Preparation of swine ISG15 polyclonal antibody and its application in classical swine fever virus

  • Xin-Bin Cai
  • Hua-Lin Hou
  • Li-Yuan Zhang
  • Rong-Ze Tang
  • Shan Yin
  • Xiao-Quan Li
  • Ke Zhang
  • Qian-Nan Zhao
  • Xue Ying
  • Li-Juan Su
  • Shi-Kai Sun
  • Xiao-Ning Li
  • Ting Rong Luo
Keywords: Interferon stimulated gene 15 (ISG15), classical swine fever virus (CSFV), polyclonal antibody

Abstract

Interferon-stimulated gene 15 (ISG15) is known to participate in innate and adaptive immune responses against
viral infections. Classical swine fever virus (CSFV) causes a global, economically important and highly contagious
disease in pigs. To explore the interaction between ISG15 and CSFV, the ISG15 gene was amplified, cloned and
expressed in E. coli BL21 cells. The expressed ISG15 protein was purified by Ni-NTA agarose column and used to
prepare a polyclonal mouse antibody. The anti-ISG15 antibody produced in mice could react with ISG15 expressed in
the prokaryotic BL-21 cells as well as natural swine ISG15 expressed in PK-15 cells. This polyclonal antibody against
swine ISG15 was used to perform immunofluorescence assay, western blot and flow cytometry for the detection of the
ISG15 protein and to identify that ISG15 was up-regulated in PK-15 cells after infection with CSFV. This polyclonal
antibody against ISG15 protein is important for further research on the interaction between CSFV and ISG15.

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Published
2020-09-23
How to Cite
Cai, X.-B., Hou, H.-L., Zhang, L.-Y., Tang, R.-Z., Yin, S., Li, X.-Q., Zhang, K., Zhao, Q.-N., Ying, X., Su, L.-J., Sun, S.-K., Li, X.-N., & Luo, T. R. (2020). Preparation of swine ISG15 polyclonal antibody and its application in classical swine fever virus. he hai ournal of eterinary edicine, 50(3), 417-429. etrieved from https://he01.tci-thaijo.org/index.php/tjvm/article/view/245852
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Original Articles