Title: Preparation and Analysis of Monoclonal Antibodies against Equine Interferon-gamma
Abstract: BALB/c mice(Mus musculus) were immunized intraperitoneally with purified recombinant mEIFN-γ protein. Murine myeloma cells were fused with splenocytes of the immunized mice. An indirect ELISA with recombinant mEIFN-γ derived from baculovirus expression system as antigen was used to screen antibody-producing hybridomas. Twelve hybridomas cells could produce McAbs steadily after 3 cycles of subcloning. Additionally, all McAbs showed positive reaction to mEIFN-γ in indirect fluorescence assay (IFA) tests. Four McAbs were IgG1, two McAbs were IgG2a, three McAbs were IgG2b and three McAbs were IgM isotype, respectively. The light chains of all McAbs were chain. Furthermore, seven McAbs showed positive reaction to GST-mEIFN-γ(1~84) protein, and the others showed positive reaction to GST-mEIFN-γ(80~146) protein in specific ELISA tests. The results indicated that 12 McAbs could recognize two or more mEIFN-γ epitope. Therefore, these McAbs can be useful as a basis of detection methods for equine IFN-γ, monitoring immune state and investigating immunologic mechanism.
Publication Year: 2008
Publication Date: 2008-01-01
Language: en
Type: article
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