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Application of Long-term cultured Interferon-γ Enzyme-linked Immunospot Assay for Assessing Effector and Memory T Cell Responses in Cattle
Published on: July 11, 2015
Generation and application of a 293 cell line stably expressing bovine interferon-gamma
Zhengzhong Xu1, Fengli Shan1, Fa Shan1
1Jiangsu Key Lab of Zoonosis/Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China.
Abstract:
A stable mammalian cell line expressing highly active bovine interferon-gamma (BoIFN-γ) was generated using Flp recombinase-mediated integration. This recombinant 293 cell line (B1) efficiently secreted FLAG-tagged BoIFN-γ protein into the culture supernatant, as determined by ELISA and Western blot. The recombinant BoIFN-γ exhibited high anti-viral activity, suggesting that the 293 cells expressed BoIFN-γ that structurally and biologically resembled the natural protein. Two monoclonal antibodies (mAbs) with high affinity for the 293 cell-expressed BoIFN-γ were identified using this cell line, and these mAbs can be used for the development of diagnostic kits. Thus, this work demonstrates the successful generation of a 293 cell line that produces large quantities of highly active BoIFN-γ and demonstrates its potential application in the research of bovine infectious diseases.

