MMLV Retrovirus Packaging
Recombinant Moloney murine leukemia virus (MMLV) retroviral vectors are an efficient tool for stable gene delivery into dividing cells that came into prominence for for generation of induced pluripotent stem (iPS) cells. VectorBuilder’s proprietary technologies have been optimized and extensively validated for production of high-titer, high-purity MMLV with excellent viability and batch-to-batch consistency.
- Expertise: Our proprietary packaging technology consistently delivers high-titer, high-efficiency virus.
- Streamlined: Design and order your MMLV vector plus packaging in minutes with our highly intuitive Vector Design Studio.
- Customizable: Choose wild-type or self-inactivating VSV-G pseudotyped MMLV, then build your vector or leverage our team’s design expertise.
- Scalable: Reliable production of high-quality virus at various scales, from research- to GMP-grade.
Service Details
Price and turnaround Price Match
Plasmid preparation is included in the virus packaging service.
| Scale | Application | Titer | Volume | Price (USD) | Turnaround |
|---|---|---|---|---|---|
| Pilot | Cell culture | >107 TU/ml | 250 ul (10x25 ul) | $449 |
6-12 days
|
| Medium | 1 ml (10x100 ul) | $649 | |||
| Large | >108 TU/ml | 1 ml (10x100 ul) | $1,099 | ||
| Ultra-purified large | Cell culture & in vivo | >108 TU/ml | 1 ml (10x100 ul) | $1,699 |
TU = Transduction units (aka infectious units)
Deliverables
| For non-ultra-purified scales | For ultra-purified scales |
|---|---|
| Your custom MMLV retrovirus | Your custom MMLV retrovirus |
|
Free: control virus
|
Add-on purchase (optional): ultra-purified control virus
|
| Free: Polybrene (5 mg/ml, 200 ul) | Free: Polybrene (5 mg/ml, 200 ul) |
Control virus
The control MMLV retrovirus vector is designed to mirror the experimental construct, allowing the effects of the custom construct to be distinguished from those associated with transduction and reporter expression. For example, if the custom virus overexpresses a gene, then the control virus provided will be EGFP control MMLV retrovirus (MMLV retrovirus overexpressing EGFP). Detailed information on our control viruses is shown below:
Shipping and storage
Our MMLV retrovirus is stored in HBSS buffer and is shipped on dry ice. Upon receipt, it should be stored at -80°C for long term (stable for at least 6 months), or -20°C for use within one week. The shelf life of MMLV retrovirus is approximately one year. Please avoid repeated freeze-thaw cycles, as this can result in a large titer drop.
Technical Information
MMLV retrovirus production and quality control (QC)
For our MMLV retrovirus manufacturing, the transfer plasmid carrying the gene of interest (GOI) is co-transfected with our proprietary envelope plasmid encoding VSV-G and packaging plasmid encoding Gag/Pol into packaging cells. After a short incubation period, the supernatant is collected and centrifuged to remove cell debris and then filtered. Viral particles are subsequently concentrated with PEG. For ultra-purified MMLV retrovirus (in vivo grade), viral particles are further purified and concentrated by sucrose gradient ultracentrifugation. We use a qPCR-based approach to measure MMLV functional titer.
For each recombinant MMLV retrovirus 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 perform a transduction test to detect corresponding fluorescence. Additionally, for ultra-purified MMLV, we routinely check endotoxin levels. Including the endotoxin results in your COA incurs an extra fee. Additional QC services can also be provided upon request.
How to Order
Order both vector cloning and virus packaging
Order virus packaging of your own vector
If customer-supplied MMLV retroviral plasmids are needed, 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.
Resources
FAQ
What features does MMLV retrovirus have comparing to other viral vectors?
| Lentivirus | MMLV | Adenovirus | AAV | |
|---|---|---|---|---|
| Tropism | Broad | Broad | Ineffective for some cells | Depending on viral serotype |
| Can infect non-dividing cells? | Yes | No | Yes | Yes |
| Stable integration or transient | Stable integration | Stable integration | Transient, episomal | Transient, episomal |
| Maximum titer | High | Moderate | High | Very high |
| Promoter customization | Yes | No | Yes | Yes |
| Primary use | Cell culture and in vivo | Cell culture and in vivo | In vivo | In vivo |
| Immune response in vivo | Low | Low | High | Very low |
What is VectorBuilder's guarantee for virus titer and turnaround?
Our titer guarantee only applies to vectors for which the region being packaged into virus (from 5’ MoMuLV LTR to 3’ MoMuLV LTR) is below MMLV cargo limit (8.3 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 MMLV functional elements (e.g. LTR) that may introduce uncertainties to packaging efficiency.
Our estimated turnaround is the time from production initiation to completion. It does not include waiting time for any customer-supplied materials (e.g. template DNA or viral vectors), QC of such materials, and transit time for shipping final deliverables to the customer.
