Development of Monoclonal Antibody PMab-269 Against California Sea Lion Podoplanin
Tomohiro Tanaka, Teizo Asano, Masato Sano, Junko Takei, Hideki Hosono, Ren Nanamiya, Takuro Nakamura, Miyuki Yanaka, Hiroyuki Harada, Masato Fukui, Hiroyoshi Suzuki, Kazuyuki Uchida, Takayuki Nakagawa, Yukinari Kato, Mika K. Kaneko
Abstract
The development of protein-specific antibodies is essential for understanding a wide variety of biological phenomena. Parasitic and viral infections and cancers are known to occur within California sea lion ( Zalophus californianus ) populations. However, sensitive and specific monoclonal antibodies (mAbs) for the pathophysiological analysis of California sea lion tissues have not yet been developed. A type I transmembrane glycoprotein, podoplanin (PDPN), is a known diagnostic marker of lymphatic endothelial cells. We have previously developed several anti-PDPN mAbs in various mammalian species, with applications in flow cytometry, Western blotting, and immunohistochemistry. In this study, we established a novel mAb against California sea lion PDPN (seaPDPN), clone PMab-269 (mouse IgG 1 , kappa), using a Cell-Based Immunization and Screening method. PMab-269 is specifically detected in seaPDPN-overexpressed Chinese hamster ovary (CHO)-K1 cells using flow cytometry and Western blotting. Moreover, PMab-269 clearly identified pulmonary type I alveolar cells, renal podocytes, and colon lymphatic endothelial cells in California sea lion tissues using immunohistochemistry. These findings demonstrate the usefulness of PMab-269 for the pathophysiological analysis of lung, kidney, and lymphatic tissues of the California sea lion.