Lentivirus

Recombinant lentivirus is perhaps the most commonly used vectors for efficient gene delivery into mammalian cell culture. Unlike conventional plasmid transfection which only brings foreign DNA into cells transiently, lentiviral vectors can introduce genes into cells permanently through genomic integration. Lentivirus is also used for in vivo gene delivery applications.
VectorBuilder offers superior quality lentiviral packaging services to support your lentivirus-based gene therapy experiments. We have developed a series of proprietary technologies and reagents that have greatly improved recombinant lentivirus production protocols in terms of titer, purity, viability and consistency, especially for the third-generation lentiviral vector systems used in our vector construction services. As a result, we have a growing base of highly satisfied customers who come back to us time after time for their lentiviral vector cloning and lentivirus packaging needs.
Click here to read detailed introduction of our lentiviral vector systems
Click here to view citations of our lentivirus
Click here to view user testimonials of our lentivirus
How do I obtain lentivirus packaging services?

Click here to view an example of a finished lentiviral vector design
Click here to view an example of a lentivirus service proposal
Click here to view instructions on designing a vector on VectorBuilder
Click here to view instructions on placing an order on VectorBuilder

Types of lentivirus offered

We offer packaging services for VSV-G pseudotyped second- and third-generation lentivirus. We can also package integrase-defective lentivirus (IDLV).

Price and turnaround

Due to our technology advantage, our lentivirus packaging services have the highest quality in the industry and are also faster and more affordable than those from other suppliers. Prices and turnaround times for different scales of lentivirus packaging are shown below:

Scale Application Titer & Volume* Price (USD) Turnaround**
Pilot Cell culture >108 TU/ml, 10x 25 ul, HBSS buffer $399 8-16 days
Medium Cell culture >108 TU/ml, 10x 100 ul, HBSS buffer $599 8-16 days
Large Cell culture >10TU/ml, 10x 100 ul, HBSS buffer $999 8-16 days
Ultra-purified medium Cell culture & in vivo >109 TU/ml, 10x 50 ul, HBSS buffer $1199 8-16 days
Ultra-purified large Cell culture & in vivo >109 TU/ml, 10x 100 ul, HBSS buffer $1499 8-16 days

TU = Transduction unit (aka infectious unit)

* Our titer guarantee applies to vectors for which the region being packaged into virus (from Δ5’ LTR to ΔU3/3’ LTR) is below the lentivirus cargo limit (9.2 kb). For sizes above this, it may still be possible to package the vector into virus, but the titer may be reduced. Additionally, we are not able to guarantee titer for the following vectors:

  • vectors containing sequences that could adversely affect the packaging process such as toxic genes (e.g. proapoptotic genes), genes that compromise the integrity of packaging cells or virus (e.g. membrane proteins that cause cell aggregation), and sequences prone to rearrangements or secondary structures (e.g. repetitive or highly GC-rich sequences);
  • customer-supplied plasmids containing undisclosed sequences or atypical lentiviral functional elements (e.g. LTR) that may introduce uncertainties to packaging efficiency.

** Turnaround time is defined as the time from when an order is accepted to when it is shipped. It does not include waiting time for any materials that the customer needs to supply (e.g. viral vectors) and QC of the customer-supplied materials.

Deliverables

If you choose pilot, medium or large-scale lentivirus packaging, each order includes the following deliverables:

  • Aliquots of custom lentivirus at selected scale
  • Free: aliquots of control virus*
  • Free: Polybrene (5 mg/ml, 200 ul)

If you choose ultra-purified medium or ultra-purified large-scale lentivirus packaging (in vivo grade), each order includes the following deliverables:

