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Results We use here the CRISPR-Cas 9 based CRISPR-X approach to target AID to antibody genes carried by expression vectors in HEK 293 cells. This directed mutagenesis approach, combined with a highly sensitive antigen-associated magnetic enrichment process, allowed rapid progressive evolution of a human antibody against the Human Leucocyte Antigen A*02:01 allele. Starting from a low affinity monoclonal antibody expressed on Ag-specific naïve blood circulating B cells, we obtained in approximately 6 weeks antibodies with a two log increase in affinity and which retained their specificity.

Ex vivo evolution of human antibodies by CRISPR-X: from a naive B cell repertoire to affinity matured antibodies