Narasimha Rao Uda1,2, Frank Stenner3,4, Volker Seibert5,6, Petra Herzig3, Norbert Markuly3,7, Marc VAN Dijk5,8, Alfred Zippelius3,4, Christoph Renner1. 1. Cancer Immunology Laboratory, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland christoph.renner@unibas.ch narasimha.uda@unige.ch. 2. School of Pharmaceutical Sciences, Faculty of Sciences, University of Geneva, Geneva, Switzerland. 3. Cancer Immunology Laboratory, Department of Biomedicine, University Hospital Basel, University of Basel, Basel, Switzerland. 4. Division of Oncology, Department of Internal Medicine, University Hospital Basel, Basel, Switzerland. 5. Agenus Inc. USA (4-Antibody AG), Basel, Switzerland. 6. Lonza AG, Visp, Switzerland. 7. Cureab GmbH, Riehen, Switzerland. 8. AgenTus Therapeutics, Cambridge, U.K.
Abstract
BACKGROUND/AIM: Carbonic anhydrase 12 (CA12) is a membrane-associated enzyme that is highly expressed on many human cancers. It is a poor prognostic marker and hence an attractive target for cancer therapy. This study aimed to develop a humanized CA12-antibody with anti-cancer activity. MATERIALS AND METHODS: Antibody libraries were constructed and screened by the Retrocyte display®. Antibody binding and blocking properties were determined by ELISA, flow cytometry and enzymatic activity assays. Spheroid viability was determined by Cell-Titer-Fluor assay. RESULTS: We developed a novel humanized CA12-specific antibody, 4AG4, which recognized CA12 as an antigen and blocked CA12 enzymatic activity. Our humanized CA12-antibody significantly inhibited spheroid growth of lung adenocarcinoma A549-cells in vitro by blocking CA12 enzymatic activity. Similar anti-tumor effects were recapitulated with CA12-gene knockout of A549-cells. CONCLUSION: Our newly identified humanized CA12-antibody with anti-cancer activity, represents a new tool for the treatment of CA12-positive tumors. Copyright
BACKGROUND/AIM: Carbonic anhydrase 12 (CA12) is a membrane-associated enzyme that is highly expressed on many humancancers. It is a poor prognostic marker and hence an attractive target for cancer therapy. This study aimed to develop a humanized CA12-antibody with anti-cancer activity. MATERIALS AND METHODS: Antibody libraries were constructed and screened by the Retrocyte display®. Antibody binding and blocking properties were determined by ELISA, flow cytometry and enzymatic activity assays. Spheroid viability was determined by Cell-Titer-Fluor assay. RESULTS: We developed a novel humanized CA12-specific antibody, 4AG4, which recognized CA12 as an antigen and blocked CA12 enzymatic activity. Our humanized CA12-antibody significantly inhibited spheroid growth of lung adenocarcinoma A549-cells in vitro by blocking CA12 enzymatic activity. Similar anti-tumor effects were recapitulated with CA12-gene knockout of A549-cells. CONCLUSION: Our newly identified humanized CA12-antibody with anti-cancer activity, represents a new tool for the treatment of CA12-positive tumors. Copyright