Preparation and characterization of monoclonal antibodies to eriocheir sinensis hemocyanin and insulin-like growth factor Ⅱ fusion protein

Abstract

The hybridoma lines that secrete anti-ESH monoclonal antibodies were cloned by enzyme-linked immunosorbent assay (ELISA) after fusion of mouse SP2/0-Ag 14 myeloma cells with spleen cells isolated from BALB/c mice that have been immunized with Eriocheir sinensis hemocyanin (ESH), one of arthropod hemocyanin. Anti-ESH monoclonal antibodies were identified to secrete Ig Gl by double gel immunodiffusion. In the case of crossreactivity test with Keyhole limpet hemocyanin (KLH), one of molluscan hemocyanin, anti-ESH monoclonal antibodis and anti-KLH monoclonal antibody showed no affinity each other in ELISA. Insulin-like growth factor II (IGF II) fusion protein was purified by eluton method from SDS-PAGE and the clones that secrete anti-IGF II fusion protein antibodies were selected in HAT medium. And anti-IGF II monoclonal antibodies were identified to secrete Ig Gl by double gel immunodiffusion.