Engineering B cells to express fully customizable antibodies with enhanced Fc functions.

Abstract

Genome editing within the constant region of the immunoglobulin Heavy chain locus (IGH) can reprogram B cells to express heavy-chain-only antibodies (HCAbs) containing custom antigen-recognition domains. HCAb-engineered cells express both surface B cell receptor (BCR) and secreted antibody isoforms and respond to antigen. Here, we extend this approach to also allow customization of the constant (Fc) domain of the Heavy chain by selecting alternate editing sites within IGH, producing HCAbs with enhanced effector functions or containing mutations to extend antibody half-life. We also introduced mutations to force obligate HCAb homodimers and prevent unwanted pairing with endogenous antibody chains. Finally, we showed that additional domains could be accommodated at the HCAb C-terminus and preferentially expressed in the secreted isoform. Together, these data demonstrate the flexibility of the HCAb editing platform to express fully customized molecules that take advantage of the properties of B cells.