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. 2023 Feb 23;15(1):2171248. doi: 10.1080/19420862.2023.2171248

Figure 3.

Assay overview. Cells are taken from a human and seeded in plates, biologics are added, and, thereafter, cells and surrounding media are analyzed. A flowchart of a T cell assay to assess immunogenicity is depicted. In the first step, blood is drawn from a human, and cells in the blood sample are highlighted and taken to the next step. Then cells are seeded in a plate. Different biologics are depicted as IgG antibodies, Fab fragments, Fc fusions, and a bivalent antibody fragment, and added to the cell-containing plate. Next, to the left, an antigen-presenting cell (purple) is depicted with an MHC-molecule displaying a peptide from the candidate biologics to a naïve T cell (bright green) on the right. Below is an activated T cell (dark green) that secretes cytokines. In the last step, multiple T cells are shown at the top, and the sample is divided into two, namely the cells that are phenotyped by flow cytometry to the left; whereas the supernatant, analyzed for cytokines, is illustrated by a plate to the right.

Schematic illustration of a Peripheral Blood Mononuclear Cell (PBMC) immunogenicity assay. A) PBMCs are isolated from healthy donors. B) Isolated cells are cultured in cell media with added candidate biologics. C) Candidate biologics are taken up by antigen presenting cells (APCs) and presented to T cells in culture. If the biologic is immunogenic, this may lead to T cell activation and concurrent cytokine secretion. D) Immunogenicity assessment can then be performed by phenotyping the T cells following stimulation with candidate biologics and measuring cytokine levels in culture supernatant.