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Scientific Background
CD19 (also known as Cluster of Differentiation 19, B-lymphocyte surface antigen B4, or CVID3) is a glycoprotein expressed at the surface of B lymphocytes through most phases of B cell maturation. It is strictly required for B cell terminal differentiation. Mutations in the CD19 gene cause severe immune-deficiency syndromes associated with impaired antibody production such as CVID3 (common variable immuno-deficiency 3). The majority of B cell malignancies express normal to high levels of CD19, making it a nearly ideal target for cancer immunotherapy. Blinatumomab, a CD19/CD3 bi-specific T cell engager (BiTE) has been approved for relapsed/refractory B precursor ALL (Acute lymphoblastic leukemia) and CD19 was the target of the first approved CAR-T cell therapy. Studies of CD19 function and expression profiles will continue to broaden our knowledge and support broader applications in cancer therapy.
Product Description
HIV-based · VSV-G-pseudotyped · SIN=YesThe anti-CD19 CAR lentiviruses are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles that are ready to ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses transduce the ScFv portion of anti-CD19 (clone FMC63) linked to 2nd generation CAR (Chimeric Antigen Receptor), containing CD8 hinge and transmembrane domains, 4-1BB and CD3ζ signaling domains. This construct also includes an IRES- enhanced green fluorescent protein (eGFP) sequence downstream of anti-CD19 CAR cassette to facilitate analysis and sorting of transduced cells. Note: This product transduces the same anti-CD19 CAR construct (CD19 ScFv-CD8-4-1BB-CD3ζ) as other available anti-CD19 CAR Lentiviruses (BPS Bioscience #78600, 78601 and 78602), but differ in key aspects. This product contains an eGFP reporter/selection marker so the transduced cells can be sorted based on eGFP expression. Please see table below. atalog # Self-Inactivation (SIN) Selection Marker 78600 no puromycin 78601 yes no 78602 yes puromycin 78775 yes eGFP
Technical Details
| Vector Type | HIV-based, VSV-G-pseudotyped lentiviral vector |
|---|---|
| Payload / Construct | CD19 ScFv-CD8-4-1BB-CD3ζ · eGFP |
| Target Antigen / Gene | CD19 (B-lymphocyte antigen CD19) |
| Signaling / Architecture | CD8 hinge & TM · 4-1BB · CD3ζ |
| Selection Marker | Puromycin |
| Reporter | eGFP |
| Biosafety Level | BSL-2 |
| SIN Vector | Yes |
| Formulation | The lentiviruses were produced from HEK293T cells, concentrated, and resuspended in DMEM. |
| Supplied As | 50 µl of anti-CD19 CAR at a titer ≥ 3x108 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment. |
| Storage | −80°C; avoid repeated freeze-thaw cycles |
| Hazardous Shipping | UN3373 |
Applications
- Positive control for anti-CD19 CAR evaluation in T cells.
- Transduction optimization experiments.
- Generate anti-CD19 CAR-T cells (for research use only, not for therapeutic purposes).
Biosafety & Safety
The lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and integration into the genomic DNA of the target cells. None of the HIV genes (gag, pol, rev) will be expressed in the transduced cells, as they are expressed from packaging plasmids lacking the packing signal. Although the pseudotyped lentiviruses are replication-incompetent, they require the use of a Biosafety Level 2 facility. BPS recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions.
License & Regulatory
This CD19 ScFv-CD8-4-1BB-CD3ζ · eGFP lentivirus is an HIV-based, VSV-G-pseudotyped vector, which confers broad tropism. It can transduce nearly all mammalian cell types, including primary T cells, non-dividing cells, and most established cell lines.
This product requires Biosafety Level 2 (BSL-2) facilities. Although the lentiviral particles are replication-incompetent, all local, institutional, and federal regulations must be observed. Appropriate personal protective equipment must be worn during handling.
Each lot is supplied at ≥3×10⁸ TU/ml. The exact titer value is lot-specific and is provided with each shipment. Functional titer may vary depending on the target cell type.
No. CAR expression is constitutive and does not require Cre recombinase. The CAR construct is driven by the internal promoter and will be expressed immediately after transduction into the target cells.
This lentivirus includes a Puromycin resistance gene, allowing selection of stably transduced cells by treatment with Puromycin at the appropriate concentration.
Store at −80°C immediately upon receipt. Avoid repeated freeze-thaw cycles, as viral titers decrease significantly with each cycle. For long-term storage, aliquot into single-use volumes before freezing. Short-term storage at 4°C (up to 1 week) is acceptable but may result in minor titer loss.
BioHippo offers customization and add-on services for selected products. Options may include:
- Custom formulations: Virus particles can be packaged in alternative formulations upon request (additional fees may apply).
- Custom titers: Higher-titer preparations may be available; contact us for feasibility and pricing.
- Bulk orders: Volume discounts are available for qualifying order quantities.
- Technical support: Our scientific team can assist with protocol optimization and troubleshooting.
For all customization inquiries, please use the contact form or email support@biohippo.com.
