Fig. 2. Non-viral precision genome-engineering for clinical-grade cell manufacture.

a, Schematic of the construct design and resulting editing. b, Examples of endogenous TCR knockout and knock-in of up to three neoTCRs in a final clinical-grade cell product. Day 0 (left column) shows an example of the same patient’s enriched T cell product but was not stained with the peptide–HLA multimer. Day 13 flow plots (right 3 columns) show the results of each of the 3 neoTCR product lots for that patient. c, TCR functionality (potency) as evaluated by IFNγ production correlates between small-scale products generated from healthy donor T cells and the final large-scale clinical-grade cell product. The functionality of the neoTCR clinical-grade product made for patient autologous cells (IFNγ EC₅₀ values measured by ELISA or ELLA Simple Plex) was correlated with the functionality of the neoTCR product made in healthy donor cells at product selection (IFNγ EC₅₀ values measured by CBA; Pearson’s r = 0.8412, P < 0.0001). d, Proliferation analysis of the final neoTCR clinical-grade cell product following exposure to peptide–HLA stimulation at 1,000 ng ml^–1. Each dot represents a unique neoTCR product. KO, knockout of the endogenous TCR only, these cells do not have a TCR on their surface; MFI, mean fluorescence intensity; neoTCR⁺ or GE, gene-edited knockout of wild-type TCR and knock-in of neoTCR; WT, wild-type, unedited cells expressing the endogenous TCR.