Biosafety training and registration are required when you wish to work in the BVC facility. Please contact our customer support firstname.lastname@example.org to schedule the training. The biosafety training takes roughly 1-1.5h during which we go through the most significant biosafety issues and take a quick tour in our facility.
Please note that the biosafety training is only about how to use the facility safely. We do not provide protocols or guidance for the actual virus work. If you’re new to virus work, we recommend organising your own supervisor or someone else to train you for the protocols and habits. The supervisor/teacher/trainer also need to be a registered BVC user.
After a customer has been trained and registered as a BVC facility user (STEMM users included) they are able to book the laminar hood for their experiments. All the facility instruments are free to use without extra fees. BVC users are also free to use all the cell culture plastics and reagents found in the facility. STEMM users need to provide their own accessories, plastics and reagents. Ethanol, MQ water, Virkon, pipettes and pipette boys are free for everyone.
Recombinant lentiviruses are designed to perform only one cycle of infection. After the transduction, a segment of the transfer vector that encodes shRNA/miRNA or cDNA integrates into the host cell's genome and is expressed under the control of the viral promoter. The recombinant viruses do not carry genetic information for production of virus particles and, therefore, cannot replicate or produce new virus particles in the target cells. However, there is a small possibility that unusual recombination events lead to formation of replication competent viruses during the virus production. The presence of replicating competent virus (RCV) is excluded in lentivirus preparations with the RCV test.
RCV test is performed to transduced cells to determine that the lentiviral vector is not replicating and to ensure the cells are virus-free before transferring cells from a BSL-2 laboratory a BSL-1 facility. We also recommend to always test tranduced cells for replicating viruses. NOTE: We do not test primary cells or tissues for wild type HIV-1!
A 24 kDa capsid protein (p24) is a major structural core component in the recombinant lentiviruses. We use a p24 specific enzyme-linked immunosorbent assay (ELISA) to determine the p24 lentivirus capsid protein concentration (pg/ml) from the cell culture supernatants containing lentivirus preparations.
These tests are performed every other Wednesday and the test schedule can be found from the BVC Online Hood reservation calendar. For RCV test you need to provide us 1 ml of unfiltered cell culture media. The media sample should be collected from a transduced cells after at least three passages, and the media should be kept on the cells at least 24 hours. If you are collecting the RCV test from suspension cell culture, remember to pellet and wash your cells with PBS during each passaging. For p24 titer analysis 10 μl of viral supernatant is needed. Inform the customer service whether your lentivirus media is concentrated or not, as this effects the sample preparation and final results. Label samples with your initials and running numbers (EN1, EN2…ENn). If you are a registered BVC facility user, you can deliver your samples in the -20°C sample box at the BVC facility. You can also contact our customer service to arrange the sample delivery. Remember to fill the order form and send an email email@example.com in order to have your samples analysed!
Please sote that the p24 capsid protein based tests are not suitable for retroviruses.
We produce lentivirus particles from customer constructs. Customers are welcome to deliver their lentiviral plasmids to us for custom particle production.
We also produce lentivirus from TRC1 shRNA and the Sanger CRISPR knockout gRNA libraries overseen by the Genome Biology Unit. The clones are available as glycerol stocks, plates, DNA minipreps or midipreps. The library products are produced and distributed by GBU. To inquire more about these libraries, clones and GBU services please contact firstname.lastname@example.org. For more information about the TRC and CRISPR libraries, please visit the GBU web page.
NOTE: For lentivirus production HelVi-BVC uses second generation packaging system and therefore we also use wild type LTR viral promoter. Second generation packaging system and wild type viral promoter do not automatically create replicating lentivirus but there is a higher risk for this to happen than with third generation packaging system. Therefore, due to safety reasons, we highly recommend always testing your transduced cells lines for replicating lentivirus.
Each lentivirus production process includes p24 titer test to determine the quality and concentration of the lentivirus particles. In addition, we use an internal control sample to evaluate the virus quality.
Here are presented the expected p24 titer concentration (pg/ml) ranges between different lentivirus products:
Please note: The p24 and internal control analyses are for lentiviruses only. For retroviruses, quality control tests depend on the vector backbone and type (ecotropic or amphotropic pseudotypes). Please contact email@example.com for more information. Unfortunately HelVi-BVC doesn’t currently offer titer tests for retrovirus.
When you’re providing your own plasmids for virus production, here’s how much DNA you need to provide:
Please contact us for a quote request! We are happy to help you find the viral product most suitable to your needs!
HelVi-BVC also produces retrovirus particles from customer DNA.
Our retrovirus products:
Required customer plasmid DNA:
At the moment HelVi-BVC doesn’t provide titer or replicating competent virus testing for retrovirus.
If you need to concentrate and purify your lentivirus supernatant, we can do this for you in HelVi-BVC by ultracentrifugation.
The lentivirus supernatant is pelleted and purified in a sucrose medium by ultracentrifugation and then resuspended in the volume of interest.
The minimum volume of lentivirus media required for concentration is 37 ml.
For further information please contact our customer support: firstname.lastname@example.org.
We also provide cell lines for virus production: HEK293 FT, Phoenix Amphotropic, Phoenix Ecotropic and HeLa cells.
Please note! It is the user’s responsibility to test the cells for absence of mycoplasma. In addition, the passage numbers of the cells vary between 7-10+, and we can’t promise a cell line with low passage numbers (1-5).
The HelVi-BVC research infrastructure is supported by University of Helsinki (HiLIFE and Research Programs Unit) and Biocenter Finland. The Biomedicum Virus Core price list is presented below. Additional prices, where given, apply to subsequent products or services of the same type within the same order. The prices are applied for three different customer groups:
For further information please contact email@example.com.
|Biosafety training and registration
|Concentrated lentivirus,180 ul
|Concentrated lentivirus, 380 ul
Sucrose purification of concentrated lentivirus
Please note: Plasmid preparation or validation are not included in the lentivirus production services! Customers are expected to provide their own plasmids for lentivirus production. Plasmids from the former FuGU Libraries TRC and CRISPR libraries are prepared and distributed by the GBU core. The GBU's plasmid preparation fees are charged separately and those are not included in the virus production prices presented above. For further information please contact GBU customer service or visit their web page.