Identify and production of monoclonal antibody of siberian tiger’s immunoglobulin

Abstract:

Abstract:[Objective] Purify Immunoglobulin (Ig) of Siberian Tiger and prepare monoclonal antibody (mAb) against the Ig, they can be used to develop immunological diagnostic kits for diagnosing infectious disease in Siberian Tiger, and also can be used in basic immunity research. [Methods] The Ig of Siberian tigers was purified with saturated ammonium sulfate combined with recombinant Protein G. The C57BL/6 mice were immunized with the purified Ig. Spleno-cytes of the mice immunized were collected and fused with the mouse myeloma cell line (Sp2/0-Ag14). The positive hybridoma clones were selected by ELISA and were identified by western blot. The sandwich ELISA was used to detect immunocompetence of the purified Ig and the mAb. [Results] We obtained three mouse hybridoma clones in this study can produce mAbs against Ig of Siberian Tiger. The derived McAbs can recognize Ig heavy chain of Siberian Tiger specifically. The biological activity of the Ig and obtained McAbs also can be identified by detecting the antibody induced by panleukopenia virus (FPV-HLJ) vaccine in Siberian Tiger. The antibody also would be useful for assess the vaccine effecacy against the infectious disease on the Siberian Tiger. [Conclusion] Protein G can be used in Ig purification of Siberian Tiger. The obtained McAbs from the hybridoma ADT11 in this study owned strong ability to bind Ig of Siberian Tiger and have a stable immunocompetence. They can be used to develop diagnostic methods for detecting infectious disease in Siberian Tiger and vaccine research.