The immunoglobulin locus of B cells can be reprogrammed by genome editing to produce custom or non-natural antibodies that are not induced by immunization. However, current strategies for antibody reprogramming require complex expression cassettes and do not allow for customization of the constant region of the antibody. Here we show that human B cells can be edited at the immunoglobulin heavy-chain locus to express heavy-chain-only antibodies that support alterations to both the fragment crystallizable domain and the antigen-binding domain, which can be based on both antibody and non-antibody components. Using the envelope protein (Env) from the human immunodeficiency virus as a model antigen, we show that B cells edited to express heavy-chain antibodies to Env support the regulated expression of B cell receptors and antibodies through alternative splicing and that the cells respond to the Env antigen in a tonsil organoid model of immunization. This strategy allows for the reprogramming of human B cells to retain the potential for in vivo amplification while producing molecules with flexibility of composition beyond that of standard antibodies.

B 细胞的免疫球蛋白基因座可以通过基因组编辑进行重新编程，以产生非免疫诱导的定制或非天然抗体。然而，目前的抗体重编程策略需要复杂的表达盒，并且不允许定制抗体的恒定区。在这里，我们证明人类 B 细胞可以在免疫球蛋白重链位点进行编辑，以表达仅重链的抗体，这些抗体支持片段可结晶结构域和抗原结合结构域的改变，这可以基于抗体和非抗体-抗体成分。使用人类免疫缺陷病毒的包膜蛋白 (Env) 作为模型抗原，我们表明，经过编辑以表达针对 Env 的重链抗体的 B 细胞通过选择性剪接支持 B 细胞受体和抗体的调节表达，并且细胞对扁桃体类器官免疫模型中的 Env 抗原。该策略允许对人类 B 细胞进行重编程，以保留体内扩增的潜力，同时产生具有超出标准抗体的组成灵活性的分子。