Establishment of a Monoclonal Antibody Against chGILZ.

The chicken glucocorticoid-induced leucine zipper (chGILZ) participates in the inflammation of avian immunosuppressive diseases. We aimed to establish a monoclonal antibody (MAb) against chGILZ and to investigate its distribution in clinical diagnosis. A gene cloned from chicken embryo fibroblast (CEF) cell was inserted into the expression vector pET-28a and pGEX-6p-1. The recombinant expression vectors were transformed into Escherichia coli BL21 (DE3). Then the recombinant proteins His-chGILZ and GST-chGILZ were successfully expressed and purified. The hybridomas were developed by fusing mouse myeloma cell line SP2/0 with splenocytes of immunized mice and screened with purified GST-chGILZ. Two hybridomas (1D3 and 3F4) were effective in detecting both recombinant and native chGILZ proteins and were isolated and characterized. The MAbs did not react with human GILZ protein, but could recognize chGILZ by Western blot assay. These data and reagents will be of great assistance to elucidate the molecular mechanism of avian immunosuppressive diseases, such as the infectious bursal disease.