Antibody-dependent cell-mediated phagocytosis (ADCP) is an important mechanism of action (MOA) of therapeutic antibodies. In vivo, ADCP can be mediated by monocytes, macrophages, neutrophils and dendritic cells through FcyRIIa, FcyRI and FcyRIIIa. In terms of mechanism, antibodies bind their homologous antigens to target cells through their antigen recognition domains, and then recruit phagocytes to targets with their Fc regions. Once bound to the Fc receptors of phagocytes, the target cells are taken up and degraded. This process also causes the effector cells to produce soluble factors, which help initiate and drive the immune response. ADCP function is very important in the development of therapeutic antibodies, especially for biosimilars.
Creative Biolabs has extensive experience with ADCP assays and can help you understand the interaction between your compound or biologic and the phagocytic cells of the immune system, taking you to clinic faster. We conduct ADCP bioassays on behalf of clients, either as a standalone project or as part of a larger development project. We are committed to developing the most appropriate experiments to help you understand the molecule.
ADCP can be mediated by FcyRIIa, FcyRI and FcyRIIIa by monocytes, macrophages, neutrophils and dendritic cells. Although all three receptors can participate in ADCP, it is believed that FcγRIIa is the main Fcγ receptor involved in this process.
In our ADCP analysis, the FcγRIIa-H ADCP reporter bioassay is a biologically relevant MOA-based assay that can be used to measure the potency and stability of antibodies and other biological agents that specifically bind and activate FcγRIIa. The assay consists of Jurkat cells stably expressing human FcγRIIa-H (high affinity H131 variant) and NFAT-induced luciferase. The activation of FcγR and NFAT-mediated luciferase activity in effector cells was measured by flow cytometry. Phagocytosis can also be confirmed using microscopy or imaging cytometry.
Fig.1 Luminescent Assay Monitors Fc Effector Activity
Accuracy and precision are critical to the development of bioassays and novel biological agents. Our ADCP Reporter bioassay is both accurate and precise, and has been prequalified in accordance with ICH guidelines, and shows the precision, accuracy and linearity required for routine use in potency and stability studies. We work closely with you to choose the most suitable method based on the immune cells or diseases of interest.
Fig.2 Increasing concentrations of rituximab were incubated with Raji target cells and NFAT-RE effector cells either with or without FcγRIIa-H expression.
Fig.2 Rituximab induces Fc-mediated phagocytosis of Ramos cells by the monocytic cell line THP-1. Engulfment of Ramos cells is confirmed by fluorescence microscopy.
For more detailed information, please feel free to contact us or directly sent us an inquiry.
Creative Biolabs provides luciferase-based ADCC assay. This Jurkat cell based assay is pioneered by Creative Biolabs, and the methodology is very well accepted by the field. See attached ADCC Reporter Assay Protocol for further details.
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Creative Biolabs has established a team of customer support scientists ready to discuss ADCC/CDC optimization strategies, antibody production, bioinformatics analysis and other molecular biology/biotechnology issues.