MAGE-A4 Specific TCR Lentivirus

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78935

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Description

MAGE-A4-Specific TCR Lentivirus are replication incompetent, HIV-based, VSV-G-pseudotyped lentiviral particles ready to transduce nearly all types of mammalian cells, including primary and non-dividing cells. These viruses transduce cells with a TCR (T cell receptor) that specifically recognizes the human antigen MAGE-A4 (Melanoma-associated antigen 4) peptide 230-239 (GVYDGREHTV), and in which the TCR α chain and β chain are linked by P2A. The lentiviruses also transduce a puromycin selection marker (Figure 1).

Figure 1. (A) Schematic of the lenti-vector used to generate the MAGE-A4-Specific TCR Lentivirus and (B) diagram of the construct, showing the components of the MAGE-4-specific TCR.
TRAV and TRAC correspond to the TCR alpha chain variable and constant regions, respectively, whereas TRBV and TRBC correspond to the TCR beta chain variable and constant regions

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Synonyms

Cancer/testis antigen 1.4 (CT1.4), MAGE-4 antigen, MAGE-41 antigen, MAGE-X2 antigen, Melanoma-associated antigen 4

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Product Data Gallery

Expression of MAGE-A4-specific TCR in Jurkat cells transduced with the MAGE-A4-Specific TCR Lentivirus.

Jurkat T cell activation after transduction with MAGE-A4-Specific TCR Lentivirus, using T2 cells as Antigen-presenting cells (APCs).

Product Info

Storage and Usage

Citations

Supplied As

≥2 x 10⁷ TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.

Materials Required But Not Supplied

Name	Ordering Information
Thaw Medium 2	BPS Bioscience #60184
Growth Medium 2C	BPS Bioscience #79592
Assay Medium 2D	BPS Bioscience #78755
CD8⁺ TCR Knockout NFAT-Luciferase Reporter Jurkat Cell Line	BPS Bioscience #78757
T2 Cell Line	ATCC #CRL-1992
MAGE-A1 Peptide (278-286)	BPS Bioscience #78965
MAGE-A4 Peptide (230-239)	BPS Bioscience #78966
PE anti-human α/β T Cell Receptor Antibody	BioLegend #306707
ONE-Step™ Luciferase Assay System	BPS Bioscience #60690
Lenti-Fuse™ Polybrene Viral Transduction Enhancer	BPS Bioscience #78939

Formulation

The lentiviruses were produced in HEK293T cells in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations by special request, for an additional fee.

UniProt #

P43358

Background

MAGE (melanoma associated antigen) proteins are CT (cancer testis) antigens, and there are about 60 proteins in the MAGE family that can be subdivided into type I (present only on the X-chromosome, MAGE-A, B and C) and type II (MAGE D-L and necdin). Under normal conditions they are mostly found in the testis and placenta. They are found at high levels in several cancer types, such as melanoma, brain, and breast cancer, and are involved in the development of resistance to chemotherapy, cell motility and cell survival. Expression of MAGE proteins tend to correlate with a poor prognosis. They are intracellular proteins, with MAGE-A4 being found in the cytosol and nucleus, making them poor targets for strategies such as CAR-T cell therapy. MAGE proteins are degraded in the proteosome, and the peptides created can then be found on the cell membrane in combination with MHC (major histocompatibility complex) I. The presentation on the cell surface in this form makes them an attractive target for TCR (T cell receptor)-T cell therapy. Several clinical trials are ongoing, and either alone or in combination with other forms of cancer therapy the use of TCR-T cells targeting MAGE-A antigens may become a promising therapeutic avenue.

References

David O., et al., 2023 Nat Medicine 29:104-114.
De Rooji M., et al., 2023 Molecular Therapy Oncology 28:1-14.

Storage/Stability

Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the virus at -80°C for up to 12 months from date of receipt. Avoid repeated freeze-thaw cycles. Titers can drop significantly with each freeze-thaw cycle.

Growth Media

For the best results, the use of BPS Bioscience validated and optimized media is highly recommended. Other preparations or formulations of media may result in suboptimal performance.

Media Required for Maintaining CD8⁺ TCR Knockout NFAT-Luciferase Reporter Cell Line
Growth Medium 2C (BPS Bioscience #79592): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin plus 1 mg/ml Geneticin, 100 µg/ml Hygromycin B.

Media Required for Maintaining T2 Cell Line
Thaw Medium 2 (BPS Bioscience #60184): RPMI 1640 medium supplemented with 10% FBS, 1% Penicillin/Streptomycin.

Media Required for Co-culture Assay
Assay Medium 2D (BPS Bioscience #78755): RPMI 1640 medium supplemented with 1% FBS.

Applications

Use as a positive control for MAGE-A4 TCR evaluation and optimize experimental conditions.
Generate MAGE-A4 TCR expressing cell pools or stable cell lines, following puromycin selection.

Shipping Temperature

-80°C

Notes

To generate a MAGE-A4 TCR expressing stable cell line, remove the growth medium 48 hours after transduction and replace it with fresh growth medium containing the appropriate amount of puromycin (as pre-determined from a killing curve, https://bpsbioscience.com/cell-line-faq), for antibiotic selection of transduced cells, followed by clonal selection. 

Troubleshooting Guide
Visit bpsbioscience.com/lentivirus-faq for detailed troubleshooting instructions. For further questions, please email [email protected].

Biosafety
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 Bioscience recommends following all local federal, state, and institutional regulations and using all appropriate safety precautions

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cancer Therapy Target

Regulatory Status

Research Use Only

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MAGE-A4 Specific TCR Lentivirus

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