  • Aliquots of custom lentivirus at selected scale
  • "Add-on" purchase (optional): aliquots of ultra-purified control virus*
  • Free: Polybrene (5 mg/ml, 200 ul)
* The control lentivirus is designed to match the biological application of the custom virus and to be used for testing lentiviral transduction. For example, if the custom virus overexpresses a gene, then the control virus provided will be EGFP lentivirus (lentivirus overexpressing EGFP), and if the custom virus expresses an shRNA against a gene, then the control virus provided will express a scramble shRNA. Detailed information for the various control viruses is shown below:
Vector System Control Vector Name Control Vector ID Custom Virus Packaging Scale Corresponding Control Virus Amount Price (USD)
Lentivirus gene expression system

pLV[Exp]-EGFP:T2A:
Puro-EF1A>mCherry

VB160109-10005 Pilot >108 TU/ml, 100 ul in HBSS Free
Medium >108 TU/ml, 2x 100 ul in HBSS Free
Large >108 TU/ml, 2x 100 ul in HBSS Free
Ultra-purified medium >109 TU/ml, 10x 50 ul in HBSS $400
Ultra-purified large >109 TU/ml, 10x 100 ul in HBSS $500
Lentivirus U6-based shRNA knockdown system

pLV[shRNA]-EGFP:T2A:
Puro-U6>Scramble_shRNA

VB151023-10034 Pilot >108 TU/ml, 100 ul in HBSS Free
Medium >108 TU/ml, 2x 100 ul in HBSS Free
Large >108 TU/ml, 2x 100 ul in HBSS Free
Ultra-purified medium >109 TU/ml, 10x 50 ul in HBSS $400
Ultra-purified large >109 TU/ml, 10x 100 ul in HBSS $500
Lentivirus miR30-based shRNA knockdown system

pLV[miR30]-EGFP/Puro-EF1A>mCherry:
Scramble_miR30-shRNA

VB180124-1174nde Pilot >108 TU/ml, 100 ul in HBSS Free
Medium >108 TU/ml, 2x 100 ul in HBSS Free
Large >108 TU/ml, 2x 100 ul in HBSS Free
Ultra-purified medium >109 TU/ml, 10x 50 ul in HBSS $400
Ultra-purified large >109 TU/ml, 10x 100 ul in HBSS $500
Lentivirus CRISPR system

pLV[Exp]-EGFP:
T2A:Puro-EF1A>mCherry

VB160109-10005 Pilot >108 TU/ml, 100 ul in HBSS Free
Medium >108 TU/ml, 2x 100 ul in HBSS Free
Large >108 TU/ml, 2x 100 ul in HBSS Free
Ultra-purified medium >109 TU/ml, 10x 50 ul in HBSS $400
Ultra-purified large >109 TU/ml, 10x 100 ul in HBSS $500
Production, purification and titration

For our third-generation lentivirus manufacturing, transfer plasmid carrying the gene of interest (GOI) is co-transfected with our proprietary envelop plasmid encoding VSV-G and packaging plasmids encoding Gag/Pol and Rev into packaging cells. After a short incubation period, the supernatant is collected and centrifuged to remove cell debris and then filtered. Lentiviral particles are subsequently concentrated with PEG. For ultra-purified lentivirus (in vivo grade), viral particles are further purified and concentrated by sucrose cushion centrifugation. We use the p24 ELISA method to measure lentivirus titer.

Read more about our titration method

Quality control

For each recombinant lentivirus produced by VectorBuilder, quality control includes titer measurement, sterility testing for bacteria and fungi, and mycoplasma detection. If the transfer vector encodes a fluorescent protein, we would transduce cells with virus and detect corresponding fluorescence. Additionally, for ultra-purified lentivirus, we routinely check virus quality using the endotoxin assay.

Customer-supplied vectors

If customer-supplied lentiviral plasmids are used in packaging, please send them to us following the Materials Submission Guidelines. Please strictly follow our guidelines to set up shipment to avoid any delay or damage of materials. All customer-supplied materials undergo mandatory QC by VectorBuilder which may incur a $100 surcharge for each item. Please note that production may not be initiated until customer-supplied materials pass QC.

Questions

Please refer to our Vector FAQ for commonly asked technical questions. To get additional information about what we can offer, please contact VectorBuilder's service team following the contact information here.

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