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Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) refers to a process in which the Fab fragment of an antibody binds to antigenic epitopes on virus-infected cells or tumor cells, while its Fc fragment binds to Fc receptors (FcRs) on the surface of effector cells (such as NK cells, macrophages, etc.). This interaction mediates the direct killing of target cells by the effector cells; it constitutes a crucial mechanism of action for therapeutic anti-tumor antibodies.

Traditional methods for assessing ADCC and ADCP primarily rely on the isolation and in vitro differentiation of relevant primary immune cells, followed by the measurement of target cell killing or phagocytosis. These methods are heavily dependent on primary cells derived from donors; they are expensive, time-consuming, and labor-intensive. Furthermore, the isolation and differentiation of primary cells impose extremely stringent requirements on experimental technique, leading to a significant risk of experimental failure. Moreover, issues regarding the purity of the isolated and differentiated cells often result in low detection signals and a lack of stability and uniformity in the assay results. Additionally, primary cells are non-renewable resources; since the cell donor source may vary from one experiment to the next, the reproducibility of experimental results is often poor, and batch-to-batch variability is a common issue. Due to the influence of these factors, it remains challenging to establish stable and reliable assay methods within the context of drug development, where rigorous quality control is essential.

The ADCC Bioassay Effector Cell Fc Variant (Low Affinity)—Fcγ-NFAT Jurkat Reporter Gene Drug Target Model—effectively simulates the signal transduction process of ADCC *in vivo*. The underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the ADCC Bioassay Effector Cell F variant (Low Affinity)—Fcγ-NFAT Jurkat Cell Model

Figure 2. Recombinant ADCC Bioassay Effector Cell F variant (Low Affinity)- Fcγ-NFAT/Jurkat expressing CD16.

Figure 3. Dose response of CD20 Ab in ADCC Bioassay Effector Cell F variant (Low Affinity)-Fcγ-NFAT Jurkat (C11) 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.