Pre-made and Custom-made Lentivirus vectors (LVVs)
Lentivirus (LV), a single-stranded RNA retroviral vector, is widely used as a gene transfer tool in the development and production of CGT therapies due to its capability to infect both dividing and non-dividing cells.
The traditional approach for lentivirus production relies on an adhesion cell production system in the presence of fetal bovine serum. This system utilizes animal-derived components and presents challenges in scalability. To overcome these factors, a serum-free suspension culture system was created for the lentivirus vector manufacturing process. Through the LV-SMARTTM platform, we provide pre-made and custom-made LVV.
Porton Advanced has developed LV-SMARTTM. a suspension platform that utilizes our proprietary suspension HEK293T cell lines and harvest/purification technologies. This system has been successfully employed to produce LV products with the production of high titer lentivirus.
Services
Porton Advanced employs an approach utilizing the third generation VSV-G lentivirus packaging. This system involves a 4-plasmid setup, employing suspension HEK293T cells that are cultivated in a serum-free and animal component-free medium. For the Standard scale, following incubation, the supernatant is harvested, subjected to centrifugation to eliminate cellular debris, and subsequently filtered. Lentiviral particles are concentrated using PEG.
For the Advanced scale lentivirus production intended for in vivo applications, viral particles undergo additional purification via chromatography and are further concentrated using TFF.
The lentivirus titer is measured using the p24 ELISA method. But we also offer qPCR or FACS titration.
| Scale | Batch Size | Material Description | Expected Titer | Turnaround Time | Price(USD) | Turnaround Time(BD) |
| Standard-Mini | 5 X 10^7 TU | Standard Concentrated Material (In vitro purpose) | >10^8 TU/mL | 500 uL ( 5 vials X 100 UL) | $ 529 | 10-16 BDs |
| Standard-Medium | 1 X 10^8 TU | >10^8 TU/mL | 1 mL( 10 vials X 100 uL) | $ 629 | 10-16 BDs | |
| Standard-Large | 1 X 10^9 TU | >10^9 TU/mL | 1 mL( 10 vials X 100 uL) | $ 1,049 | 15 -20 BDs | |
| Advanced-Medium | 5 X 10^8 TU | Advanced-Purified Concentrated Material (In vivo/In vitro purpose) | >10^9 TU/mL | 500 uL( 10 vials X 50 uL) | $ 1,329 | 15 -20 BDs |
| Advanced-Large | 1 X 10^9 TU | >10^9 TU/mL | 1 mL( 10 vials X 100 uL) | $ 1,599 | 20-22 BDs | |
| Advanced-XLarge | 2 X 10^9 TU | >10^9 TU/mL | 2 mL( 20 vials X 100 uL) | $ 2,099 | 10-16 BDs |
Note: “TU” refers to transduction unit, “ul” refers to microliter, “ml” refers to mililiter,and “BD” refers to business day.Note: Lentivirus transducing units per milliliter (TU/mL) is determined by a p24 ELISA assay. The titer calculation is formed by converting the concentration of p24 to viral titer using the conversion factor from Didier Trono. This conversion factor is approximately 1 x 104 physical particles of lentivirus per pg of p24 antigen. Based on ELISA results, 1 pg of p24 antigen detection contains 100 transducing units (TU).
Workflow
| Day 0 | Seed cells in shake flasks or Bioreactor |
| Day 1 | Transfect cells with 4 plasmids |
| Day 3-4 | Perform Harvest/Clarification, Concentration, and/or Purification/Concentration |
| Day 4-5 | Perform quality control |
Quality control (QC)
Our analytical team will conduct an ELISA-based physical titer assay (P24) and safety tests including sterility and mycoplasma, to safeguard the quality of all LVV products. We guarantee negative detection in mycoplasma testing and offer expedited sterility testing by either the qPCR method or the rapid microbiological method. Infection titer by FACS, endotoxin tests, and additional testing items are available upon request. Our analytical team is well-equipped with a diverse range of instruments, enabling us to offer flexible testing methods and achieve rapid turnaround times.
Deliverables
- Lentivirus Product
- Certificate of Analysis (CoA)
Storage
Our lentivirus can be formulated in an in-house storage buffer (modified PBS buffer) and stored at -80°C. Although this product can be transported on dry ice for domestic and international shipments, it should be stored at -80°C upon receipt for long-term storage. The shelf life for the lentivirus product is approximately one year.
Note: please avoid repeated freeze-thaw cycles.