Production And Characterization Of Monoclonal Antibodies Against Siniperca Chuatsi IgM

The purified serum immunoglobulin(Ig) of mandarin fish( Siniperca chuatsi) was used as antigen in preparation of monoclonal antibody (Mab). By using hybridoma-monoclonal antibody technology, eleven hybridoma cell lines which secreting monoclonal antibodies (Mab) against mandarin fish Siniperca chuatsi IgM had been established. And the characteristics of these Mabs had been described. In isotyping analysis, the results showed that among these Mabs, two of them are IgM, three of them are IgG₃, five of them are IgG₁ and one of them belong to IgA; Further experiments proved that Mab7F12-F6 only specifically bound to Ig of siniperca chuatsi; MablF12-A3, 4D3-A7, 3F8-H3, 3A5-D11, 5C3-A7, 5D11-F9, 6G6-A11 are be able to recognise the Ig of Siniperca chuatsi, European eel and Japanese eel. And Mab3D5-H7, 3D3-A6, 4C1-H11 have cross reaction with European eel, Japanese eel and Colossoma brachypomum cuvier. All of them did not have any cross reaction with the Ig of Pseudosciaena crocea, EpinepHelus drummondhayi, Crucian carp, Tilapia, Clarias fuscus, cyprinoid,and also did not react to the fish common bacterial pathogen, such as Aeromonas, Edwardsiella, Vibrio, Salmonella and Escherichia coll. And their ELISA titer ranged from 10⁴ to 10⁶, The sensitivity of the Mabs to purified siniperca chuatsi IgM was at least lower than 39ng. All of these results demonstrated that different fish immunoglobulin have both specifically epitopes of themselves to order and the common epitopes. These common epitopes not only exist between the close relative species, such as the Japanese eel and European eel, but also exist between the distant relative ones such as Siniperca chuatsi, eel and Colossoma brachypomum cuvier. From the deffrent Cross degree between Japanese eel and European eel ,we can also see high likeness exist between Japanese eel serum and European eel serum.But they had specialbility at the same time .We maked the affinity chromatograpHy pillar by using the monoclonal antibody (7F12F6)of mandarin fish linking the Sepharose 4B.Using the way of immunity affinity chromatograpHy to purify serum immunoglobulin(Ig) of...