different from that of the principal researcher's) Address: City: Country: 3. Courier service Company: Zip Code: Account #: RECOMBINANT VIRAL VECTOR REQUEST GUIDELINES Return the completed form by either: o E-mail: [email protected] (please note: in this case, SECTION 5 must be sent by FAX or signed, scanned and sent by e-mail) o FAX: +34 935 814 205. A separate form should be filled out for each viral vector requested. If the request is accepted, the requester will send the starting material (in case he/she is to provide it) to the following address.: UPV- UAB Edifici H, Campus UAB 08193 Bellaterra (Cerdanyola del Vallés) SPAIN Please make sure that the material you send corresponds to that described in the form. Otherwise, we will not be able to initiate the work. The information you provide will be considered confidential and treated as such. It is necessary for the evaluation and eventual completion or your request, as well as for the correct assignment of a biosafety level. Moreover, it allows the characterization of the viral vectors we produce and advise you in case you do not have enough experience in the use of these type of vectors. The UPV will accept or reject your request according to the information provided. We will notify you our decision within 15 days after receiving the form. The UPV will not accept responsibility for any problem or delay derived from the inaccuracy of the information provided. Any additional expense incurred into due to this cause will be charged to the requester. Storage: Viral vectors will be supplied in storage buffer and shipped frozen in dry ice. They should be stored in a freezer upon arrival. Vectors should be stored long term at -80ºC. Avoid repeated freezing/thawing, especially if virus has been diluted, to prevent significant loss of viral titer. 81914697 2/7 SECTION 1 INTENDED USE OF THE VIRAL VECTOR The requested vector… will be used in vitro? Yes No will be used ex vivo? Yes No will be used in vivo? Yes No Target cells/tissue: In case of in vivo use : Species to be used : Way of administration: Please provide a brief and general description of your research project : 81914697 3/7 SECTION 2 DESCRIPTION OF THE RECOMBINANT VIRAL VECTOR Is it a marker vector routinely made in the UPV? If so, specify: Ad5-CMV-Gal Ad5-RSV-Gal Ad5-CMV-GFP Ad5-CMV-RFP Ad5-CMV-Cre AAV2/X-CMV-Gal X = specify CAV2-Cre CAV2-GFP AAV2/X-CMV-GFP X = specify Is it a non-standard vector already made by the UPV? If so, please provide the following information: Name of the VV: Production number (if known): CBATEGFill this shaded area if your request is for a non-marker, new vector Name of the VV: Adenovirus AAV Serotype choose one Starting material : DNA Virus * * If the starting material is an already made adenovirus, please state: The starting material is: Purified viral stock Titer2: PP/ml; IU or pfu/ml; Volume: µl Crude lysate Has it been tested for… …mycoplasm ? …sterility? …RCA ? YES; result of the test: NO UNKNOWN Comments or additional information relevant to the amplification and/or purification of the VV : Requested production scale: Marker Ad, 1x1011 PP aliquot Standard production Number of aliquots: Other Scale choose one Note : Our standard production is one derived from 20 15-cm plates, and it is characterized by the total number of generated viral particles. The average yields are offered as a guide for deciding on the production scale, and are not to be taken as a predictor for the requested VV: - AAVs: 1x1013 viral genomes, with average titer 7E1012 vg/ml - Ads: 4x1012 PP and 3x1011 IU, with average titers of 1,5x1012 PP/ml and 1x1011 IU/ml Additional tests: RCA (only for Ad) Other : Note: we routinely perform Mycoplasm detection and sterility tests in all our productions. Aliquot volumes: 50 l 100 l 200 l Other (>200 µl) : Note: The final stock must be stored at -80ºC and aliquoted in order to minimize the freeze/thaw cycles which lower the titer. We will try to provide the requested aliquot volumes up to a maximum of 50 aliquots. 2 PP, physical particles; IU, Infectious Units. 