IN SITU AFFINITY MATURATION OF ANTIBODIES

摘要

This disclosure relates to a method to mimicking the germinal center reaction to generate antibodies with altered binding properties or increased affinity directly in hybridomas. To allow convenient and rapid affinity maturation of antibodies, a controllable activation-induced cytidine deaminase (AID) expression system to induce somatic hypermutation in hybridomas is developed. Selection of high affinity antibodies is achieved by fluorescence-activated cell sorting of hybridomas that preferentially bind fluorescence-labeled antigens. The disclosure also relates to de novo generation of hybridomas with tunable antibody affinity by generating myeloma fusion partners with controllable AID expression.