Yağ asiti bağlanma proteini 4(FABP4/aP2)'e karşı monoklonal antikor geliştirilmesi

Abstract

Recombinant proteins and antibodies can be use for therapeutic or diagnostic purposes which produced in many different host organisms. The technique for the production of immortal cell making single antibody, fusing target antibody-forming B lymphocyte precursor with a suitable myeloma cells. The fused hybrid cells (called hybridomas), as a cancer cell will reproduce rapidly and will produce large amounts of the desired antibodies. Fatty acid binding protein (FABP4) is a well characterized intracellular lipid transport protein and plays a key role in the intracellular fatty acid transport and adipose tissue metabolism. FABP4 as a adipokine that regulates glucose homeostasis and has various features for metabolic syndrome associated with obesity. In this study, production of monoclonal antibodies against immunogenic FABP4 protein made by recombinant DNA technology. Recombinant His-FABP4 was expressed in e.coli and purified. Balb/C mice used for immunization and serum anti-FABP4 antibodies determined by enzyme-linked immunosorbent assay (ELISA). Hybridoma cells created by fusion of splenocytes and myeloma partner cells. After selection of antibody producing cell clones, injecting hybridomas into the peritoneal cavity in Balb/C mice ascites fluids was obtained. We have selected fifteen hybridoma clones that produced antibodies specific for FABP4, as shown by western blotting and immunocytochemistry. As a result we produced MAbs that will be useful for the scientific community working on fatty acid binding proteins and lipid metabolism. In near future, therapeutic approach for this antibody maybe a possibility in metabolic syndrome.