81914697 4/7 SECTION 3 DESCRIPTION OF THE EXPRESSION CASSETTE Do NOT fill this section if the requested vector is a standard marker or has already been produced by the UPV TRANSGENE is defined here as the DNA sequence already inserted, or to be inserted, in the expression cassette. If the starting material is a recombinant adenovirus, it refers to the foreign DNA sequence expressed from it. Promoter Transgene (name, species of origin) Poly A 1. Biologic activity of the transgene (oncogene, enzyme, structural protein…) : 2. Is it involved in cell growth? Yes No 3. Could its expression result in tumor induction? Yes No 4. Could its expression confer resistance to drugs or antibiotics? Yes No 7. Is there any safety concern regarding the overexpression of the transgene in human Yes beings? No 5. Transgene expression from its natural promoter is: Constitutive in mammalian cells Cell type/tissue specific Other (state) Unknown 6. Regarding cell growth and metabolism, the expression of the transgene…: is toxic is toxic when overexpressed inhibits cell growth has no known effects 8. If the answer to (#7) is YES, explain why: 9. Is the transgene expression product secreted? Yes No 10. Comments/ additional information : 81914697 5/7 SECTION 4 DESCRIPTION OF THE PLASMID TO BE SENT Do NOT fill this section if the requested vector is standard marker or has already been produced by the UPV IF MORE THAN ONE PLASMID IS GOING TO BE SENT, MAKE AS MANY COPIES OF THIS SECTION AS NEEDED - STARTING MATERIAL TO PROVIDE : any plasmid harboring the transgene, purified by any method yielding high quality DNA (A260/A280 1,8); we strongly suggest the use of reliable kits. If a cloning process is to be performed, it is recommended that the transgene expression be checked through transient transfection before the actual VV production is initiated. Plasmid name : Production date : Plasmid size: kilobases Insert size: kilobases Selection marker: Kit or production process : Dissolved in (H20, TE…) : Plasmid concentration: µg/µl (minimum: 0,5 µg/µl) Plasmid amount : µg (minimum: 10 µg) In order for us to design the cloning strategy, you MUST provide at least one of the following : Complete electronic sequence of the whole construct in a text file *, wih a clear indication of the part of the sequence corresponding to the transgene to be subcloned. Yes No Complete electronic sequence of the transgene in a text file *, with a clear indication of the flanking restriction enzymes which would allow its subcloning. Yes No A restriction map of the plasmid with as much information as possible, at the very least including : o Sizes of plasmid and insert o Relevant restriction enzymes locations o (only for Adenoviruses) Pac I sites presence/absence Yes No * Note: the failure to provide this information will make it necessary for the UPV to do additional work, the cost of which will we added to the final price. This would be done with the informed consent of the requester. Comments / additional information which could be useful : * Send the file(s) to the e-mail address : [email protected] 81914697 6/7 SECTION 5 REQUEST VALIDATION DISCLAIMERS - The viral vectors produced by the UPV are for research use only, and not for use in diagnostic of therapeutic applications. - The UPV is a non-commercial entity operating within the Universitat Autònoma de Barcelona. All charges by the UPV are for scientific services or laboratory supplies, and are not a sale of biological materials. - The viral vectors produced are potentially infectious biological agents. By default, they are assigned to Biosafety Level 2 –unless there are reasons for their adscription to a different level. In any case, they should used under appropriate containment and by trained lab personnel. The suggested containment level is determined on the basis of the potential effect of the virus in a healthy human adult; special considerations should be applied to instances in which an individual may be at increased risk of acquiring infection of for whom infection may be unusually hazardous (e.g., preexisting diseases, medications, compromised immunity, pregnancy or breast feeding). - The requester hereby acknowledges his/her full responsibility for following whatever law or regulations may be of application regarding the use of the viral vectors in his/her Institution, as well as for the consequences derived from their misuse. The UPV shall not be held liable for any damage resulting from the handling or use of these products. Date: 81914697 P.I. signature: 7/7