Editing Monoclonal Antibody Methods (section)

== HER2 ==

Anti-HER2 hybridomas were produced, as described previously.(15) In brief, BALB/c mice were immunized using intraperitoneal (i.p.) injections of 100 mg of recombinant HER2-extracellular domain together with Imject Alum (Thermo Fisher Scientific, Inc.). After several additional immunizations, a booster injection was intraperitoneally administered 2 days before harvesting spleen cells. Spleen cells were then fused with P3U1 cells using PEG1500 (Roche Diagnostics, Indianapolis, IN). The resulting hybridomas were grown in RPMI medium supplemented with hypoxanthine, aminopterin, and thymidine selection medium supplement (Thermo Fisher Scientific, Inc.). Culture supernatants were screened using enzyme-linked immunosorbent assay (ELISA) with recombinant HER2-extracellular domain. MAbs were purified from the supernatants of hybridomas, cultured in Hybridoma-SFM (Thermo Fisher Scientific, Inc.) using Protein G Sepharose 4 Fast Flow (GE Healthcare UK Ltd.).