Acquirement of HRP conjunct IgG anti-IgMs from most widely cultured freshwater fishes in China and its immunoreactivity

Abstract

Until now, custom-made or commercial polyclonal antibody against only one kind of fi sh IgM limited application of the antibody. During our research on development of vaccine against infection of Clonorchis sinensis (C. sinensis) in several kinds of fi sh, we were conscious of the urgency of secondary antibody to evaluate immune effect and screen C. sinensis infection with immunological technology instead of labor-intensive and time-consuming squash or artifi cial digestion of fi sh fl esh. So that, we purifi ed IgM of grass carp, bighead carp, crucian carp, common carp and tilapia which were widely cultured freshwater fi shes in most areas of China. On this basis, we generated HRP-conjunct rabbit IgG anti-fi sh IgMs with high titers. IgM of other freshwater fi shes including oshima, yellow catfi sh, bream, silver carp and so on could be recognized by the IgG sensitively. Additionally, The ELISA detection displayed that the IgG could be more specifi c and sensitive than custom-made rabbit IgG anti-grass carp IgM. The acquirement of HRP-conjunct rabbit IgG anti-fi sh IgMs was the cornerstone for studying the immune system of teleost fi sh, developing immunoassay methods and evaluation of fi sh vaccine with more convenience.