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Introducing dendritic cell antibody internalization as an immunogenicity risk assessment tool.

Rachel MelendezBenjamin OrdoniaJoyce GuerreroAzadeh HassanzadehPeter TranJustin LowManda WongJochen BrummShan ChungLynn Kamen
Published in: Bioanalysis (2022)
Aim: Immunogenicity risk assessment assays are powerful tools that assess the relative immunogenicity of potential biotherapeutics. We detail here the development of a novel assay that measures the degree of antibody internalization by antigen-presenting cells as a predictor of immunogenicity. Results & methodology: The assay uses the fluorescence signal from the antibody bound to the outside of the cell as well as inside the cell to determine internalization. To calculate the amount of internalized antibody, the fluorescent signal from the outside was subtracted from the fluorescent signal from the inside, which is referred to as the internalization index. Conclusion: This assay format demonstrated that antibody-based biotherapeutics with higher clinical immunogenicity internalized to a higher degree than therapeutic antibodies with lower clinical immunogenicity.
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