Tolerization of adult mice to immunodominant proteins before monoclonal antibody production

Generating monoclonal antibodies (mAbs) against one polypeptide chain of a heterodimeric protein can be difficult when the other chain is more immunogenic. To influence the immune response in favor of the less immunogenic protein, we rendered adult mice tolerant to the immunodominant protein using a procedure based on the phenomenon of high zone tolerance. We then immunized the tolerized mice with a heterodimeric protein containing the immunogenic protein and produced hybridomas in the usual way. Screening the hybridomas for reactivity against the immunodominant protein and against the heterodimer revealed that this tolerization procedure can result in an increase of hybridomas producing mAbs against the protein of interest by up to 90-fold. This method should be of general utility for the production of mAbs against weakly antigenic proteins in mixtures of antigens.