Cloning and Expression of Brain-Specific Anion Transporter BSAT1 and Isolation of Monoclonal Antibodies to Its Extracellular Fragment

Brain-specifi c anion transporter BSAT1 extracellular fragment (451-557) cDNA was cloned in a vector for prokaryotic expression, a producer E. coli strain was obtained, and recombinant extracellular fragment BSAT1 451-557 was purifi ed and used for immunization of BALB/c mice. Splenic cells from mice with verifi ed immune response were used for hybridoma generation. Several hybridoma clones producing monoclonal antibodies to BSAT1 extracellular fragment were selected. Antibody specifi city was confi rmed by ELISA, immunoblotting with recombinant BSAT1 451-557 , and immunofl uorescent BSAT1 assay on rat brain sections and cultured HEK293 cells. It was demonstrated that the obtained antibodies specifi cally bind native rat and human BSAT1 and can be used in both fundamental studies of structures forming the blood-brain barrier and development of targeted transport of diagnostic preparations and drugs across the blood-brain barrier.