Monoclonal Antibody Core

The Monoclonal Antibody Core produces novel monoclonal antibodies for use in basic research, drug discovery, and clinical applications including diagnosis, surrogate markers for disease status, response to therapy, and drug toxicity. Services include molecular cloning and scale-up of immunization antigens to purification of the resulting antibodies. Researchers can choose to have all of the phases of antibody production conducted by the core or perform particular steps in their own lab.

The core generated a mAb for William Hahn, MD, PhD that recognizes the phosphorylated RalA at residues Ser183/Ser194 by immunizing mice with KLH-conjugated phosphopeptides. These antibodies were used to determine whether Ser183 and Ser194 in RalA were targets of PP2A Aß; a tumor suppressor. The Core designed the peptides, immunized the mice, and conducted primary and secondary screening plus subcloning of the hybridomas.

The antibody the core developed against EGLN2 for William Kaelin, MD, cross-reacted between mouse and human protein. Loss of EGLN2 has been shown to precede loss of Cyclin D, which is overproduced in over 50% of human breast cancers. Our core immunized the mice with recombinant EGLN2 protein, screened against mouse and human recombinant EGLN2 protein, and worked closely with Kaelin's lab to ensure the endogenous protein was also being recognized with the MAb by western blot and immunoprecipitation analysis.