Fig. 5: Antibody engineered B cells secrete functional linker antibodies.
a Allelic integration frequency in adult peripheral blood CD19+ B cells targeted with AAV6 cassettes for linker antibodies as indicated. Each AAV6 was used at an MOI of 25,000 and integration frequency was measured with ddPCR (dots represent independent biologic donors, n = 8 for mock and 10-1074, n = 5 for Ibalizumab and 10-1074+Ibalizumab, n = 3 for PGDM1400 and CAP256V2LS). Donor 1 (green circle), Donor 2 (blue triangle), and Donor 3 (purple diamond) are specifically noted for their continued use in (b, d, e). Bars represent mean and error bars represent SD. b Concentration of each antibody (10-1074, blue; Ibalizumab, purple; PGDM1400, gold; CAP256V2LS, orange) as determined by antigen specific ELISA from B cell supernatant from cells targeted with AAV6 cassettes as indicated (n = 3 biological donors comprised of Donor 1, Donor 2, and Donor 3 from a). Six days post-editing, B cells were plated at 1 × 106 cells per mL and supernatant was collected after 5 days. Bars represent mean and error bars represent SD. c Measured IC50 for each antibody against TRO11 or CNE55 pseudoviruses as determined by TZM-bl assay in Fig. 1c (NN, not-neutralizing). d Inhibition of infection with TRO11 or e CNE55 HIV-1 pseudovirus by culture supernatant from B cells engineered to express linker antibodies as indicated, as determined by TZM-bl assay. Percent infection for each dose of supernatant from gene targeted B cells is normalized to infection at each dose of supernatant from mock B cells from the same donor (n = 3 biological donors; Donor 1 (green circle), Donor 2 (blue triangle), and Donor 3 (purple diamond) from a). Data points and error bars represent mean with range of technical duplicate infections. Source data are provided as a Source Data file.
