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Application: Expression of human MAGE-A4 (p230-239) B2M HLA-A*02:01 in cells of interest.Generate MAGE-A4 (p230-239) B2M HLA-A*02:01 expressing cell pools or stable cell lines by puromycin selection.
Background: Human Leukocyte Antigen-A (HLA-A) is an MHC-I (major histocompatibility complex) heavy chain receptor, composed of HLA-A and β2-microglobulin (B2M). There are over 200 genes encoding HLA variants and this variability plays a critical role in adaptive immunity. HLA-A*02 is one of the most common class I types. HLA class I are involved in presenting peptides that are typically between 8 to 11 amino acids. HLA-A*02:01 can bind 15-mer peptides, which can then be recognized by T cells. Studies in SCLC (small cell lung cancer) patients has shown that an ATAD2 (ATPase family AAA domain-containing protein 2) immunopeptide can be used in HLA-A*02-01-restricted patients with high reactivity. 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. In 2024, the first MAGE-A4 TCR engineered cell therapy for advanced synovial sarcoma was approved by the Food and Drug Administration (FDA).
Description: MAGE-A4 (p230-239) B2M HLA-A*02:01 Lentivirus are replication incompetent, HIV-based, VSV-G pseudotyped lentiviral particles ready to transduce most mammalian cells, including primary and non-dividing cells. These viruses result in the expression of MAGE-A4 (melanoma associated antigen A4) peptide 230-239, B2M (beta-2 microglobulin), and HLA-A (human leukocyte antigen) *02:01. The lentiviruses also contain a puromycin selection marker (Figure 1). A B Figure 1. (A) Schematic of the lenti-vector used to generate MAGE-A4 (p230-239) B2M HLA-A*02:01 Lentivirus. (B) Construct diagram showing components of the MAGE-A4 (p230-239) B2M HLA-A*02:01.
Formulation: The lentivirus particles were produced in HEK293T cells. They are supplied in medium containing 90% DMEM + 10% FBS. Virus particles can be packaged in custom formulations and produced at higher titers by special request, for an additional fee.
Storage Stability: Lentiviruses are shipped with dry ice. For long-term storage, it is recommended to store the lentiviruses 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.
Supplied As: Two vials (500 µl x 2) of lentivirus at a titer ≥107 TU/ml. The titer will vary with each lot; the exact value is provided with each shipment.
Target: B2M, HLA
Uniprot: P61769, P04439
Warnings: BiosafetyThe lentiviruses are produced with a SIN (self-inactivation) lentivector which ensures self-inactivation of the lentiviral construct after transduction and after integration into the genomic DNA of the target cells. 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 and are not present in the lentivirus particle. 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.Avoid freeze/thaw cycles.
Biosafety Level: BSL-2
SIN Vector?: Yes
References: Kropp KN., et al., 2020 PLOS One 15(9): e0238875.Caballero, O L.., et al., 2009 Cancer Sci. 100, 2014–2021.Sanderson, J. P. et al., 2020 Oncoimmunology 9, 1682381.Hassan C., et al., 2014 J Biol Chem 290(5):2593-2603. Yuan L., et al., 2025 eBioMedicine 112: 105515.
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