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PRAME is a protein abnormally expressed in cancer cells, also known as a cancer-testis antigen. It has applications in the diagnosis of melanoma, where it serves as a valuable biomarker. PRAME is abnormally overexpressed in various malignancies, including melanoma and leukemia; it promotes tumor cell proliferation and survival by inhibiting apoptotic pathways (e.g., by interfering with p53 function) while simultaneously helping tumors evade immune surveillance. HLA-A*0201 is a common allele of the Class I Human Leukocyte Antigen (HLA-I) family, responsible for presenting endogenous antigens—such as tumor-associated antigens—to CD8+ T cells, thereby triggering specific immune-mediated killing. When PRAME antigenic peptides are presented by HLA-A*0201, T cells are activated to recognize and attack tumor cells. Studies have demonstrated that cancer patients carrying the HLA-A*0201 allele exhibit higher response rates to PRAME-targeted therapies (such as CAR-T cell therapy and peptide vaccines); consequently, this complex has emerged as a critical target in tumor immunotherapy.

The HLA-A*0201-PRAME complex is a molecular assembly comprising a Class I Human Leukocyte Antigen (HLA) molecule and a PRAME antigenic peptide; it belongs to the Class I family of the Major Histocompatibility Complex (MHC). This complex is frequently utilized in T-cell receptor (TCR) therapies to activate T cells, enabling them to recognize and eliminate tumor cells; the underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the CHO-K1 Human PRAME(SLLQHLIGL) HLA0201 Cell Model

Figure 2. Dose Response of Brenetafusp (IMC-F106C)in NFAT-Luc Jurkat( C7C17) with or without PRAME(SLLQHLIGL) HLA0201 CHO.

Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed  CHO-K1 Human PRAME(SLLQHLIGL) HLA0201 Cell complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.

Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.

2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.

Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.