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ADCP (Antibody-Dependent Cellular Phagocytosis) is antibody-dependent cellular phagocytosis, an immune elimination mechanism that targets tumor cells with monoclonal antibodies (mAb) to promote the clearance of tumor cells from the body by phagocytic immune cells. ADCP is mediated by monocytes, macrophages, neutrophils, and dendritic cells through the expression of FcγRIIa (CD32a), FcγRI (CD64), and FcγRIIIa (CD16a) to phagocytose diseased cells. Studies have shown that FcγRIIa is the FcγR receptor involved in this process.

  
Traditional methods for determining ADCC/ADCP primarily rely on the isolation and in vitro differentiation of relevant primary immune cells, followed by measurement of target cell killing or phagocytic effects. These methods are highly dependent on donor primary cells, are costly, time-consuming, and labor-intensive. The isolation and differentiation process of primary cells imposes very high requirements on experimental operations, increasing the risk of experimental failure. Additionally, due to purity issues with isolated and differentiated cells, there are problems of low detection signals and instability. Furthermore, primary cells are non-renewable, and the cell donor sources for each experiment may be inconsistent, leading to poor reproducibility of experimental results and potential batch differences. Due to these factors, it is difficult to establish stable and reliable detection methods in drug development processes requiring high-quality control.

The Jurkat E6.1 Human ADCP Bioassay Effector Cell FcγRIIa (H variant) -NFAT Cell model effectively mimics the in vivo MET signaling transduction process. The principle is illustrated in the figure below.

Figure 1. Schematic diagram of the Jurkat E6.1 Human ADCP Bioassay Effector Cell FcγRIIa (H variant) -NFAT Cell model.

Figure 2. Dose Response of Rituximab in ADCP Bioassay Effector Cell FcγRIIa (H variant)-NFAT Jurkat(C2) with Raji.

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.