| Author/Editor | | Montanič, Sendi; Terdoslavich, Michela; Rajčevič, uroš; De Leo, Luigina; Bonin, Serena; Čurin-Šerbec, Vladka; Passamonti, Sabina |
| Abstract | | Background. Bilitranslocose (Te 2.A.65.1.1) is a bilirubin-specific membrane transporter, found on absorptive (stomach and intestine) and excretory (kidney and liver) epithelia and in voscular endothelium. Polyclonal antibodies have been raised in rabbits in the post, using a synthetic peptide corresponding to AA 67-75 of rat liver bilitranslocose, as an antigen. Affinity-purified antibodies from immune sera have been found to inhibit various membrane transport functions, including the bilirubin uptake into human hepatocytes and the uptake of some flavonoids into human voscular endothelial cells. It wos described by means of immunohistochemistry using polyclonal antibodies that bilitranslocase expression is severely down-regulated in clear celi renal carcinoma. The aim of our work wos development and characterization of high-affinity, specific mAbs against bilitranslocose, which can be used as a potential diagnostic tool in renal celi carcinoma as well as in a wide variety of biological ossays on different human tissues. Materials and method. Mice were immunized with a multi-antigen peptide corresponding to segment 65-75 of predicted primary structure ()~~,~ bilitranslocase protein. By a sequence of subcloning, immune- and functional test s, we aimed at obtaining a ? monoclonal antibody which recognizes a 37 kDa membrane protein, and influences the transport activity of billjr;~mslocose. Results. On the bosis of previous results, specific IgM monoclonal antibodies were produced in BALB/c mice, in order to further improve and extend the immunological approach to the study of bilitranslocase in renal cancer cells os well as to develop its potential diagnostics use. Conclusions. In this article we show an immunological approach, based on newly developed monoclonal antibodies, to adetailed biochemical and functional characterization of a protein whose gene and protein structure is stili unknown. (Abstract truncated at 2000 characters)
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