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J. Vet. Sci. 2019; 20(1): 51-57  https://doi.org/10.4142/jvs.2019.20.1.51
Novel application of Influenza A virus-inoculated chorioallantoic membrane to characterize a NP-specific monoclonal antibody for immunohistochemistry assaying
Yang-chang Tu1,2, Kuang-yu Chen2, Chung-kung Chen2, Ming-chu Cheng1,3, Shu-hwae Lee1, Ivan-chen Cheng2,*
1Department of Epidemiology, Animal Health Research Institute, Council of Agriculture, New Taipei City 25158, Taiwan
2Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, Taipei 10617, Taiwan
3Department of Veterinary Medicine, College of Veterinary Medicine, National Pingtung University of Science and Technology, Pingtung 91201, Taiwan
Correspondence to: *Tel: +886-2-3366-9912; Fax: +886-2-2366-1475; E-mail: ivancheng@ntu.edu.tw
Received: April 30, 2018; Revised: October 15, 2018; Accepted: November 18, 2018; Published online: January 31, 2019.
Abstract
Monoclonal antibodies (MAbs) are widely applied in disease diagnoses. Herein, we report a MAb, WF-4, against Influenza A virus nucleoprotein (NP), its broad response with Influenza A virus, and its application in an immunohistochemistry (IHC) assay. WF-4 was screened by immunofluorescence assay (IFA). The results showed that its reactivity with baculovirus-expressed full-length recombinant NP (rNP) in Western blot (WB), indicating its IHC applicability. Fifteen Influenza A virus (reference subtypes H1 to H15) infected chicken embryonated chorioallantoic membranes (CAM), fixed by formalin, were all detectable in the WF-4-based IHC assay. Also, the reactivity of the IHC test with NP from experimentally inoculated H6N1 and from all recent outbreaks of H5 subtype avian Influenza A virus (AIV) field cases in Taiwan showed positive results. Our data indicate that CAM, a by-product of Influenza A virus preparation, is helpful for Influenza A virus-specific MAb characterization, and that the WF-4 MAb recognizes conserved and linear epitopes of Influenza A virus NP. Therefore, WF-4 is capable of detecting NP antigens via IHC and may be suitable for developing various tests for diagnosis of Influenza A virus and, especially, AIV infection.
Keywords: Chorioallantoic membrane, Immunohistochemistry, Influenza A virus, Antibodies, Monoclonal, Nucleoproteins


© 2019 The Korean Society of Veterinary Science.

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