Humanized Monoclonal Antibody Blocking Carbonic Anhydrase 12 Enzymatic Activity Leads to Reduced Tumor Growth In Vitro

• Published in: Anticancer research Vol. 39; no. 8; pp. 4117 - 4128
• Main Authors: Uda, Narasimha Rao, Stenner, Frank, Seibert, Volker, Herzig, Petra, Markuly, Norbert, VAN Dijk, Marc, Zippelius, Alfred, Renner, Christoph
• Format: Journal Article
• Language: English
• Published: Greece International Institute of Anticancer Research 01.08.2019
• Subjects: Adenocarcinoma; Antibody libraries; Anticancer properties; Antigens; Cancer; Carbonic anhydrase; Carbonic anhydrases; Enzymatic activity; Enzyme-linked immunosorbent assay; Flow cytometry; Lung cancer; Lungs; Monoclonal antibodies; Tumors; Viability; Carbonic anhydrase 12; hypoxia; cancer; therapeutic antibodies
• DOI: 10.21873/anticanres.13570
• ISSN: 0250-7005

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. 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. 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. Our newly identified humanized CA12-antibody with anti-cancer activity, represents a new tool for the treatment of CA12-positive tumors.