Preparation and localization of a monoclonal antibody against a vanadium-associated protein extracted from the blood cells of the vanadium-rich ascidian, Ascidia sydneiensis samea

Ascidians are known to accumulate high levels of vanadium in their blood cells. Recently, we found a vanadium-associated protein (VAP) in blood cells of a vanadium-rich ascidian, Ascidia sydneiensis samea. In this paper, we raised a monoclonal antibody against VAP, designated F8DH. Immunoblot analysis showed that F8DH recognized 2 related peptides of 15 kDa and 16 kDa of VAP. Using F8DH, VAP was shown to be in the cytoplasm of vanadocytes and compartment cells, both of which were reported to contain vanadium. F8DH also stained the vanadocytes distributed in the connective tissues around the alimentary canal, suggesting that vanadocytes in the connective tissue contained VAP. Furthermore, blood cells of 3 different species of ascidian having high levels of vanadium, A. sydneiensis samea, A. ahodori, and Ciona intestinalis, showed reactivity of F8DH but little reactivity was observed in 2 species having less vanadium, Halocynthia roretzi and Pyura michaelseni, suggesting that VAP recognized by F8DH is a common protein in vanadium-rich ascidians.