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Antibody-dependent cell-mediated cytotoxicity (ADCC) is a mechanism by which the Fab segment of an antibody binds to antigenic epitopes on virus-infected or tumor cells, while its Fc segment interacts with Fc receptors (FcR) on effector cells (such as NK cells and macrophages). This interaction mediates the direct killing of target cells by effector cells and represents an important mechanism of action for therapeutic anti-tumor antibody drugs.

  
Traditional methods for measuring ADCC/ADCP primarily rely on the isolation and in vitro differentiation of relevant primary immune cells, followed by the assessment of target cell killing or phagocytosis. These methods are highly dependent on donor-derived primary cells, are costly, time-consuming, and labor-intensive. The isolation and differentiation process of primary cells imposes stringent requirements on experimental operations, increasing the risk of experimental failure. Additionally, due to purity issues with differentiated cells, these methods often suffer from low and unstable detection signals. Furthermore, primary cells are non-renewable, leading to potential inconsistencies in cell donor sources across experiments, resulting in poor reproducibility and possible batch-to-batch variations. Due to these factors, establishing stable and reliable detection methods is challenging in drug development processes requiring high-quality control.

The Jurkat E6.1 Mouse ADCC Bioassay Effector Cell model effectively mimics the in vivo ADCC signaling transduction process. The principle is illustrated in the figure below.

Figure 1. Schematic diagram of the Jurkat E6.1 Mouse ADCC Bioassay Effector Cell model.

Figure 2. Dose response of Anti-hCD20-mIgG2a in Mouse ADCC Bioassay Effector Cell (C3).

Cell Passage Procedures

1.This cell line grows in suspension.
2.Upon receipt, cells should be thawed immediately or stored in liquid nitrogen until use.
3.Before thawing, pre-warm the water bath and culture medium to 37 °C, and prepare a small amount of dry ice.
4.Remove the cryovial from storage and transport it to the cell culture laboratory on dry ice.
5.Rapidly thaw the cells in a 37 °C water bath. Once the cells are completely thawed, spray the cryovial with 70% ethanol for disinfection and transfer it to a biosafety cabinet.
6.Add 10 mL of pre-warmed culture medium into a 15 mL centrifuge tube. Transfer the contents of the cryovial into the tube and centrifuge at 1000 rpm for 5 minutes.
7.Carefully discard the supernatant. Resuspend the cell pellet in 5 mL of pre-warmed culture medium by gentle pipetting. Immediately perform cell counting and adjust the cell density to 3–6 × 10⁵ cells/mL based on the counting results, then transfer the cells into a culture flask.
8.Count the cells every 1–2 days. When the cell density exceeds 1 × 10⁶ cells/mL, passage the cells promptly or add fresh culture medium. Maintain the cell density between 2 × 10⁵ and 1 × 10⁶ cells/mL.

Suspension Cell Cryopreservation Procedure:

1.Collect 8 × 10⁶ cells, centrifuge, and discard the supernatant.
2.Add 1 mL of cell freezing medium (90% FBS + 10% DMSO) and gently pipette to mix thoroughly. Transfer the suspension into a cryovial.
3.Immediately place the cryovial into a controlled-rate freezing container (Nalgene 5100-0001), fill with isopropanol up to the indicated level, and store at −80 °C.
4.After 24 hours, transfer the cryovial to liquid nitrogen for long-term storage.